ArticlePDF Available

Modulatory effect of gastric pentadecapeptide BPC 157 on angiogenesis in muscle and tendon healing

Authors:

Abstract and Figures

Angiogenesis is a natural and complex process controlled by angiogenic and angiostatic molecules, with a central role in healing process. One of the most important modulating factors in angiogenesis is the vascular endothelial growth factor (VEGF). Pentadecapeptide BPC 157 promotes healing demonstrating particular angiogenic/angiomodulatory potential. We correlated the angiogenic effect of BPC 157 with VEGF expression using in vitro (cell culture) and in vivo (crushed muscle and transected muscle and tendon) models. Results revealed that there is no direct angiogenic effect of BPC 157 on cell cultures. On the other hand, immunohistochemical analysis of muscle and tendon healing using VEGF, CD34 and FVIII antibodies showed adequately modulated angiogenesis in BPC 157 treated animals, resulting in a more adequate healing. Therefore the angiogenic potential of BPC 157 seems to be closely related to the healing process in vivo with BPC 157 stimulating angiogenesis by up-regulating VEGF expression.
Content may be subject to copyright.
INTRODUCTION
Numerous peptide growth factors are supposed to influence
angiogenesis, and thereby wound healing and tissue
regeneration. The healing process is particularly interesting in
two closely related tissues: muscle and tendon.
Muscle usually heals remarkably well due to highly
developed vasculature network and existence of satellite cells
and muscle derived stem cells. Tendons, which are during
development rich in cells, metabolically active and containing
high number of blood vessels (1), are eventually maturing to
hypocellular, hypovascular and hyponeural structures (2-4).
Tendons do not heal as well as muscle, but the healing process
in both is very complex and under regulation of many different
factors such as vascular endothelial growth factor (VEGF),
fibroblast growth factor (FGF), transforming growth factor beta
(TGF-β), tumor necrosis factor (TNF) and NO (5, 6).
Pentadecapeptide BPC 157 is a peptide effective in muscle
and tendon healing (7-12), applied alone, without any carrier.
In tendon and muscle healing its functional, biomechanical,
and pathohistological beneficial effect is accompanied by
angiogenic action. BPC 157 may directly protect endothelium
(13), influence NO-system, counteract the effect of both NOS-
inhibitor and NO-precursor (14, 15) as well as over expression
of endothelin (16). BPC 157 has a particular wound healing
effect, also in response to vessel injury and coordination of the
expression of multiple genes involved in the pathogenesis of
vascular disease; it stimulates the expression of the early
growth response 1 (EGR-1) gene and expression of the EGR-1
repressor, nerve growth factor 1-A binding protein-2 (nab2)
(17). From these results BPC 157 shows a particular
angiogenic/angiomodulatory potential, worthy of further
investigation.
In this study, we correlated the angiogenic effect of BPC 157
with VEGF expression using in vitroand in vivo models.
MATERIALS AND METHODS
BPC 157
Synthetic pentadecapeptide BPC 157,
GEPPPGKPADDAGLV, M.W. 1419, (Diagen, Ljubljana,
Slovenia) is a part of protein sequence isolated from gastric juice
and after high pressure liquid chromatography purification
theprotein is obtained with 99% purity, having 1-des-Gly peptide
as impurity. It is very stable and applied dissolved in sterile
saline, without any additional carriers.
Cell culture
As an angiogenesis model TCS CellWorks AngioKit (TCS
CellWorks, Buckingham, United Kingdom) was used. Following
manufacturer's instructions we treated cells with pure medium
(TCS CellWorks, Buckingham, United Kingdom), or with
VEGF as positive control (2 ng/ml), Suramin as negative control
(20 uM) and BPC 157 in two different final concentrations (10
ug/ml and 2 ug/ml). Medium was changed on the first, fourth,
JOURNAL OF PHYSIOLOGY AND PHARMACOLOGY 2009, 60, Suppl 7, 191-196
www.jpp.krakow.pl
L. BRCIC1, I. BRCIC2, M. STARESINIC3, T. NOVINSCAK3, P. SIKIRIC3, S. SEIWERTH1
MODULATORY EFFECT OF GASTRIC PENTADECAPEPTIDE BPC 157
ON ANGIOGENESIS IN MUSCLE AND TENDON HEALING
1Institute of Pathology, University of Zagreb Medical School, Zagreb, Croatia; 2Clinical Department of Pathology and Cytology,
University Hospital Center Zagreb, Zagreb, Croatia; 3Department of Pharmacology, University of Zagreb Medical School,
Zagreb, Croatia
Angiogenesis is a natural and complex process controlled by angiogenic and angiostatic molecules, with a central role
in healing process. One of the most important modulating factors in angiogenesis is the vascular endothelial growth
factor (VEGF). Pentadecapeptide BPC 157 promotes healing demonstrating particular angiogenic/angiomodulatory
potential. We correlated the angiogenic effect of BPC 157 with VEGF expression using in vitro (cell culture) and in vivo
(crushed muscle and transected muscle and tendon) models. Results revealed that there is no direct angiogenic effect of
BPC 157 on cell cultures. On the other hand, immunohistochemical analysis of muscle and tendon healing using VEGF,
CD34 and FVIII antibodies showed adequately modulated angiogenesis in BPC 157 treated animals, resulting in a more
adequate healing. Therefore the angiogenic potential of BPC 157 seems to be closely related to the healing process in
vivo with BPC 157 stimulating angiogenesis by up-regulating VEGF expression.
Key words: angiogenesis, pentadecapeptide BPC 157, cell culture, healing, muscle, tendon, vascular endothelial growth factor
seventh and ninth day. Cell cultures were cultivated in usual
conditions on 37°C and 5% CO2. After 11th day cultures were
fixed and tubules were visualized using CD31 antibody (TCS
CellWorks, Buckingham, United Kingdom). Using digital
camera, tubules were transferred into ISSA- special software for
morphometrical analysis (Vamstec, Zagreb, Croatia).
Angiogenic effect was evaluated by counting the number of the
tubule branching on 5 visual fields (microscope objective x 20)
(modification of Jones RA, 18).
Animal models
All experimental protocols were approved by the Ethics
Committee at the University of Zagreb School of Medicine.
Three different, already established and published models were
used (10-12). Animals used were male Wistar Albino rats,
weighting from 280 to 320 g. Six animals per each experimental
group and time interval were used. Animals were anaesthetized
prior to any trauma. Animals were treated either with BPC 157
(10 ug/kg) dissolved in saline, or with equivalent volume of
saline alone (5 ml/kg).
1. Muscle crush injury
Right hind limbs were shaved, and using special system a
force of 0.727 Ns/cm2was delivered to a maximum diameter of
gastrocnemius muscle complex 2 cm proximal to the insertion of
the Achilles tendon (12). The therapy was applied
intraperitoneally immediately after injury and once a day until
one day before sacrificing on 1st, 4th, 7th and 14th day after trauma.
2. Quadriceps muscle transection
The right quadriceps muscle was isolated and transected 1 cm
proximal to patella. The therapy was applied intraperitoneally 30
minutes after injury and once a day until one day before
sacrificing on 4th, 14th, 21st and 28th day after trauma (11).
3. Achilles tendon transaction
Skin incision in the length of 3 cm was performed above the
right Achilles tendon which was transected 0.5 cm proximal to
calcaneus's insertion. After transection, only skin was sutured.
The therapy was applied intraperitoneally 30 min after surgery
and once a day until one day before sacrificing on 1st, 4th, 7th, 10th
and 14th day after transection (10).
Pathohistological analysis
After sacrificing the respective tissue was dissected, fixed
in buffered formalin (pH 7.4) for 24 hours and embedded in
paraffin using standard procedures. For pathohistological
evaluation transversal sections of crushed muscle, longitudinal
section of transected muscle and longitudinal section of
transected tendon were used. Tissue samples were cut
semiserially, stained with hematoxylin and eosin, or
immunohistochemically for FVIII, CD34 and VEGF (Dako,
Glostrup, Denmark) using Dako Autostainer and
manufacturer's protocols. Vascular elements on hematoxylin
and eosin stained slides as well as immunohistocemically
positive elements were examined in a blinded fashion using
hot-spot assessment and ISSA program (V
AMSTEC, Zagreb,
Croatia). From the area of maximal tissue damage detected
using semiserial sections, five high power fields were
randomly selected for analysis.
Stastistical analysis
For analysis of distribution normality in vivo and in vitro
acquired data Kolmogorov-Smirnov test was used. Since no
group had normal distribution Mann-Whitney U test was applied.
Statistical analysis was performed using SPSS 11.5 for Windows
and all values of p<0.05 were considered statistically significant.
RESULTS
Cell culture
The number of branching points of newly formed tubular
structures was without significant difference (p>0.05) between
cell cultures treated with pure medium or with BPC 157 in both
concentrations (10 ng/ml and 2 ng/ml). As expected, VEGF as
positive control had proangiogenic effect, as opposed to negative
control Suramin which practically completely inhibited
branching (Fig. 1).
Animal models
1. Muscle crush
After crush injury (Fig. 2) in control animals angiogenic
response with all tested parameters (number of VEGF, CD34 and
FVIII positive elements (data obtained on hematoxylin-eosin stain
were equal to FVIII presentation and are not presented))
consistently reached its peak at day 7. Increase in CD34 and VEGF
positive elements was obvious since 2 hours, while FVIII showed
192
Fig. 1. The number of branching tubules in cell cultures
demonstrated the absence of positive and negative angiogenic
effect of BPC 157 in in vitroconditions.
M-pure medium; S-Suramin (negative control); V-VEGF
(positive control); B1-BPC 157 10 µg/ml; B2-BPC 157 2 µg/ml;
*- statistically significant difference (p<0.05)
an increase after day 4. On the other hand, angiogenic response
after BPC 157 application showed a shift toward left, implying at
the early interval (2h-4th day) increased blood vessels formation
(VEGF>CD34>FVIII) with gradual decrease in later period (4th -
14th day). VEGF reached its peak the 1st day, as the earliest
breakpoint, while CD34 and FVIII reached their peaks at 4th day.
2. Muscle transection
In concordance with the severity of injury (complete
transection) we should note that angiogenic response was
generally higher in both groups than after crush injury (measured
with the number of VEGF, CD34 and FVIII positive elements),
but without a consistent peak (Fig. 3). Interestingly, curves in
both groups are of the same shape, but with the statistically
significant difference- higher numbers of positive vascular
elements in the BPC 157 treated group. The angiogenic activity
in both groups was obviously prolonged. The highest activity in
this model demonstrated CD34 positive elements with earliest
peak at 4th day, followed by VEGF (peak at 14th day) and FVIII
(peak at 21 day).
3. Transected Achilles tendon
After tendon transection (Fig. 4), in both groups, angiogenic
response demonstrated with all tested parameters (the number of
VEGF, CD34 and FVIII positive elements) consistently reached
its peak already at day 4. In control group the numbers in the first
time periods remained within relatively low values, but toward
the end reaching higher values than BPC 157 animals. The
number of FVIII positive elements showed a second peak only in
control animals, retaining this number of vascular structures till
the end. BPC 157 treated animals showed pronounced decrease
in all parameters after their peak, resulting in low number of
positive vascular elements at the end of investigated period.
DISCUSSION
This study evaluated the angiogenic potential of BPC 157. In
early post-injury periods, BPC 157 therapy induced a prominent
increase of angiogenesis in rats with transected Achilles tendon or
quadriceps muscle and in rats with crushed muscle. This was
consistently visualized with different endothelial cell antigens,
FVIII (involved in platelet adhesion and aggregation, present on
endothelial cells of mature blood vessels) and CD34 (involved in
leukocyte adhesion and endothelial cell migration during
angiogenesis, present on capillary endothelial cells), as well as
with VEGF presentation (main factor in angiogenesis, expressed
on endothelial cells, mitogen for vascular endothelial cells).
Generally, BPC 157 increased the number of VEGF, CD34 and
FVIII positive vascular elements, and angiogenic response was
193
Fig. 2. Results of immunohistochemical
analysis after muscle crush injury,
demonstrating positive angiomodulatory
effect of BPC 157.
Legend: N- number of positive elements,
full line- BPC 157 treated animals;
broken line- controls; time-time after
injury; *- statistically significant
difference (p<0.05).
regularly augmented and shifted toward the left. On the other
hand, results obtained in in vitroconditions, using human
endothelial cells, showed that there is no direct angiogenic effect
of BPC 157 on cell cultures. The angiogenic potential of BPC 157
seems to be closely related to the healing process in vivo with BPC
157 stimulating angiogenesis by up-regulating VEGF expression.
In general, angiogenesis, the formation of new vessels from
pre-existing ones, consists of an orderly sequence of events
triggered by growth factors secreted from the surrounding
hypoxic tissue (19). It is a complex process controlled by
angiogenic and angiostatic molecules resulting in ideal
revascularization of the wound bed, and providing oxygen,
nutrients, and inflammatory cells to the newly
growing/regenerating as well as tumor tissue (20, 21).
Conditions less than ideal (e.g. hypoxia, wound fragments
misadaptation, infection) tend to hamper this process resulting in
scar formation or delayed healing, and probably induce VEGF
formation by more than one mechanism (22). Our study
demonstrated in all investigated models of tendon and muscle
injury that BPC 157 induces higher VEGF and CD34 positivity,
preceding the increase in actual number of blood vessels as
demonstrated on HE and FVIII stains. Such a particular activity
in angiogenesis and healing is concordant with the previous
evidence that BPC 157 may directly protect endothelium (13),
influence NO-system, counteract the effect of NOS-inhibitor and
NO-precursor (14, 15), as well as over expression of endothelin
(16). Beside the stimulation of expression of the EGR-1 gene,
BPC 157 also stimulated expression of nab2 (17). Coordinated
regulation of this transcription factor and its repressor suggests
that this system may play a role in maintaining vascular
homeostasis. It is possible that BPC 157 - nab2 interaction is part
of a feedback mechanism which serves to regulate EGR-1-
mediated gene transcription. The BPC 157 effects on rats with
crushed muscle, transected muscle and tendon suggest
appropriate angiogenic response that results in better healing
(10-12). We assessed the angiogenic response in connection to
different extent of tissue damage (muscle transection vs. muscle
crush) and different tissue healing capacity (muscle vs. tendon).
The detrimental consequence of muscle transection outweighs
crush injury, and after quadriceps muscle transection more
VEGF and CD34 positive elements were present than after blunt
trauma. Consistently, after Achilles tendon transection the same
parameters remained far below the values noted after muscle
transection.
However, after either muscle or tendon had been completely
transected, the commonly negligible tissue repair clearly shows
that without therapy the described angiogenic response may still
be inadequate. In tendon transection model control animals had
two peaks of FVIII positivity, announced by the higher number
of CD34 and VEGF positive elements on the seventh day, in
comparison to the BPC 157 animals. As previously published,
there is no longer visible detachment between Achilles tendon
194
Fig. 3. Results of immunohistochemical
analysis after muscle transection,
demonstrating positive angiomodulatory
effect of BPC 157.
Legend: N- number of positive elements,
full line- BPC 157 treated animals;
broken line- controls; time- time after
transection; *- statistically significant
difference (p<0.05).
ends in BPC animals after seventh day, while it disappears only
after tenth day in control group (10). This space is filled with
granulation tissue with active angiogenic process. Although
angiogenesis is very important for tendon healing, prolonged
angiogenesis may result in prolonged/impaired healing and leads
to chronic tendon disease. Thus, regardless of particularities in
angiogenic response(s), different tissues and different healing
stage, a more generalized and more powerful angiogenic
response induced by BPC 157, with adequately improved
angiogenesis leads consistently to better healing conditions and
thereby eventually to more adequate healing.
It is important to stress that the positive influence of BPC
157 on healing can not be attributed to angiogenesis alone, and
that one should bear in mind its effects on inflammatory
reaction where it decreases leukotriene B4, tromboxan B2 and
myeloperoxidase concentration in injured tissues (23).
Tkalcevic et al. have recently demonstrated a better modulatory
effect on granulation tissue in excisional wounds in genetically
modified diabetic mice db/db in comparison to PDGF-BB
(which is the only approved medication for diabetic ulcers
treatment) (17). Interestingly, PDGF in wounds directly induces
VEGF-A mRNA (24). Tkalcevic et al. also showed that in in
vitroconditions BPC 157 stimulates mRNA EGR-1 (17) having
the same effect as VEGF in cell cultures (25, 26). Interactions
of BPC 157 with the same growth factors and cytokines are
under investigation.
In conclusion, we have demonstrated that, although BPC
157 doesn't have any direct angiogenic effect in cell culture, it
has a positive angiomodulatory effect in animal models of
muscle and tendon healing, resulting in faster and better healing,
which could be helpful in further therapy development.
Conflict of interests: None declared.
REFERENCES
1. Peacock EE Jr. Astudy of the circulation in normal tendons
and healing grafts. Ann Surg1959; 149: 415-428.
2. Ahmed IM, Lagopoulos M, McConnell P, Soames RW,
Sefton GK. Blood supply of the Achilles tendon. J Orthop
Res 1998; 16: 591-596.
3. Gelberman RH. Flexor tendon physiology: tendon nutrition
and cellular activity in injury and repair. Instr Course Lect
1985; 34: 351-360.
4. Schmidt-Rohlfing B, Graf J, Schneider U, Niethard FU. The
blood supply of the Achilles tendon. Int Orthop 1992; 16:
29-31.
5. Hawke TJ, Garry DJ. Myogenic satellite cells: physiology to
molecular biology. J Appl Physiol 2001; 91: 534-551.
6. Stamler JS, Meissner G
. Physiology of nitric oxide in
skeletal muscle. Physiol Rev 2001; 81: 209-237.
195
Fig. 4. Results of immunohistochemical
analysis after Achilles tendon transection,
demonstrating positive angiomodulatory
effect of BPC 157.
Legend: N- number of positive elements,
full line- BPC 157 treated animals;
broken line- controls; time- time after
transection; *- statistically significant
difference (p<0.05).
7. Seiwerth S, Sikiric P, Grebarevic Z, et al. BPC 157's effect
on healing. J Physiol (Paris) 1997; 91: 173-178.
8. Seveljevic-Jaran D, Cuzic S, Dominis-Kramaric M, et al.
Accelerated healing of excisional skin wounds by PL 14736
in alloxan-hyperglycemic rats. Skin Pharmacol Physiol
2006; 19: 266-274.
9. Krivic A, Anic T, Seiwerth S, Huljev D, Sikiric P. Achilles
detachment in rat and stable gastric pentadecapeptide BPC
157: promoted tendon-to-bone healing and opposed
corticosteroid aggravation. J Orthop Res 2006; 24: 982-989.
10. Staresinic M, Sebecic B, Patrlj L, et al. Gastric
pentadecapeptide BPC 157 accelerates healing of transected
rat Achilles tendon and in vitro stimulates tendocytes
growth. J Orthop Res 2003; 21: 976-983.
11. Staresinic M, Petrovic I, Novinscak T, et al. Effective
therapy of transected quadriceps muscle in rat: gastric
pentadecapeptide BPC 157. J Orthop Res 2006; 24: 1109-
1117.
12. Novinscak T, Brcic L, Staresinic M, et al. Gastric
pentadecapeptide BPC 157 as an effective therapy for muscle
crush injury in the rat. Surg Today 2008; 38: 716-725.
13. Sikiric P, Seiwerth S, Grabarevic Z, et al. The beneficial
effect of BPC 157, a 15 amino acid peptide BPC fragment,
on gastric and duodenal lesion induced by restraint stress,
cysteamine and 96% ethanol in rats. A comparative study
with H2 receptor antagonists, dopamine promoters and gut
peptides. Life Sci 1994; 54: PL63-PL68.
14. Grabarevic Z, Tisljar M, Artukovic B, et al. The influence of
BPC 157 on nitric oxide agonist and antagonist induced
lesions in broiler chicks. J Physiol (Paris) 1997; 91: 139-149.
15. Turkovic B, Sikiric P, Seiwerth S, et al. Stable gastric
pentadecapeptide BPC 157 studied for inflammatory bowel
disease (PLD-116, PL14736, Pliva) induces nitric oxide
synthesis. Gastroenterology 2004; 126: 287.
16. Lovric-Bencic M, Sikiric P, Separovic J, et al. Doxorubicine
congestive heart failure-increased big-endothelin 1 plasma
concentration. Reversal by amlodipine, losartan and gastric
pentadecapeptide BPC 157 in rat and mouse. J Pharmacol
Sci 2004; 95: 19-26.
17. Ivetic Tkalcevic V, Cuzic S, Brajsa K, et al. Enhancement by
PL 14736 of granulation and collagen organization in
healing wounds and the potential role of egr-l expression.
Eur J Pharmacol 2007; 570: 212-221.
18. Jones RA, Kotsakis P, Johnson TS, et al. Matrix changes
induced by transglutaminase 2 lead to inhibition of
angiogenesis and tumor growth. Cell Death Differ 2006; 13:
1442-1453.
19. Distler JH, Hirth A, Kurowska-Stolarska M, Gay RE, Gay S,
Distler O. Angiogenic and angiostatic factors in the molecular
control of angiogenesis. QJ Nucl Med 2003; 47: 149-161.
20. Konturek PC, Konturek SJ, Brzozowski T. Helicobacter
pylori infection in gastric cancerogenesis. J Physiol
Pharmacol 2009; 60: 3-21.
21. Ahluwalia A, Li A, Cheng G, Deng X, Tarnawski AS.
Reduced ghrelin in endothelial cells plays important
mechanistic role in aging-related impairment of
angiogenesis. J Physiol Pharmacol 2009; 60: 29-34.
22. Yang HT, Prior BM, Lloyd PG, et al. Training-induced
vascular adaptations to ischemic muscle. J Physiol
Pharmacol 2008; 59: 57-70.
23. Veljaca M, Lesch CA, Pllana R, Sanchez B, Chan K,
Guglietta A. BPC-15 reduces trinitrobenzene sulfonic acid-
induced colonic damage in rats. J Pharmacol Exp Ther
1994; 272: 417 422.
24. Enholm B, Paavonen K, Ristimaki A, et al. Comparison of
VEGF, VEGF-B, VEGF-C and Ang-1 mRNA regulation by
serum, growth factors, oncoproteins and hypoxia. Oncogene
1997; 14: 2475-2483.
25. Kumbrink J, Gerlinger M, Johnson JP. Egr-1 induces the
expression of its corepressor nab2 by activation of the nab2
promoter thereby establishing a negative feedback loop. J
Biol Chem 2005; 280: 42785-42793.
26. Lucerna M, Mechtcheriakova D, Kadl A, et al. NAB2, a
corepressor of EGR-1, inhibits vascular endothelial growth
factor-mediated gene induction and angiogenic responses of
endothelial cells. J Biol Chem 2003; 278: 11433-11440.
R e ce iv e d : October 15, 2009
A c ce pt e d : December 11, 2009
Author's address: Prof. Sven Seiwerth, M.D., Ph. D.,
Institute of Pathology, University of Zagreb Medical School,
Salata 10, 10000 Zagreb, Croatia; Phone: +385 1 4566977;
Fax: +385 1 4921151; E-mail: seiwerth@mef.hr
196
... Likewise, corneal neovascularization might be induced by hypoxic conditions (upregulation of proangiogenic factors, downregulation of anti-anxiogenic factors to supply oxygen to the cornea) [181]. Note, in BPC 157-treated rats, an ameliorated healing course occurred with apparently less aqueous cells [14] and BPC 157 may particularly modulate vascular endothelial growth factor (VEGF) activities [104,192,193] essential for cornea neovascularization. ...
... Thereby, the findings that BPC 157-treated rats generally had no new vessels, and those that did form in the limbus did not make contact with the penetrated area [14,15], which was consistent with evidence. As such, it means essential points are fully resolved for both corneal wound healing and wound healing in general and applied BPC 157 therapy [192,193]. Most importantly, these were in accordance with the identified particular vascular target for successful therapy in the rat glaucoma, retinal ischemia, and corneal ulcers (i.e., episcleral veins, central retinal artery). ...
... Finally, the rapid regaining of corneal transparency illustrates the modulated tissue-specific healing effects of the BPC 157 therapy effects well (cornea vs. other tissues). A strong angiogenic effect was wound healing angiogenesis in the BPC 157 therapy of other tissues [2,6,192,193]. This was evidenced as a part of the ad-vanced healing [2,6,192,193] (note, in hind ischemia recovery, BPC 157 accelerates the blood flow recovery and vessel number) [104], including other avascular tissues [2,6,192,193]; an example of which is tendon healing. ...
Article
Full-text available
Recently, stable gastric pentadecapeptide BPC 157 therapy by activation of collateral pathways counteracted various occlusion/occlusion-like syndromes, vascular, and multiorgan failure, and blood pressure disturbances in rats with permanent major vessel occlusion and similar procedures disabling endothelium function. Thereby, we revealed BPC 157 cytoprotective therapy with strong vascular rescuing capabilities in glaucoma therapy. With these capabilities, BPC 157 therapy can recover glaucomatous rats, normalize intraocular pressure, maintain retinal integrity, recover pupil function, recover retinal ischemia, and corneal injuries (i.e., maintained transparency after complete corneal abrasion, corneal ulceration, and counteracted dry eye after lacrimal gland removal or corneal insensitivity). The most important point is that in glaucomatous rats (three of four episcleral veins cauterized) with high intraocular pressure, all BPC 157 regimens immediately normalized intraocular pressure. BPC 157-treated rats exhibited normal pupil diameter, microscopically well-preserved ganglion cells and optic nerve presentation, normal fundus presentation, nor- mal retinal and choroidal blood vessel presentation, and normal optic nerve presentation. The one episcleral vein rapidly upgraded to accomplish all functions in glaucomatous rats may correspond with occlusion/occlusion-like syndromes of the activated rescuing collateral pathway (azygos vein direct blood flow delivery). Normalized intraocular pressure in glaucomatous rats corresponded to the counteracted intra-cranial (superior sagittal sinus), portal, and caval hypertension, and aortal hypotension in occlusion/occlusion-like syndromes, were all attenuated/eliminated by BPC 157 therapy. Furthermore, given in other eye disturbances (i.e., retinal ischemia), BPC 157 instantly breaks a noxious chain of events, both at an early stage and an already advanced stage. Thus, we further advocate BPC 157 as a therapeutic agent in ocular disease.
... These might represent the general therapeutic effect on the muscle and the healing and functions recovery (and also tendon [58,59,[94][95][96][97], ligament [98] and bone [99][100][101]). These might be the therapeutic effects against those induced by either of the direct injuries [43][44][45][46][47][48]. Likewise, these might be the therapeutic effects against those induced by the various muscle disabilities, and the large variety of different noxious events. ...
... Thereby, the reported myotendinous junction recovery [43] and restoration of full function with the stable gastric pentadecapeptide BPC 157 therapy [43] might have particular therapy potential. This might be taken as the combining healing point of the simultaneously realized recovery of both transected tendon (as well as detached tendon, and osteotendinous junction recovery) [94][95][96][97] and transected muscle [44,48]. In a more extensive way, we should note that a purposive movement requires the impulses passing from the motor cortex via the spinal cord to the appropriate muscles and the movement pattern coordinated by the impulses passing through various parts of the brain and sending messages back to the motor cortex [8]. ...
... Tendon-tendon continuities were reported to have re-established well, with no ossicles forming in other tissues [94][95][96][97][98] (note, with bone morphogenetic proteins (BMPs) [126][127][128], the initial tendon healing process is misleading, due to its similarity to the process of fracture healing [126] and the formation of ossicles in other tissues [126][127][128]). Likewise, with BPC 157 therapy, there was a re-established muscle-muscle continuity, and thereby a re-established tendon-muscle continuity as well [43][44][45][46][47][48][93][94][95][96][97]. Similarly, this might also occur with a ligament transection, with a reestablished ligament-ligament continuity, and fully recovered function upon medial collateral ligament transection [98]. ...
Article
Full-text available
First, we review the definitively severed myotendinous junction and recovery by the cytoprotective stable gastric pentadecapeptide BPC 157 therapy, its healing that might combine both transected and detached tendon and transected muscle, ligament and bone injuries, applied alone, as native peptide therapy, effective in rat injury, given intraperitoneally or in drinking water or topically, at the site of injury. As a follow up, we reviewed that with the BPC 157 therapy, its cytoprotective ability to organize simultaneous healing of different tissues of and full recovery of the myotendinous junction might represent the particular muscle therapy against distinctive etiopathology muscle disabilities and weakness. In this, BPC 157 therapy might recover many of muscle disabilities (i.e., succinylcholine, vascular occlusion, spinal cord compression, stroke, traumatic brain injury, severe electrolyte disturbances, neurotoxins, neuroleptics, alcohol, serotonin syndrome and NO-system blockade and tumor-cachexia). These might provide practical realization of the multimodal muscle-axis impact able to react depending on the condition and the given agent(s) and the symptoms distinctively related to the prime injurious cause symptoms in the wide healing concept, the concept of cytoprotection, in particular. Further, the BPC 157 therapy might be the recovery for the disabled heart functioning, and disabled smooth muscle functioning (various sphincters function recovery). Finally, BPC 157, native and stable in human gastric juice, might be a prototype of anti-ulcer cytoprotective peptide for the muscle therapy with high curing potential (very safe profile (lethal dose not achieved), with suited wide effective range (µg-ng regimens) and ways of application).
... There was also a strong wound-healing effect (for review, see, i.e., [3,102]). Thereby, there was the curing of the skin [53,55,[103][104][105], nerve [106], tendon [50,51,[107][108][109][110][111], muscle [110][111][112][113][114][115], ligament [116], and bone [117][118][119] injuries that spontaneously might not heal. In particular, there was a capability to simultaneously organize the healing of the different tissues (as an example occurred the healing of the osteotendinous junction [108,109] and the healing of the myotendinous junction [111] (and neuromuscular junction function recovering [68]) or the healing of the fistulas, external and internal [120]). ...
... There was also a strong wound-healing effect (for review, see, i.e., [3,102]). Thereby, there was the curing of the skin [53,55,[103][104][105], nerve [106], tendon [50,51,[107][108][109][110][111], muscle [110][111][112][113][114][115], ligament [116], and bone [117][118][119] injuries that spontaneously might not heal. In particular, there was a capability to simultaneously organize the healing of the different tissues (as an example occurred the healing of the osteotendinous junction [108,109] and the healing of the myotendinous junction [111] (and neuromuscular junction function recovering [68]) or the healing of the fistulas, external and internal [120]). ...
... In particular, there was a capability to simultaneously organize the healing of the different tissues (as an example occurred the healing of the osteotendinous junction [108,109] and the healing of the myotendinous junction [111] (and neuromuscular junction function recovering [68]) or the healing of the fistulas, external and internal [120]). Likewise, in particular regard for wounding [3,102], these realized healing effects in the various wounds [53,55,[103][104][105][106][107][108][109][110][111][112][113][114][115][116][117][118][119][120] might evidence the realized healing process after blood vessel are ruptured as a whole, and thereby, as we claimed [59], a distinctive effect on all four major events in clot formation and dissolution was fully accomplished. This meant a highly utilizable special effect, especially with heart failure therapy [18,19,23,24,[27][28][29]31,[37][38][39][40]. ...
Article
Full-text available
In heart disturbances, stable gastric pentadecapeptide BPC 157 especial therapy effects combine the therapy of myocardial infarction, heart failure, pulmonary hypertension arrhythmias, and thrombosis prevention and reversal. The shared therapy effect occurred as part of its even larger cytoprotection (cardioprotection) therapy effect (direct epithelial cell protection; direct endothelium cell protection) that BPC 157 exerts as a novel cytoprotection mediator, which is native and stable in human gastric juice, as well as easily applicable. Accordingly, there is interaction with many molecular pathways, combining maintained endothelium function and maintained thrombocytes function, which counteracted thrombocytopenia in rats that underwent major vessel occlusion and deep vein thrombosis and counteracted thrombosis in all vascular studies; the coagulation pathways were not affected. These appeared as having modulatory effects on NO-system (NO-release, NOS-inhibition, NO-over-stimulation all affected), controlling vasomotor tone and the activation of the Src-Caveolin-1-eNOS pathway and modulatory effects on the prostaglandins system (BPC 157 counteracted NSAIDs toxicity, counteracted bleeding, thrombocytopenia, and in particular, leaky gut syndrome). As an essential novelty noted in the vascular studies, there was the activation of the collateral pathways. This might be the upgrading of the minor vessel to take over the function of the disabled major vessel, competing with and counteracting the Virchow triad circumstances devastatingly present, making possible the recruitment of collateral blood vessels, compensating vessel occlusion and reestablishing the blood flow or bypassing the occluded or ruptured vessel. As a part of the counteraction of the severe vessel and multiorgan failure syndrome, counteracted were the brain, lung, liver, kidney, gastrointestinal lesions, and in particular, the counteraction of the heart arrhythmias and infarction.
... After careful dissecting of quadriceps tendon from the quadriceps muscle, we focused on the myotendinous junction recovery [1,2] and the stable gastric pentadecapeptide BPC 157 therapy [3][4][5][6][7][8][9][10][11][12][13][14][15][16][17] known to heal both transected and detached tendon and transected muscle [18][19][20][21][22], applied alone, as native peptide therapy, that may be effective in rat injury, given intraperitoneally or in drinking water [3][4][5][6][7][8][9][10][11][12][13][14][15][16][17]. ...
... Thereby, for the healed both transected and detached tendon and transected muscle, as native peptide therapy [18][19][20][21][22]27,28], and then the myotendinous junction injury to be healed [1,2], necessitating more advanced strategies for a sustained, safe and reproducible delivery to ascertain the effect, the more general BPC 157 evidence may be also important [3][4][5][6][7][8][9][10][11][12][13][14][15][16][17]. Illustratively, BPC 157 (as an original anti-ulcer peptide, stable in human gastric juice more than 24 h unlike standard growth factors rapidly destroyed, and thereby easy applicable, lethal dose (LD1) not achieved, used to be in ulcerative colitis trials and now multiple sclerosis [3][4][5][6][7][8][9][10][11][12][13][14][15][16][17]), within wound/healing gastrointestinal ulcer relations and angiogenesis is advantageous over standard growth angiogenic factors [7]. ...
... In support, in addition to the healed both transected and detached tendon and transected muscle [18][19][20][21][22], BPC 157 has an especial effect on tendon fibroblasts. ...
Article
Full-text available
(1) Aim: The stable gastric pentadecapeptide BPC 157 is known to heal transected muscle, tendon, and ligament. Thereby, in this study, we investigated the effect of BPC 157 on the dissection of the quadriceps tendon from the quadriceps muscle in rats. (2) Materials and Methods: Myotendinous junction defect, which cannot heal spontaneously in rats, as evidenced with consistent macro/microscopic, biomechanical, functional assessments, eNOS, and COX-2 mRNA levels and oxidative stress and NO-levels in the myotendinous junctions. BPC 157 (10 µg/kg, 10 ng/kg) regimen was given (i) intraperitoneally, first application immediately after surgery, last 24 h before sacrifice; (ii) per-orally, in drinking water (0.16 µg/mL, 0.16 ng/mL, 12 mL/rat/day), till the sacrifice at 7, 14, 28 and 42 postoperative days. (3) Results: These BPC 157 regimens document prominent therapy effects (macro/microscopic, biomechanical, functional much like eNOS and COX-2 mRNA levels and counteracted oxidative stress and NO-levels in the myotendinous junctions), while controls have a poor presentation. Especially, in rats with the disabled myotendinous junction, along with full functional recovery, BPC 157 counteracts muscle atrophy that is regularly progressive and brings muscle presentation close to normal. Accordingly, unlike the perilous course in controls, those rats, when receiving BPC 157 therapy, exhibit a smaller defect, and finally defects completely disappear. Microscopically, there are no more inflammatory infiltrate, well-oriented recovered tissue of musculotendon junction appears in BPC 157 treated rats at the 28 days and 42 days. (4) Conclusions: BPC 157 restores myotendinous junction in accordance with the healing of the transected muscle, tendon, and ligament.
... Namely, as a part of synchronizing healing (19), BPC 157 accelerates the healing of various wounds (i.e. skin (117)(118)(119), muscle (120)(121)(122)(123)(124)(125), tendon (124)(125)(126)(127)(128), ligament (129), bone (130); corneal ulcers (131) and ulcers in the entire gastrointestinal tract (68,(87)(88)(89)(90)(91)(92)(93)(94)(95)(96)(97)(98), along with external and internal fistulas (12)(13)(14)(15)(16)(17)(18) and anastomosis, intestinal perturbations (95)(96)(97)132), nerve (133) and vascular damage (134). Also, once the peritoneum is damaged, the coagulation cascade is set in motion (135); BPC 157 counteracts the whole Virchow triad (51)(52)(53)(54)(55)(56)(57)(58)(59)(60)(61)(62)(63)(64)(65)(66), venous and arterial thrombosis (51)(52)(53)(54)(55)(56)(57)(58)(59)(60)(61)(62)(63)(64)(65)(66)134) and attenuates prolonged bleeding and thrombocytopenia after perforation, amputation, and anticoagulant use (35)(36)(37)(82)(83)(84). ...
... Namely, as a part of synchronizing healing (19), BPC 157 accelerates the healing of various wounds (i.e. skin (117)(118)(119), muscle (120)(121)(122)(123)(124)(125), tendon (124)(125)(126)(127)(128), ligament (129), bone (130); corneal ulcers (131) and ulcers in the entire gastrointestinal tract (68,(87)(88)(89)(90)(91)(92)(93)(94)(95)(96)(97)(98), along with external and internal fistulas (12)(13)(14)(15)(16)(17)(18) and anastomosis, intestinal perturbations (95)(96)(97)132), nerve (133) and vascular damage (134). Also, once the peritoneum is damaged, the coagulation cascade is set in motion (135); BPC 157 counteracts the whole Virchow triad (51)(52)(53)(54)(55)(56)(57)(58)(59)(60)(61)(62)(63)(64)(65)(66), venous and arterial thrombosis (51)(52)(53)(54)(55)(56)(57)(58)(59)(60)(61)(62)(63)(64)(65)(66)134) and attenuates prolonged bleeding and thrombocytopenia after perforation, amputation, and anticoagulant use (35)(36)(37)(82)(83)(84). ...
Article
Using duodenocolic fistula in rats, this study attempts to highlight the particular cytoprotection aspects of the healing of fistulas and therapy potential of the stable gastric pentadecapeptide BPC 157, a cytoprotection mediator (i.e. upgrading minor vessels to induce healing at both fistula's sides). Upon duodenocolic fistula creation (two 'perforated' lesions put together) (assessed at 3, 6, 9, 12, and 15 min), BPC 157, given locally at the fistula, or intragastrically (10 μg/kg, 10 ng/kg), rapidly induces vessel 'recruitment', 'running' toward the defect, simultaneously at duodenum and colon, providing numerous collaterals and branching. The mRNA expression studies done at that time provided strongly elevated (nitric oxide synthase 2) and decreased (cyclooxygenase-2, vascular endothelial growth factor A, nitric oxide synthase (NOS)-1, NOS-3, nuclear factor-kappa-B-activating protein) gene expression. As therapy, rats with duodenocolic fistulas, received BPC 157 10 μg/kg, 10 ng/kg, per-orally, in drinking water till sacrifice, or alternatively, intraperitoneally, first application at 30 min after surgery, last at 24 h before sacrifice, at day 1, 3, 7, 14, 21, and 28. Controls exhibited both defects persisting, continuous fistula leakage, diarrhea, continuous weight loss, advanced adhesion formation and intestinal obstruction. Contrary, all BPC 157-treated rats have closed both defects, duodenal and colonic, no fistula leakage (finally, maximal instilled volume corresponds to healthy rats), no cachexia, the same weight as before surgery, no diarrhea, markedly less adhesion formation and intestinal passage obstruction. Thus, BPC 157 regimens resolve the duodenal/colon lesions and duodenocolic fistulas in rats, and rapid vessels recovery appears as the essential point in the implementation of the cytoprotection concept in the fistula therapy.
... Commonly, these may be the general terms of the realization of the brain-gut axis functioning [2,6]. With the BPC 157 therapy the described improved purposive movement rationale (via the motor cortex -spinal cord -appropriate muscles and vice versa) might conceptualize in the brain-muscle axis and muscle-brain axis bidirectional functioning the healing and function recovery of the myotendinous junction (dissection) [50], the muscle lesion (transection, contusion, and corticosteroid application), and nerve (transection) [50,[150][151][152][153][154]. With muscle weakness, these might perceive both prime (i.e. ...
... [50, [150][151][152][153][154] The described improved purposive movement rationale (via the motor cortex-spinal cord-appropriate muscles and vice versa) might conceptualize in the brain-muscle axis function, the healing and function recovery of the myotendinous junction (dissection), the muscle lesion (transection, contusion, and corticosteroid application), and the nerves (transection). [68] With counteracted muscle weakness, stroke was counteracted. ...
Article
Full-text available
Conceptually, a wide beneficial effect, both peripherally and centrally, might have been essential for the harmony of brain–gut and gut–brain axes’ function. Seen from the original viewpoint of the gut peptides’ significance and brain relation, the favorable stable gastric pentadecapeptide BPC 157 evidence in the brain–gut and gut–brain axes' function might have been presented as a particular interconnected network. These were the behavioral findings (interaction with main systems, anxiolytic, anticonvulsive, antidepressant effect, counteracted catalepsy, and positive and negative schizophrenia symptoms models). Muscle healing and function recovery appeared as the therapeutic effects of BPC 157 on the various muscle disabilities of a multitude of causes, both peripheral and central. Heart failure was counteracted (including arrhythmias and thrombosis), and smooth muscle function recovered. These existed as a multimodal muscle axis impact on muscle function and healing as a function of the brain–gut axis and gut–brain axis as whole. Finally, encephalopathies, acting simultaneously in both the periphery and central nervous system, BPC 157 counteracted stomach and liver lesions and various encephalopathies in NSAIDs and insulin rats. BPC 157 therapy by rapidly activated collateral pathways counteracted the vascular and multiorgan failure concomitant to major vessel occlusion and, similar to noxious procedures, reversed initiated multicausal noxious circuit of the occlusion/occlusion-like syndrome. Severe intracranial (superior sagittal sinus) hypertension, portal and caval hypertensions, and aortal hypotension were attenuated/eliminated. Counteracted were the severe lesions in the brain, lungs, liver, kidney, and gastrointestinal tract. In particular, progressing thrombosis, both peripherally and centrally, and heart arrhythmias and infarction that would consistently occur were fully counteracted and/or almost annihilated. To conclude, we suggest further BPC 157 therapy applications.
... Note, BPC 157 therapy can attenuate/eliminate both progressing thrombosis and hemorrhage in occlusion/occlusion-like syndromes [40][41][42][43][47][48][49][50][51][52][53][54][55][56] and attenuate bleeding following amputation [128][129][130], organ perforation [49,173,174], anticoagulants [128][129][130], and antiplatelet agent [128][129][130] application, and resolved even advanced Virchow triad circumstances [40][41][42][43][47][48][49][50][51][52][53][54][55][56]. The increased expressions of VEGF, CD34, and FVIII were shifted toward the left in crushed muscle, transected muscle, and transected tendon [288] along with improved angiogenesis along with improved healing, as particular effect (note, BPC 157 cures corneal ulcer and maintains corneal transparency [113], and inhibits VEGF effect in human melanoma cell line [289]). This can be performed by controlling the VGEF system as well. ...
Article
Full-text available
We highlight the particular aspects of the stable gastric pentadecapeptide BPC 157 pleiotropic beneficial activity (not destroyed in human gastric juice, native and stable in human gastric juice, as a cytoprotection mediator holds a response specifically related to preventing or recovering damage as such) and its possible relations with neurotransmitter activity. We attempt to resolve the shortage of the pleiotropic beneficial effects of BPC 157, given the general standard neurotransmitter criteria, in classic terms. We substitute the lack of direct conclusive evidence (i.e., production within the neuron or present in it as a precursor molecule, released eliciting a response on the receptor on the target cells on neurons and being removed from the site of action once its signaling role is complete). This can be a network of interconnected evidence, previously envisaged in the implementation of the cytoprotection effects, consistent beneficial particular evidence that BPC 157 therapy counteracts dopamine, serotonin, glutamate, GABA, adrenalin/noradrenalin, acetylcholine, and NO-system disturbances. This specifically includes counteraction of those disturbances related to their receptors, both blockade and over-activity, destruction, depletion, tolerance, sensitization, and channel disturbances counteraction. Likewise, BPC 157 activates particular receptors (i.e., VGEF and growth hormone). Furthermore, close BPC 157/NO-system relations with the gasotransmitters crossing the cell membrane and acting directly on molecules inside the cell may envisage particular interactions with receptors on the plasma membrane of their target cells. Finally, there is nerve-muscle relation in various muscle disturbance counteractions, and nerve-nerve relation in various encephalopathies counteraction, which is also exemplified specifically by the BPC 157 therapy application.
Article
Full-text available
Since the early 1990s, when Robert’s and Szabo’s cytoprotection concept had already been more than one decade old, but still not implemented in therapy, we suggest the stable gastric pentadecapeptide BPC 157 as the most relevant mediator of the cytoprotection concept. Consequently, it can translate stomach and gastrointestinal mucosal maintenance, epithelium, and endothelium cell protection to the therapy of other tissue healing (organoprotection), easily applicable, as native and stable in human gastric juice for more than 24 h. These overwhelm current clinical evidence (i.e., ulcerative colitis, phase II, no side effects, and no lethal dose (LD1) in toxicology studies), as BPC 157 therapy effectively combined various tissue healing and lesions counteraction. BPC 157 cytoprotection relevance and vascular recovery, activation of collateral pathways, membrane stabilizer, eye therapy, wound healing capability, brain–gut and gut–brain functioning, tumor cachexia counteraction, muscle, tendon, ligament, and bone disturbances counteraction, and the heart disturbances, myocardial infarction, heart failure, pulmonary hypertension, arrhythmias, and thrombosis counteraction appeared in the recent reviews. Here, as concept resolution, we review the counteraction of advanced Virchow triad circumstances by activation of the collateral rescuing pathways, depending on injury, activated azygos vein direct blood flow delivery, to counteract occlusion/occlusion-like syndromes starting with the context of alcohol-stomach lesions. Counteraction of major vessel failure (congested inferior caval vein and superior mesenteric vein, collapsed azygos vein, collapsed abdominal aorta) includes counteraction of the brain (intracerebral and intraventricular hemorrhage), heart (congestion, severe arrhythmias), lung (hemorrhage), and congestion and lesions in the liver, kidney, and gastrointestinal tract, intracranial (superior sagittal sinus), portal and caval hypertension, aortal hypotension, and thrombosis, peripherally and centrally.
Preprint
Full-text available
Sažetak Istražen je učinak pentadekapeptida BPC 157 na tovnost muških tovnih pilića. Po prvi puta in vivo primijenjen je jednokratno netom izleženim pilićima i to postupkom nebulizacije ultrazvučnim raspršivaćem SONOVAC ® , aerosolom veličine čestica 3-5 mikrona. Svako je pile udahnulo približno količinu 7,5 µg BPC 157 tijekom 60 odnosno upola manje tijekom 30 sekundi. Učinak se je u oba slučaja očitovao značajno bržim prirastom tjelesne mase u odnosu na kontrolnu skupinu kojoj je istim postupkom dana fiziološka otopina PBS. Nakon 35 dana tova pilići skupine kojoj je BPC 157 primijenjen tijekom 30 sekundi bili su najteži (2297,0 g) uz istodobno najmanji utrošak krmne smjese za ostvareni prirast (1,64 kg/kg). Najveću tjelesnu masu postigli su pilići u dobi 42 dana (3032,28 g) ali uz veći utrošak krme u odnosu na kontrolnu skupinu (1,73 prema 1,54 kg/kg). BPC 157 primijenjen je po prvi puta netom izleženim muškim pilićima uz postizanje veće tjelesne mase u odnosu na kontrolnu skupinu. Također dokazano je da je učinak BPC 157 ovisan o primijenjnoj količini. Postignute rezultate treba prihvatiti kao osnovu za nastavak istraživanja. Abstract The effect of pentadecapeptide BPC 157 on male fattening chickens was investigated. For the first time in vivo it was administered once to a net hatched chicken by an ultrasonic spray nebulization process SONOVAC®, a particle size aerosol of 3-5 microns. Each saw inhaled approximately 7.5 µg of BPC 157 over 60 and half less over 30 seconds, respectively. The effect in both cases manifested a significantly faster body mass control in relation to the same saline. After 35 days of fattening, the chickens of the group to which BPC 157 was applied for 30 seconds were the heaviest (2297, g) at the same time the lowest consumption of compound feed for increment (1.64 kg/kg). The largest body weight was achieved by chickens aged 42 days (3032.28 g) but with a higher feed consumption. BPC 157 was administered for the first time to net hatched male chickens with a higher body weight compared to the control group. It has also been proven that the effect of BPC 157 depends on the amount applied. The results achieved should be accepted as a basis for continuing the research.
Article
Full-text available
Ghrelin, a hormone produced mainly by gastric mucosal cells stimulates growth hormone (GH) release. Ghrelin is also expressed in the endothelial cells of blood vessels suggesting its physiological role and a function in these cells. We recently demonstrated that ghrelin induces angiogenesis--new capillary blood vessel formation- in neonatal human microvascular endothelial cells (HMVECs). Angiogenesis is impaired in aging individuals both in vitro and in vivo, but the precise mechanism(s) of this phenomenon is unknown. We examined whether HMVECs derived from aging individuals (66 years and 90 years old), 66-HMVECs and 90-HMVECs have reduced ghrelin levels vs. neonatal (Neo) HMVECs and whether treatment with exogenous ghrelin can restore impaired in vitro angiogenesis on matrigel in aged HMVECs. Ghrelin levels were reduced in the aged HMVECs by 3.2-fold (p<0.05) compared to Neo-HMVECs. Angiogenesis was significantly decreased in the aged 66- and 90-HMVECs by 39.7% (p = 0.003) and 62.4% (p = 0.003), respectively compared to Neo-HMVECs. Treatment with exogenous ghrelin significantly reversed impaired angiogenesis in aged HMVECs with the EC(50) 0.05 nM. Ghrelin induced angiogenesis in Neo-HMVECs mainly through ERK2 activation. This study is the first demonstration that reduced ghrelin is one of the factors responsible for aging-related impairment of angiogenesis.
Article
Full-text available
Stable gastric pentadecapeptide BPC 157 accelerates the healing of a transected Achilles tendon and a transected quadriceps muscle. It may also be of clinical relevance as a systemic and local peptide treatment for crush injury of a major muscle, such as gastrocnemius muscle complex. BPC 157 is effective without a carrier, and it is presently undergoing trials for inflammatory bowel disease, and no toxicity has so far been reported. In crushed rats (force delivered 0.727 Ns/cm2), BPC 157 was applied either intraperitoneally or locally, as a thin cream layer, immediately after injury (sacrifice at 2 h), and once a day for 14 days. BPC 157 improved muscle healing, macroscopically (less hematoma and edema, no post-injury leg contracture), microscopically, functionally, and also based on enzyme activity (creatine kinase, lactate dehydrogenase, aspartate aminotransferase, alanine aminotransferase). BPC 157, at all investigated intervals, given locally or intraperitoneally, accelerated post-injury muscle healing and also helped to restore the full function.
Article
Full-text available
The vascular endothelial growth factor (VEGF) family has recently been expanded by the isolation of two additional growth factors, VEGF-B and VEGF-C. Here we compare the regulation of steady-state levels of VEGF, VEGF-B and VEGF-C mRNAs in cultured cells by a variety of stimuli implicated in angiogenesis and endothelial cell physiology. Hypoxia, Ras oncoprotein and mutant p53 tumor suppressor, which are potent inducers of VEGF mRNA did not increase VEGF-B or VEGF-C mRNA levels. Serum and its component growth factors, platelet-derived growth factor (PDGF) and epidermal growth factor (EGF) as well as transforming growth factor-beta (TGF-beta) and the tumor promoter phorbol myristate 12,13-acetate (PMA) stimulated VEGF-C, but not VEGF-B mRNA expression. Interestingly, these growth factors and hypoxia simultaneously downregulated the mRNA of another endothelial cell specific ligand, angiopoietin-1. Serum induction of VEGF-C mRNA occurred independently of protein synthesis; with an increase of the mRNA half-life from 3.5 h to 5.5-6 h, whereas VEGF-B mRNA was very stable (T 1/2>8 h). Our results reveal that the three VEGF genes are regulated in a strikingly different manner, suggesting that they serve distinct, although perhaps overlapping functions in vivo.
Article
The protection of stomach and duodenum in conjecture with anti-inflammatory effect was demonstrated for a novel 15 amino acid peptide, coded BPC 157, a fragment of the recently discovered gastric juice peptide BPC. BPC 157 (i.p./i.g.) was investigated in rats in comparison with several reference standards in three experimental ulcer models (48 h-restriant stress, subcutaneous cysteamine, intragastrical 96% ethanol ulcer tests) (pre-/co-/post-treatment). Only BPC 157 regimes were consistently effective in all of the tested models. On the other hand, bromocriptine, amantadine, famotidine, cimetidine and somatostatin were in effective (restraint stress). A dose-dependent protection (cysteamine) and/or partial positive effect (related to treatment conditions) (ethanol), was obtained with glucagon, NPY and secretin whereas CCK/26– 30/ was not effective. Based on Monastral blue studies BPC 157 beneficial effect appears to be related to a strong endothelial protection.
Article
Gastric cancer (GS) remains one of the most common cancers worldwide. It is considered as the second most frequent cause of cancer death worldwide, although much geographical variation in incidence exists. Many studies before linked Helicobacter pylori (Hp) which is now considered as an important pathogen, to the risk of developing noncardia GS. This overview attempts to summarize the recent basic and clinical evidence on the link between H. pylori and gastric cancer, after the award of the Nobel Prize for Physiology or Medicine to Drs. J.R. Warren and B.J. Marshall for the first culture and isolation of Hp and the investigation of their relevance to peptic ulcer disease. It become evident that Hp eradication by antibiotic treatment combined with proton pump inhibitor (PPI) serves as the primary chemoprevention strategy to reduce gastric cancer incidence. Moreover, the eradication therapy reduces gastric cancer incidence in patients without any precancerous lesions at the baseline and is most effective before the development of atrophic gastritis. Due to understanding the molecular nature of GC which has been nowadays under intense investigation, our review attempts to highlight recent progress in the field of research on Hp-induced GS. We discuss the geographical diversity in Hp infection and cancer incidence and the mechanistic role of gastrin, cyclooxygenase-2 (COX-2), growth factor, nitric oxide (NO)/NO synthase and E-cadherin/beta-cathenin systems, apoptosis and angiogenesis in Hp-induced gastric carcinogenesis. In addition host-related genetic susceptibility and the role of overexpression of a proinflammatory cytokines and their polymorphism is discussed in the relation to the cascade of events such as gastric atrophy, intestinal metaplasia and dysplasia that finally lead to adenocarcinoma.
Article
Peripheral arterial insufficiency is a progressive degenerative disease associated with an increased morbidity and mortality. It decreases exercise tolerance and often presents with symptoms of intermittent claudication. Enhanced physical activity is one of the most effective means of improving the life of affected patients. While this occurs for a variety of reasons, vascular remodeling can be an important means for improved oxygen exchange and blood flow delivery. Relevant exercise-induced signals stimulate angiogenesis, within the active muscle (e.g. hypoxia), and arteriogenesis (enlargement of pre-existing vessels via increased shear stress) to increase oxygen exchange and blood flow capacity, respectively. Evidence from pre-clinical studies shows that the increase in collateral blood flow observed with exercise progresses over time of training, is accompanied by significant enlargement of isolated collateral vessels, and enhances the responses observed with angiogenic growth factors (e.g. VEGF, FGF-2). Thus, enhanced physical activity can be an effective means of enlarging the structure and function of the collateral circuit. Interestingly, disrupting normal NO production (via L-NAME) eliminates this increase in collateral blood flow induced by training, but does not disturb the increase in muscle capillarity within the active muscle. Similarly, inhibiting VEGF receptor kinase activity eliminates the increase in collateral-dependent blood flow, and lessens, but does not eliminate, angiogenesis within the calf muscle. These findings illustrate distinctions between the processes influencing angiogenesis and arteriogenesis. Further, sympathetic modulation of the collateral circuit does not eliminate the increase in collateral circuit conductance induced by exercise training. These findings indicate that structural enlargement of the collateral vessels is essential to realize the increase in collateral-dependent blood flow capacity caused by exercise training. This raises the potential that meaningful vascular remodeling can occur in patients with intermittent claudication who actively participate in exercise training.
Article
We have used a new technique of epoxy resin injection to examine the blood supply of the Achilles tendon. The posterior distal part showed poor vascularisation, as did the middle part of the tendon. Ruptures occur in the middle part, but only rarely in the distal part. We therefore suggest that there is no direct relationship between blood supply and the frequency of rupture.
Article
Scientific studies of the past 20 years have done much to redefine the mechanisms by which flexor tendons heal. Several points have become increasingly clear: Flexor tendons are nourished to a greater extent by synovial fluid diffusion than vascular perfusion. Tendon cells are capable of proliferating, producing collagen, and reconstructing their own gliding surface in the absence of adhesion ingrowth. The key to a successful outcome after flexor tendon repair appears to be an early restoration of tendon continuity, reconstruction of the sheath, if possible, and early passive mobilization. This complex stimulates the tendon's intrinsic repair potential, which is contained within the cells of the tendon itself but appears to be expressed only under ideal experimental and clinical situations.
Article
The effect of BPC-15 (Booly Protection Compound-15) was evaluated in a rat model of colonic injury. A single intracolonic administration of trinitrobenzene sulfonic acid (TNBS) dissolved in ethanol induces severe colonic damage, which is characterized by areas of necrosis surrounded by areas of acute inflammation. The damage is associated with high myeloperoxidase (MPO) activity, mainly as a reflection of neutrophilic infiltration into the damaged tissue. In this study, 1 hr before a single intracolonic administration of 50 mg/kg of TNBS in 50% ethanol, the animals were treated with one of the following doses of BPC-15: 0.0001, 0.001, 0.01, 0.1, 1 or 10 nmol/kg administered i.p. or with a dose of 10 nmol/kg administered intracolonically. The animals were sacrificed 3 days later and the extent of colonic necrosis and hyperemia was measured with an image analyzer. The i.p. administration of BPC-15 significantly reduced the extent of TNBS-induced colonic damage in a dose-dependent manner. This was associated with a statistically significant and dose-dependent reduction in colonic tissue MPO activity. At the dose tested (10 nmol/kg), intracolonic administration of BPC-15 did not significantly reduce either the extent of the colonic damage or the increase in MPO activity induced by TNBS. In conclusion, this study showed that i.p. administration of BPC-15 reduced TNBS-induced colonic damage in rats.
Article
The 15 amino acid agent BPC 157, showing a wide range of organoprotective action in different experimental models, was used in our experiments in order to establish its influence on different elements connected with the healing process. Elements thought to be of greatest importance in the process of healing are formation of granulation tissue, angiogenesis and production of collagen. In our work we tested the influence of BPC 157 on: granulation tissue and collagen formation, on angiogenesis as well as on tensile strength development, using three experimental rat models: 1) skin incisional wounds; 2) colon-colon anastomoses; and 3) angiogenesis model with synthetic sponge implantation. The specimens were histologically assessed for collagen, reticulin and blood vessels using scoring and morphometry. In all experiments significant differences between BPC 157-treated animals and controls were found, showing a strong, promoting involvement of BPC in the healing process. It is worth noting that these effects were achieved by different routes of application, including intragastric and local, making BPC 157 a potentially useful therapeutic agent.