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Beneficial effects of Aloe vera in treatment of diabetes: Comparative in vivo and in vitro studies

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Amira Abo-youssef at Beni-Suef University
Amira Abo-youssef
Basim Messiha at Beni-Suef University
Basim Messiha
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Abstract and Figures

In the present investigation, the antidiabetic effect of Aloe vera leaf pulp extract was studied in vivo and in vitro as compared to glimiperide. Diabetes was induced experimentally in adult male albino rats by single-dose intraperitoneal injection of streptozotocin (50 mg/kg body weight). The in vitro study was performed using isolated islets of pancreas from adult female albino rats. Both aloe extract (10 ml/kg, p.o.) and glimiperide (10 mg/kg, p.o.) significantly decreased serum glucose and significantly increased serum insulin levels as compared to control diabetic rats. Serum levels of malondialdehyde (MDA) and superoxide dismutase (SOD) were significantly decreased while blood glutathione (GSH) was significantly increased by aloe treatment as compared to diabetic rats. Effect of aloe was better than the effect of glimiperide. Regarding the in vitro study, both aloe (10 μl/l) and glimiperide (10 μmol/l) significantly increased both basal and stimulated insulin secretion from the isolated islets of pancreas as compared to control. These results show a promising antidiabetic effect of aloe for further clinical trials regarding clinical use of aloe extract for treating type II diabetes.
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ORIGINAL ARTICLE
Beneficial effects of Aloe vera in treatment of
diabetes: Comparative in vivo and in vitro studies
Amira Mourad Hussein Abo-Youssef
*
, Basim Anwar Shehata Messiha
Department of Pharmacology and Toxicology, Faculty of Pharmacy, Beni-Sueif University, Egypt
Received 4 December 2011; accepted 31 March 2012
Available online 10 January 2013
KEYWORDS
Aloe;
Diabetes;
Glimiperide;
Rats
Abstract In the present investigation, the antidiabetic effect of Aloe vera leaf pulp extract was stud-
ied in vivo and in vitro as compared to glimiperide. Diabetes was induced experimentally in adult male
albino rats by single-dose intraperitoneal injection of streptozotocin (50 mg/kg body weight). The
in vitro study was performed using isolated islets of pancreas from adult female albino rats.
Both aloe extract (10 ml/kg, p.o.) and glimiperide (10 mg/kg, p.o.) significantly decreased serum glu-
cose and significantly increased serum insulin levels as compared to control diabetic rats. Serum levels
of malondialdehyde (MDA) and superoxide dismutase (SOD) were significantly decreased while
blood glutathione (GSH) was significantly increased by aloe treatment as compared to diabetic rats.
Effect of aloe was better than the effect of glimiperide. Regarding the in vitro study, both aloe (10 ll/l)
and glimiperide (10 lmol/l) significantly increased both basal and stimulated insulin secretion from
the isolated islets of pancreas as compared to control. These results show a promising antidiabetic
effect of aloe for further clinical trials regarding clinical use of aloe extract for treating type II diabe-
tes.
ª 2012 Faculty of Pharmacy, Cairo University. Production and hosting by Elsevier B.V. All rights
reserved.
1. Introduction
Diabetes mellitus is a serious chronic metabolic disorder char-
acterized by a decrease or cessation of insulin secretion in re-
sponse to normal physiological stimuli, or reduced
responsiveness of peripheral tissues to insulin.
1
The metabolic
abnormalities of diabetes cause mitochondrial superoxide
overproduction in endothelial cells of both large and small ves-
sels, as well as in the myocardium. Therefore, oxidative stress
plays a major role in the development of diabetes complica-
tions, both microvascular and cardiovascular complications.
2,3
Many authors claimed for antidiabetic and antioxidant po-
tential of aloe in experimentally-induced diabetes.
4,5
The glu-
cose lowering effects of Aloe vera were suggested to be
mediated, at least partially, through its potent anti-oxidant ef-
fect.
6,7
Recent approaches suggest that treatment of diabetes
should not only focus on insulin secretion but also on antiox-
idant protection of the b-cell. This may facilitate the repair of
b-cells undergoing damage by oxidative stress secondary to
hyperglycemia.
8
*
Corresponding author. Address: 14 Bashar Ibn Bord St.Nasr City,
Cairo, Egypt. Tel.: +20 022748371, mobile: +0122 24 24 884.
E-mail address: dramira73@hotmail.com (A.M.H. Abo-Youssef).
Peer review under responsibility of Faculty of Pharmacy, Cairo
University
Production and hosting by Elsevier
Bulletin of Faculty of Pharmacy, Cairo University (2013) 51, 7–11
Cairo University
Bulletin of Faculty of Pharmacy, Cairo University
www.elsevier.com/locate/bfopcu
www.sciencedirect.com
1110-0931 ª 2012 Faculty of Pharmacy, Cairo University. Production and hosting by Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.bfopcu.2012.03.002
Based on these facts, this research is designed to correlate
antioxidant and antidiabetic effects of aloe in experimentally-
induced diabetes in vivo. The results will also be supported
by an in vitro study on isolated pancreatic islets.
2. Materials and Methods
2.1. Animals
Adult male albino rats of Wistar strain weighing about 200–
250 g were used for the in vivo experiments. Adult female albino
rats of Wistar strain weighing 150–200 g were used for the
in vitro experiments. Animals were obtained from the National
Research Center, Cairo, Egypt. The animals were housed in
plastic cages (28 cm · 43 cm · 18 cm) and were maintained un-
der conventional laboratory conditions throughout the study.
They were fed standard pellet chow (El-Nasr Chemical Co.,
Cairo, Egypt) and were allowed water ad libitum. Animals de-
scribed as fasted had been deprived of food for 18 h but had been
allowed free access to water. This design is accepted by the ethics
committee in the Faculty of Pharmacy Beni Sueif University.
2.2. Preparation of A. vera leaf pulp extract
A. vera leaves, over 3 years old, were washed, weighed, peeled
and the leaf pulp (gel together with latex) was scratched with a
spoon.
5
The pulp was homogenized with a homogenizer (Ul-
tra-Turrax T25, IKA Labortechnik, Germany), mixed with
an equal volume of phosphate buffered saline (0.1 M,
pH = 7), homogenized again, kept at 4 C overnight then fil-
tered through cloth. The clear filtrate was kept at 20 Cin
small portions until use. The yield of fresh aloe pulp was about
35% v/w in terms of starting fresh leaf weight.
2.3. Induction of experimental diabetes
After fasting for 18 h, diabetes was induced by intraperitoneal
injection of streptozotocin (Sigma, St. Louis, MO, USA) dis-
solved in 0.1 M cold sodium citrate buffer, pH 4.5, at a dose
of 50 mg/kg.
9
The control rats received the vehicle alone.
The animals were allowed to drink 5% glucose solution over-
night to overcome the drug induced hypoglycemia. After 96 h
of streptozotocin injection, rats with moderate diabetes having
glycosuria and hyperglycemia (blood glucose range of above
250 mg/dl) were considered as diabetic rats and used for the
experiment.
2.4. Isolation of rat pancreatic islets
Pancreatic islets from adult female albino rats
10
were isolated
according to the collagenase digestion technique.
11
Batches
of five islets were picked up using stereomicroscope and trans-
ferred into small tubes each containing 1 ml KRH buffer sup-
plemented with 0.5% bovine serum albumin and glucose either
3 mmol/l (basal concentration) or 16.7 mmol/l (stimulatory
concentration) and the test agent under study was added.
The tubes were covered and incubated at 37 C in a shaking
water bath for 1 h with intermittent hand shaking every
15 min. At the end of incubation period the tubes were trans-
ferred into ice bath and mixed with vortex mixer and then ali-
quots of 0.5 ml were for insulin determination.
2.5. Experimental design in vivo
The rats were divided into four groups, each of six animals, as
follows:
Group I: Normal control rats.
Group II: STZ-induced diabetic control rats.
Group III: Diabetic rats given glimiperide (10 mg/kg) in
aqueous solution daily using an intragastric tube for 14 days.
Group IV: Diabetic rats given A. vera (10 ml/kg) daily
using an intragastric tube for 14 days.
After 14 days of the treatment, Blood samples were col-
lected from the retro – orbital venous plexus following the
technique described by Coccheto and Bjornsson (1983).
12
Briefly, rats were subjected to light ether anesthesia then blood
was collected using heparinized microhematocrit capillary
tubes into Wassermann tubes. Serum was separated by centri-
fugation at 3000 rpm for 10–15 min for the determination of
serum glucose, insulin, SOD and MDA levels. For the assess-
ment of blood GSH level blood samples were hemolyzed by
the addition of cold distilled water.
2.6. Experimental in vitro
Using isolated rat pancreatic islets, two main groups of exper-
iments were performed to study the effects of A. vera (20 lg/
ml) and glimiperide (10 lmol/l) on basal (3 mmol/l glucose)
and stimulated- insulin secretion (16.7 mmol/l glucose).
2.7. Determination of serum glucose and insulin
Fasting serum glucose was estimated by glucose oxidase meth-
od.
13
Insulin in samples, either from in vitro or in vivo experi-
ments was estimated using Enzyme Linked Immunosorbent
Assay (ELISA).
14
2.8. Determination of serum malondialdehyde (MDA),
superoxide dismutase (SOD) and glutathione (GSH)
Serum MDA was estimated by the method of Mihara and
Uchiyama (1978).
15
SOD activity was measured based on the
ability of the enzyme to inhibit the autoxidation process of
pyrogallol method of Marklund and Marklund (1974).
16
Glu-
tathione was estimated by the method of Beutler et al. (1963).
17
2.9. Statistical analysis
Data were expressed as the mean ± standard error of the mean
(s.e.m); and comparison between the different treatments was car-
ried out using one way ANOVA followed by Tukey–Kramer mul-
tiple comparisons test. Significance was accepted at p < 0.05.
3. Results
3.1. Effect of two weeks daily dose administration of A. vera and
glimiperide on serum glucose and insulin levels of streptozotocin-
induced diabetic male rats
Table 1 shows the level of serum glucose and serum insulin in
normal and STZ induced diabetic rats. There was a significant
8 A.M.H. Abo-Youssef, B.A.S. Messiha
elevation in serum glucose level and significant decrease in ser-
um insulin level during diabetes when compared with corre-
sponding control group.
Administration of A. vera and glimiperide tended to bring
the values of serum glucose and insulin levels to near normal.
3.2. Effect of two weeks daily dose administration of A. vera and
glimiperide on serum MDA, blood GSH and blood SOD levels of
streptozotocin-induced diabetic male rats
Results are graphically illustrated in Figs. 1–3. There was a sig-
nificant increase in serum MDA and significant decrease in
blood GSH and SOD during diabetes as compared to
corresponding control group. Administration of A. vera gel ex-
tract and glimiperide significantly decreased serum MDA level
and significantly elevated blood GSH and SOD levels.
3.3. Effect of A. vera and glimiperide on basal and stimulated-
insulin secretion from isolated pancreatic islets of female rats
Results are graphically illustrated in Figs. 4 and 5. These
graphs show the effect of A. vera and glimiperide on basal
and stimulated- insulin secretion from isolated pancreatic islets
of female rats.
Both A. vera and glimiperide significantly raised both basal
and stimulated- insulin secretion as compared to the normal
control value (3 mmol/l glucose).
Table 1 Effect of two weeks daily dose administration of Aloe vera and glimiperide on serum glucose and insulin levels of
streptozotocin-induced diabetic male rats.
Groups Serum glucose level (mg/dl) Serum insulin level (ng/ml)
Normal control 82.24 ± 5.17 7.05 ± 0.76
Diabetic control 331.88 ± 29.72
*
2.82 ± 0.66
*
Diabetic + A. vera 93.66 ± 26.92
a
6.78 ± 0.98
a
Diabetic + glimiperide 117.43 ± 21.96
a
6.26 ± 0.65
a
Values are given as mean ± s.e.m for groups of six animals each.
Values are statistically significant at p < 0.05.
Diabetic control was compared to normal control rats.
Diabetic + A. vera and diabetic + glimiperide were compared with diabetic control.
*
Significantly different from the normal control.
a
Significantly different from the diabetic control.
Values are given as mean ± s.e.m for groups of six animals each.
Values are statistically significant at p < 0.05.
Diabetic control was compared to normal control rats.
Diabetic + Aloe vera and diabetic + glimiperide were compared with diabetic control.
* significantly different from the normal control.
a significantly different from the diabetic control.
Figure. 1 Effect of two weeks daily dose administration of Aloe
vera and glimiperide on serum MDA level of streptozotocin-
induced diabetic male rats.
Figure. 2 Effect of two weeks daily dose administration of Aloe
vera and glimiperide on blood GSH level of streptozotocin-
induced diabetic male rats.
Beneficial effects of Aloe vera in treatment of diabetes: Comparative in vivo and in vitro studies 9
4. Discussion
Results of the present investigation strongly suggest an antidi-
abetic potential for aloe. In-vivo results show decreased serum
glucose and increased serum insulin levels in aloe-treated rats
compared to diabetic controls (Table 1). In agreement, previ-
ous authors claimed for such antidiabetic effect for aloe extract
on experimental animals.
18,19
Our in vitro results support this
finding, where aloe was found to increase the rate of insulin
secretion from pancreatic islets (Figs. 4 and 5).
Many explanations were suggested for this antidiabetic ef-
fect of aloe. The first explanation is the potent antioxidant ef-
fect of aloe extract. Aloe is long known to have antioxidant
potential via suppression of free radical formation and
enhancement of cellular thiol status.
20–22
It is also reported
to stimulate glutathione-S-transferase enzyme activity.
23
Our
results strongly supported the antioxidant potential of aloe,
where it was found to suppress elevated serum MDA levels
and increase blood GSH and SOD levels. Recent approaches
focus on the role of oxidative stress in pancreatic beta cell
damage.
8,24
That is, oxidative stress is involved as a causative
factor in the pathogenesis of diabetes, and hence antioxidants
like aloe may have a true antidiabetic effect via antioxidant
potential.
The anti-inflammatory potential of aloe may be the second
explanation for its antidiabetic effect. Diabetes may be consid-
ered as an inflammatory disease where inflammation partici-
pates in the progression of diabetes, where tumor necrosis
factor-a was found to decrease peripheral insulin sensitivity.
25
Many authors claimed for the anti-inflammatory potential of
aloe due to many of its components like emodin and man-
nose-6-phosphate.
26,27
It was reported that the anti-inflamma-
tory effect of aloe extract is comparable to that of
hydrocortisone.
28
Finally, aloe may act as a hypoglycemic agent through po-
tent inhibition of pancreatic a-amylase activity.
29
This action
decreases starch breakdown and offers good postprandial gly-
cemic control.
These findings are promising for further clinical studies on
aloe extract or extract components in the management of dia-
betes mellitus.
Figure. 4 Effect of Aloe vera and glimiperide on basal insulin
secretion from isolated pancreatic islets of female rats.
Figure. 3 Effect of two weeks daily dose administration of Aloe
vera and glimiperide on blood SOD level of streptozotocin-
induced diabetic male rats.
Figure. 5 Effect of Aloe vera and glimiperide on stimulated-
insulin secretion from isolated pancreatic islets of female rats.
10 A.M.H. Abo-Youssef, B.A.S. Messiha
5. Conflict of interest
None.
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... A. vera has also been shown to have a potential effect on glycaemic control, reducing hyperglycaemia and hypercholesterolaemia in people with DM. Aloeresin-A, an active compound of A. vera, inhibits α-glucosidase activity and reduces α-glucosidase and intestinal glucose absorption [36][37][38][39][40][41]. ...
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Full-text available
  • Jan 2025
This study presents an investigation of the chemical composition of Aloe vera leaf tissue with a focus on the spatial distribution of compounds. The composition was studied using two mass spectrometry imaging techniques: silver-109 nanoparticles assisted laser desorption/ionization mass spectrometry imaging (¹⁰⁹AgNPs-LDI-MSI) and laser ablation-remote atmospheric pressure photoionization/chemical ionization mass spectrometry imaging (LARAPPI/CI-MSI) and the identification was aided by ultra-high-performance liquid chromatography and ultra-high-resolution mass spectrometry (UHPLC-UHRMS) analysis. The results showed an abundance of phenolic compounds with antioxidant, antimicrobial, and anti-inflammatory properties, making it a beneficial food additive and food packaging material. Analysis of the results of mass spectrometry imaging provided information about the potential changes in metabolic pathway expression in different regions of the leaf.
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The Role of Herbs in the Treatment of Diabetes
Chapter
  • Jul 2024
The Chemistry inside Spices & Herbs: Research and Development brings comprehensive information about the chemistry of spices and herbs with a focus on recent research in this field. Experts in phytochemistry have contributed reviews with the aim to give the reader deep knowledge about phytochemical constituents in herbal plants and their benefits. The contents include reviews on the biochemistry and biotechnology of spices and herbs, herbal medicines, biologically active compounds and their role in therapeutics among other topics. Chapters which highlight natural drugs and their role in different diseases and special plants of clinical significance are also included. Volume 3 covers several topics: the treatment of Polycystic Ovary Syndrome (PCOS), managing rheumatoid arthritis and related inflammatory conditions, orchid-derived natural flavoring and therapeutic agent Vanillin, Silymarin's utility in treating hepatic diseases, phytochemistry and pharmacological activities of Hygrophila spinosa, pharmacological and chemical aspects of Tulsi, Combretum caffrum as a potential anticancer molecule, and the roles of herbs in treating diabetes. This book is an ideal resource for scholars (in life sciences, phytomedicine and natural product chemistry) and general readers who want to understand the importance of herbs, spices and traditional medicine in pharmaceutical R&D and clinical research.
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Ethnobotanical assessment of antidiabetic medicinal plants in District Karak, Pakistan
Article
Full-text available
  • Apr 2024
Background Diabetes is a leading health disorder and is responsible for high mortality rates across the globe. Multiple treatment protocols are being applied to overcome this morbidity and mortality including plant-based traditional medicines. This study was designed to investigate the ethnomedicinal status of plant species used to treat diabetes in District Karak, Pakistan. Materials and methods A semi-structured survey was created to collect data about traditionally used medicinal plants for diabetes and other ailments. The convenience sampling method was applied for the selection of informants. The collected data was evaluated through quantitative tools like frequency of citation (FC), relative frequency of citation (RFC), informant consensus factor (FIC), fidelity level (FL), and use value (UV). Results A total of 346 local informants were selected for this research. Out of them, 135 participants were men and 211 participants were women. Overall 38 plant species belonging to 29 plant families were used to treat diabetes. The most dominant plant family was Oleaceae having 11 species. Powder form (19%) was the most recommended mode of preparation for plant-based ethnomedicines. Leaves (68%) were the most frequently used parts followed by fruit (47%). The highest RFC was recorded for Apteranthes tuberculata (0.147). The maximum FL was reported for Apteranthes tuberculata (94.4) and Zygophyllum indicum (94.11) for diabetes, skin, and wounds. Similarly, the highest UV of (1) each was found for Brassica rapa, Melia azedarach, and Calotropis procera. Based on documented data, the reported ailments were grouped into 7 categories. The ICF values range between 0.89 (diabetes) to 0.33 (Cardiovascular disorders). Conclusion The study includes a variety of antidiabetic medicinal plants, which are used by the locals in various herbal preparations. The species Apteranthes tuberculata has been reported to be the most frequently used medicinal plant against diabetes. Therefore, it is recommended that such plants be further investigated in-vitro and in-vivo to determine their anti-diabetic effects.
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Review on Diabetic Complications and their Management by Flavonoids and Triterpenoids
Article
  • Dec 2023
Diabetes mellitus, together with its numerous consequences, is rapidly becoming a major health issue. Natural products are secondary metabolites found in plants that have a wide range of biological functions. The development of anti-diabetic medications derived from natural compounds, particularly those derived from plants having a documented folk-use history in the treatment of diabetes, is gaining traction. Many studies have shown the usefulness of natural flavonoids with hypoglycemic properties in the management of diabetic problems, along with their advantages. This paper describes the mechanisms of action of several natural flavonoids whose hypoglycemic effects have been confirmed. Comprehensive lifestyle treatments can help those at high risk of diabetes to avoid or delay the start of the disease, according to the results of randomized controlled trials. Terpenoids are a type of natural substance that have been identified as an anti-diabetic agent in various studies. Some of them are in various phases of preclinical and clinical testing to conclude whether they can be used as anti-diabetic drugs. These compounds can block the enzymes involved in insulin resistance, facilitate glucose metabolism, and positively affect plasma glucose and insulin levels. By blocking multiple pathways implicated in diabetes and its consequences, flavonoids and triterpenes can operate as potential agents in the treatment of diabetic retinopathy, neuropathy, and nephropathy, as well as poor wound healing. However, there have been few attempts to investigate the biological effects of triterpenes and clinical research investigating their use in the treatment of diabetes. As a result, it is critical to pay close attention to these chemicals' therapeutic potential and to contribute fresh information to the scientific community. This review focuses on current advancements in flavonoids and triterpenes chemistry, derivatives, biological interventions, and therapeutic applications, with a focus on diabetes and related illnesses.
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Protective Effect of Aloe Extract against the Cytotoxicity of 1,4Naphthoquinone in Isolated Rat Hepatocytes Involves Modulations in Cellular Thiol Levels
Article
Full-text available
  • May 2002
  • Pharmacol Toxicol
: Aloe is a familiar ingredient in a wide range of health care and cosmetic products and has been reported to possess various physiological effects, antioxidative, anticarcinogenic, antiinflammatory and laxative. Aloe has also been reported to have an effect on liver function. The cytoprotective effect of aloe extract against 1,4-naphthoquinone-induced hepatotoxicity was evaluated in primary cultured rat hepatocytes. After exposure to 1,4-naphthoquinone (100 μM), a decrease in cell viability measured as >60% lactate dehydrogenase depletion was induced. Cellular glutathione (GSH) and protein-SH levels were also significantly decreased in a time-dependent manner. However addition of aloe extract resulted in a dose-dependent improvement of these effects. This cytoprotective effect of aloe could be attributed to its inhibition of GSH and protein-SH depletions. The effect of the aloe extracts were also dose-dependent. Addition of diethyl maleate (1 mM), a cellular glutathione-depleting agent, to hepatocytes treated with both 1,4-naphthoquinone and aloe extract, induced depletion of GSH, but did not affect protein-SH or lactate dehydrogenase. These results suggest that the 1,4-naphthoquinone-induced toxicity in rat hepatocytes was inhibited by aloe extract, and that this protective effect was due to the maintenance of cellular thiols, especially protein-SH.
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Protective effect of Aloe extract against the cytotoxicity of 1,4-Naphthoquinone in isolated rat hepatocytes involves modulation of cellular thiol levels
Article
Full-text available
  • Jan 2002
Aloe is a familiar ingredient in a wide range of health care and cosmetic products and has been reported to possess various physiological effects, antioxidative, anticarcinogenic, antiinflammatory and laxative. Aloe has also been reported to have an effect on liver function. The cytoprotective effect of aloe extract against 1,4-naphthoquinone-induced hepatotoxicity was evaluated in primary cultured rat hepatocytes. After exposure to 1,4-naphthoquinone (100 mM), a decrease in cell viability measured as ±60% lactate dehydrogenase depletion was induced. Cellular glutathione (GSH) and protein-SH levels were also significantly decreased in a time-dependent manner. However addition of aloe extract resulted in a dose-dependent improvement of these effects. This cytoprotective effect of aloe could be attributed to its inhibition of GSH and protein-SH depletions. The effect of the aloe extracts were also dose-dependent. Addition of diethyl maleate (1 mM), a cellular glutathione-depleting agent, to hepatocytes treated with both 1,4-naphthoquinone and aloe extract, induced depletion of GSH, but did not affect protein-SH or lactate dehydrogenase. These results suggest that the 1,4-naphthoquinone-induced toxicity in rat hepatocytes was inhibited by aloe extract, and that this protective effect was due to the maintenance of cellular thiols, especially protein-SH.
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Potent α-amylase inhibitory activity of Indian Ayurvedic medicinal plants
Article
Full-text available
  • Jan 2011
  • BMC Compl Alternative Med
Background Indian medicinal plants used in the Ayurvedic traditional system to treat diabetes are a valuable source of novel anti-diabetic agents. Pancreatic α-amylase inhibitors offer an effective strategy to lower the levels of post-prandial hyperglycemia via control of starch breakdown. In this study, seventeen Indian medicinal plants with known hypoglycemic properties were subjected to sequential solvent extraction and tested for α-amylase inhibition, in order to assess and evaluate their inhibitory potential on PPA (porcine pancreatic α-amylase). Preliminary phytochemical analysis of the lead extracts was performed in order to determine the probable constituents. Methods Analysis of the 126 extracts, obtained from 17 plants (Aloe vera (L.) Burm.f., Adansonia digitata L., Allium sativum L., Casia fistula L., Catharanthus roseus (L.) G. Don., Cinnamomum verum Persl., Coccinia grandis (L.) Voigt., Linum usitatisumum L., Mangifera indica L., Morus alba L., Nerium oleander L., Ocimum tenuiflorum L., Piper nigrum L., Terminalia chebula Retz., Tinospora cordifolia (Willd.) Miers., Trigonella foenum-graceum L., Zingiber officinale Rosc.) for PPA inhibition was initially performed qualitatively by starch-iodine colour assay. The lead extracts were further quantified with respect to PPA inhibition using the chromogenic DNSA (3, 5-dinitrosalicylic acid) method. Phytochemical constituents of the extracts exhibiting≥ 50% inhibition were analysed qualitatively as well as by GC-MS (Gas chromatography-Mass spectrometry). Results Of the 126 extracts obtained from 17 plants, 17 extracts exhibited PPA inhibitory potential to varying degrees (10%-60.5%) while 4 extracts showed low inhibition (< 10%). However, strong porcine pancreatic amylase inhibitory activity (> 50%) was obtained with 3 isopropanol extracts. All these 3 extracts exhibited concentration dependent inhibition with IC50 values, viz., seeds of Linum usitatisumum (540 μgml-1), leaves of Morus alba (1440 μgml-1) and Ocimum tenuiflorum (8.9 μgml-1). Acarbose as the standard inhibitor exhibited an IC50 (half maximal inhibitory concentration)value of 10.2 μgml-1. Phytochemical analysis revealed the presence of alkaloids, tannins, cardiac glycosides, flavonoids, saponins and steroids with the major phytoconstituents being identified by GC-MS. Conclusions This study endorses the use of these plants for further studies to determine their potential for type 2 diabetes management. Results suggests that extracts of Linum usitatisumum, Morus alba and Ocimum tenuiflorum act effectively as PPA inhibitors leading to a reduction in starch hydrolysis and hence eventually to lowered glucose levels.
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Clinical and experimental evidence for oxidative stress as an exacerbating factor of diabetes mellitus
Article
Full-text available
  • Jan 2011
  • J CLIN BIOCHEM NUTR
The involvement of reactive oxygen species in various diseases has been demonstrated almost in vitro or in animal studies and clinical studies supporting the involvement of reactive oxygen species are very few. Bilirubin has been recognized as an important antioxidant and also shown to have an inhibitory effect on the activity of NADPH oxidase, which may be an important source for superoxide production in various tissues. When the prevalence of vascular complcations was compared in diabetic patients with and without a congenital hyperbilirubinemia (Gilbert syndrome), the prevalence of retinopathy, macroalbuminuria and coronary artery disease in patients with Gilbert syndrome was about 20% of that in those without Gilbert syndrome. For study of lifestyle-related diseases, the Fukuoka Cohort was constructed from 2003 to 2009 in Kyushu area in Japan, which contains a total of 12,949 persons. Cross-sectional study of the Fukuoka Cohort revealed an inverse relation between serum bilirubin level and the prevalence of type 2 diabetes mellitus. A precursor of bilirubin, biliverdin-treated db/db mice exhibited less albuminuria and nephropathic changes. These effects were paralleled with normalization of oxidative stress markers and expression of NAD(P)H oxidase subunits in kidney. These results suggested that oxidative stress is an exacerbating factor of type 2 diabetes mellitus and that antioxidant therapies are of value to diabetic nephropathy.
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Involvement of the Superoxide Anion Radical in the Autoxidation of Pyrogallol and a Convenient Assay for Superoxide Dismutase
Article
  • Mar 2005
  • Eur J Biochem
The autoxidation of pyrogallol was investigated in the presence of EDTA in the pH range 7.9–10.6. The rate of autoxidation increases with increasing pH. At pH 7.9 the reaction is inhibited to 99% by superoxide dismutase, indicating an almost total dependence on the participation of the superoxide anion radical, O2·−, in the reaction. Up to pH 9.1 the reaction is still inhibited to over 90% by superoxide dismutase, but at higher alkalinity, O2·− -independent mechanisms rapidly become dominant. Catalase has no effect on the autoxidation but decreases the oxygen consumption by half, showing that H2O2 is the stable product of oxygen and that H2O2 is not involved in the autoxidation mechanism. A simple and rapid method for the assay of superoxide dismutase is described, based on the ability of the enzyme to inhibit the autoxidation of pyrogallol. A plausible explanation is given for the non-competitive part of the inhibition of catechol O-methyltransferase brought about by pyrogallol.
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Electrophysiological effects of osmotic cell shrinkage in rat pancreatic β-Cells
Article
  • May 2010
Electrical and secretory activity in the pancreatic β-cell can be elicited by hypotonic cell swelling, due largely to activation of a volume-regulated anion channel (VRAC) leading to depolarisation and electrical activity. However, β-cell responses to cell shrinkage are less well characterised. The present study has examined the effects of osmotic cell shrinkage on rat pancreatic β-cells. Electrical activity and whole-cell current were studied in isolated β-cells using the perforated patch and conventional whole-cell recording techniques. Insulin release was measured using intact islets by radioimmunoassay. Exposure to a 33% hypertonic bath solution resulted in an initial depolarisation and a period of electrical activity. In several cases, this depolarisation was transient and was followed by a hyperpolarisation. A similar pattern was observed with insulin release. In voltage-clamp experiments, osmotic shrinkage resulted in activation of a non-selective cation channel (NSCC) sensitive to inhibition by flufenamic acid and Gd3+. It is suggested that activation of this NSCC is responsible for the depolarisation evoked by hypertonic media. The secondary hyperpolarisation is likely to be the result of inhibition of VRAC activity. These opposing ionic effects could underlie the biphasic effect on insulin release following exposure to hypertonic media.
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Oxidative Stress and Diabetic Complications
Article
  • Oct 2010
  • CIRC RES
Oxidative stress plays a pivotal role in the development of diabetes complications, both microvascular and cardiovascular. The metabolic abnormalities of diabetes cause mitochondrial superoxide overproduction in endothelial cells of both large and small vessels, as well as in the myocardium. This increased superoxide production causes the activation of 5 major pathways involved in the pathogenesis of complications: polyol pathway flux, increased formation of AGEs (advanced glycation end products), increased expression of the receptor for AGEs and its activating ligands, activation of protein kinase C isoforms, and overactivity of the hexosamine pathway. It also directly inactivates 2 critical antiatherosclerotic enzymes, endothelial nitric oxide synthase and prostacyclin synthase. Through these pathways, increased intracellular reactive oxygen species (ROS) cause defective angiogenesis in response to ischemia, activate a number of proinflammatory pathways, and cause long-lasting epigenetic changes that drive persistent expression of proinflammatory genes after glycemia is normalized ("hyperglycemic memory"). Atherosclerosis and cardiomyopathy in type 2 diabetes are caused in part by pathway-selective insulin resistance, which increases mitochondrial ROS production from free fatty acids and by inactivation of antiatherosclerosis enzymes by ROS. Overexpression of superoxide dismutase in transgenic diabetic mice prevents diabetic retinopathy, nephropathy, and cardiomyopathy. The aim of this review is to highlight advances in understanding the role of metabolite-generated ROS in the development of diabetic complications.
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Aloe vera gel alleviates cardiotoxicity in streptozocin-induced diabetes in rats
Article
  • Jan 2010
  • J PHARM PHARMACOL
Persistent hyperglycaemia results in oxidative stress along with the generation of oxygen free radicals and appears to be an important factor in the production of secondary complications in diabetes. The aim of this work was to evaluate markers of oxidative stress in heart tissue along with the protective, antioxidant and antidiabetic activity of 30%Aloe vera gel in diabetic rats. Streptozocin was given as a single intravenous injection and 30%Aloe vera gel was given in two doses for 20 days, orally. Blood glucose, glycosylated haemoglobin, blood reduced glutathione, serum lactate dehydrogenase and serum creatine kinase levels were measured on day 21 after drug treatment. Heart rate and mean blood pressure were recorded at the end of the study. Different biochemical variables were evaluated in the heart tissue, including thiobarbituric acid reactive substance (TBARS), reduced glutathione, superoxide dismutase and catalase in diabetic and in Aloe vera-treated diabetic rats. In streptozocin diabetic rats, the TBARS level was increased significantly, superoxide dismutase and reduced glutathione significantly decreased, and the catalase level was significantly increased. Aloe vera 30% gel (200 mg/kg) treatment in diabetic rats reduced the increased TBARS and maintained the superoxide dismutase and catalase activity up to the normal level. Aloe vera gel increased reduced glutathione by four times in diabetic rats. Aloe vera gel at 200 mg/kg had significant antidiabetic and cardioprotective activity.
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