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64
Cell. Mol. Biol. (ISSN: 1165-158X)
1. Introduction
Sepsis is a heterogeneous syndrome characterized by
a highly intricate interplay between various biological
networks. Our understanding related to the key mecha-
nisms involved in the pathogenesis of sepsis has tremen-
chemicals is interconnected with pathophysiological me-
pleiotropic multifaceted mediators that connect and rewire
[1, 2]. It
-
fore, in sepsis, crosstalks between complement and coagu-
lation pathways contribute to the disease aggressiveness
[3, 4].
Sepsis is an important cause of acute lung injury and its
more severe form, acute respiratory distress syndrome [5].
Acute lung injury caused by sepsis a major reason of re-
-
bility following damage to alveolar epithelial and endothe-
lial cells due to sepsis can lead to pulmonary edema [6].
against tissue damage caused by sepsis [7].
-
cule, is an endogenous hormone that is secreted at night
Original Article
Investigation of the eects of melatonin on lung tissue through the NLRP3/TLR2/
NEK7 pathway in an experimental endotoxemia model
Arif Osman Tokat1*, Osman Öztürk2, Aslı Okan3, Sümeyye Uçar4, Ece Eroğlu5, Züleyha Doğanyıği3,
Mert Ocak6, Şükrü Ateş7, Seher Yilmaz7
1 Yozgat Bozok University, Faculty of Medicine, Department of Thoracic Surgery, Yozgat, Turkey
2 Yozgat Bozok University, Faculty of Medicine, Department of Child Health and Diseases, Yozgat, Turkey
3Yozgat Bozok University, Faculty of Medicine, Department of Histology and Embryology, Yozgat, Turkey
4 Erciyes University, Faculty of Medicine, Department of Anatomy, Kayseri, Turkey
5 Yozgat Bozok University, Faculty of Medicine,Yozgat,Turkey
6 Ankara University, Faculty of Dentistry, Department of Anatomy, Ankara, Turkey
7 Yozgat Bozok University, Faculty of Medicine, Department of Anatomy, Yozgat, Turkey
Journal Homepage: www.cellmolbiol.org
Cellular and Molecular Biology
E-mail address: d.kahrizi@cellmolbiol.org (D. Kahrizi).
Doi: 0.14715/cmb/2024.70.10.10
Article Info Abstract
Article history:
Received: February 08, 2024
Accepted: June 09, 2024
Published: October 31, 2024
-
-
-
Keywords:
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65
E ec t s o f me l at o ni n on L PS - in d uc e d l u ng i nj u ry. Cell. Mol. Biol. 2024, 70(10): 64-71
from the gland called pineal or pineal and plays contri-
butory role in the modulation of various biological func-
tions such as sleep, reproduction, immunity and biological
blood pressure regulation [8, 9]-
crine rhythm, regulating immune functions, and protection
[10, 11].
Melatonin has pleiotropic roles in the regulation of dif-
ferent cellular process including the sleep-wake cycle and
its receptors are widely distributed. A study conducted in
2014 reported that melatonin regulates mitogen-activa-
mediators in microbial-induced sepsis [12]. Melatonin ap-
pears to be associated with the sepsis state through its an-
[13, 14]
Melatonin intervention may contribute to better survival
in a septic animal model [15–17]. In this regard, melatonin
septic shock due to sepsis.
-
sembled in the cytosol in response to damage-associated
and pathogen-associated stimuli as well as other danger
[18].
-
tion of several cellular processes that stimulate additional
-
tions such as sepsis [19]. In addition, it is known that the
-
lated lung injury [20]
-
[21]
-
ted during Staphylococcus aureus-associated septic infec-
[20].
-
pattern recognition receptors covering various antigenic
determinants [22]
following sepsis-induced acute lung injury [23]
-
gnizes a variety of microorganisms, such as viruses, fungi
[24] -
conditions [25]-
from failure of multiple organs [26].
samples, including live specimens and various solid or
such as imaging and analysis of soft tissues and bones.
animals, contributing to its widespread use in the health
diagnostic method in animal studies, particularly in the re-
search and development of new drugs and agents [27, 28].
-
histopathological and immunohistochemical evaluation,
2. Material and Methods
2.1. Experimental groups
-
the formal approval (Ethical committee decision number:
-
-
mental animals had free access to standard food and tap
water. All procedures were strictly in compliance with the
-
-
nistering ketamine hydrochloride (50 mg/kg intramuscular
mg/kg i.m./i.p.).
In our research design, Sprague-Dawley adult male rats
= 7), and these groups are listed below. Administration
-
ding to the previously reported literature [1, 29, 30]. Addi-
possible morphological or symptomatic change. In the stu-
dy, all i.p. injection and gavage applications were perfor-
med at similar timings of the day. Since the current study
tissue in sepsis, melatonin was applied to the 3rd and 4th
-
tion of melatonin application in the 4th group, at the end
intraperitoneally after 1 hour of application.
Group 1 (Control group, n=7): 1 ml physiological
-
nistered to the animals via i.p. every day for 10 days.
Group 2 (Melatonin group, n=7): Animals were
given 1 ml i.p. melatonin at a dose of 10 mg/kg. It was
applied daily for 10 days [30].
Group 3 (LPS group, n=7):
a dose of 1 ml at a dose of 30 mg/kg was administered
to the animals i.p. as a single dose on the last day of the
[1].
Group 4 (Melatonin + LPS group, n=7): Animals
were given 10 mg/kg melatonin 1 ml i.p. was administered
of 1 ml.
2.2. Dissection of lung tissue of experimental groups
anesthesia, and lung tissue was dissected.
66
E ec t s o f me l at o ni n on L PS - in d uc e d l u ng i nj u ry. Cell. Mol. Biol. 2024, 70(10): 64-71
2.5. ELISA method
For biochemical analyses, tissue samples were taken
manufacturer was performed to determine rat MDA levels.
2.6. Micro CT method
-
-
red for each sample using a Bruker Skyscan 1275 appa-
ratus located in Kontich, Belgium. For imaging through
ms, rotation step of 0.2, and a 360-degree rotation within
sagittal images for each sample were scrutinized utilizing
Dataviewer software from Skyscan, Kontich, Belgium.
For three-dimensional (3D) volumetric visualization and
-
laar, Belgium, was utilized.
2.7. Statistical analysis
-
dings were compared by two-way analysis of variance
intensity (%) were compared by one-way analysis of va-
p<0.05 was
used for multiple comparisons between groups.
3. Results
3.1. Histological analysis results
Bronchioles, alveolar, and pulmonary arteries were ob-
served in normal histological structure in the lung tissues
of the control and melatonin groups. Hemorrhage, cellular
-
1, Figure 1A).
3.2. Immunohistochemical analysis
2.3. Histological analysis
-
-
tion, the tissues were left under running water overnight.
series (70%, 80%, and 96%) for one day each. After being
left in 100% alcohol for 3 hours, the process was made
-
-
-
according to whether 0 was none, 1 was mild, 2 was mode-
rate, and 3 was severe [31].
2.4. Immunohistochemical analysis
-
cience, E-AB) were detected in the lung tissues taken from
-
dase method. -70161) immunoreactivities were detected
by immunohistochemical analysis [29, 32]. After deparaf-
-
-
-
o
it was incubated with biotinylated goat anti-polyvalent
o -
o-
-
-
terstaining of the sections was done with Gill III Hema-
of the sections was done by passing through a series of
successively increasing alcohol dilutions and sealing with
an Olympus BX53 light microscope. Immunoreactivity
Bethesda, Maryland).
Groups Bleeding Cellular Inltration Alveolar wall thickening
0.22±0.09 0.22±0.09 0.11±0.09
0.22±0.19 0.33±0.16 0.5±0.16
1.22±0.38ab 1.05±0.09ab 1.83±0.28ab
0.66±0.28c0.72±0.09a0.66±0.28ac
apbpcp<0.05
Table 1. Damage rates observed in lung tissues.
67
E ec t s o f me l at o ni n on L PS - in d uc e d l u ng i nj u ry. Cell. Mol. Biol. 2024, 70(10): 64-71
-
-
group (Figures 1A and 1B).
3.3. ELISA method results
However, these levels were found to be reduced in the tis-
3.4. Micro CT method results
compared to the other groups. Object volume and Object
(Figure 3B).
4. Discussion
In this study, the immunoreactivity of melatonin to
-
compared to the control group; It was determined that
-
-
-
from immune cells, especially monocytes and macro-
phages [33]
Fig. 1.
-
ties (B).
mean±SD. ap
control group, bp
cp
Fig. 2. MDA levels of lung tissues. Data shown in bar graphs are
p-
ference (ns: p>0.05; **p<0.01; ***p<0.001).
Fig. 3.
-
pressed as mean±SD. p-
rence (ns: p>0.05; *p<0.05; **p<0.01; ***p<0.001).
68
E ec t s o f me l at o ni n on L PS - in d uc e d l u ng i nj u ry. Cell. Mol. Biol. 2024, 70(10): 64-71
to the cell generating the host response [34]. Following sti-
-
macrophages, and lymphocytes and enter the systematic
circulation [35]
are observed [36]
as melatonin may contribute to the reduction of organ da-
mage in sepsis [37].
In similar studies in the literature, it has been shown by
[38, 39]-
tion triggers intracellular transduction that results in acti-
[40].
active immune response and prevent damaging immu-
study in which sepsis treatment was performed with an
-
-
-
sis model in mice [41]
sepsis patients can be suggested as a powerful therapeutic
-
-
Arg753Gln polymorphisms and sepsis [42]. Accordingly,
-
tion may be a promising approach in the treatment of acute
[43]. In the literature, it has been shown that animals with
induced organ damage and shock [19, 44]. Suppression
platelet activation [45]
can accelerate caspase-1 maturation, leading to acute lung
[46]. In this regard, therapeutic approaches that limit the
-
activate this molecule by binding to the leucine-rich repeat
region [47, 48]. In addition, in sepsis-related acute lung
-
tion [20]
group as compared to the control group. A study in 2023
-
[49].
-
decrease in the lungs of septic mice given melatonin
agents. Based on these data, therapeutic approaches such
as melatonin may reduce the severity of sepsis-induced
lung injury due to sepsis model [50, 51]. Melatonin can
-
dings are consistent with the literature, bleeding, cellular
-
red to the control and melatonin groups. Administration of
-
dant substance, may have protective properties against
[7] -
ties of melatonin in sepsis have been attributed to reasons
shown that with the application of a melatonin protective
the lung [13]
activated [55]. Our study reveals that melatonin adminis-
-
ging in small animal disease models is very important in
elucidating the etiology of various diseases and in develo-
[56]. Bell et
al., measured mouse lung tissue volume in an interstitial
-
ported that the results were compatible with histopatho-
invasive method [57]
study, it was reported that the total air volume of the lung
-
luation of the lung ultrastructure and its changes during
[58]. In another similar
-
caused a decrease in total lung volüme [59]. In our study,
-
that were correlated with the results obtained in histopa-
thological and immunohistochemical analyses. In addi-
tion, the results we obtained and the information in the
literature support each other.
5. Conclusion
-
-
treatment groups approached control group. However,
performing only MDA results and immunohistochemical
analyses using the Eliza method can be considered a limi-
tation of our research. Future research works should be
69
E ec t s o f me l at o ni n on L PS - in d uc e d l u ng i nj u ry. Cell. Mol. Biol. 2024, 70(10): 64-71
focused on a detailed mapping of protein networks related
Conict of interests
preparation.
Consent for publications
publication.
Ethics approval and consent to participate
(Ethical committee decision number: 22/277).
Informed consent
Availability of data and material
Authors' contributions
answering reviewers.
Funding
Acknowledgements
-
-
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