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MESNA (2-Mercaptoethanesulfonate) Attenuates Brain, Heart, and Lung Injury

Induced by Carotid Ischemia‑Reperfusion in Rats

Article in Nigerian Journal of Clinical Practice · August 2023

DOI: 10.4103/njcp.njcp_654_22

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 Original Article

MESNA (2‑Mercaptoethanesulfonate) Attenuates Brain, Heart, and

Lung Injury Induced by Carotid Ischemia‑Reperfusion in Rats
M Mercan, AÖ Şehirli1, Ç Gültekin2, U Chukwunyere3, S Sayıner4, S Gençosman4, Ş Çetinel5, N Abacıoğlu3

Department of Background: Ischemia‑reperfusion (I/R) causes organ dysfunction as a

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Abstract
Pharmacology, Faculty of
Pharmacy, 1Department
result of the increased formation of various reactive oxygen metabolites,
of Pharmacology, Faculty infiltration of inflammatory cells, interstitial edema, cellular dysfunction, and
of Dentistry, 2Department tissue death. Aim: The study aimed to investigate the cytoprotective effect of
nYQp/IlQrHD3i3D0OdRyi7TvSFl4Cf3VC4/OAVpDDa8KKGKV0Ymy+78= on 08/06/2023

of Surgery, Faculty of 2‑mercaptoethanesulfonate (MESNA) against tissue damage in rats exposed

Veterinary, 3Department of to carotid ischemia‑reperfusion. Materials and Methods: Twenty‑four male
Pharmacology, Faculty of Wistar albino rats were divided into four groups (n = 6): sham, carotid I/R,
Pharmacy, 4Department of
Biochemistry, Faculty of
I/R + MESNA (75 mg/kg), and I/R + MESNA (150 mg/kg) groups. To induce
Veterinary Medicine, Near ischemia in rats, the carotid arteries were ligated with silk sutures for 10 min;
East University, Near East the silk suture was then opened, and 1 h reperfusion was done. MESNA (75 and
Boulevard, 99138 Nicosia, 150 mg/kg) was administered intraperitoneally 30 min before ischemia‑reperfusion.
North Cyprus, 5Department of Tissue samples from the animals were taken for histological examination, while
Histology and Embryology, the serum levels of some biochemical parameters were utilized to evaluate
Faculty of Medicine,
Marmara University, İstanbul,
the systemic alterations. ANOVA and Tukey’s post hoc tests were applied with
Türkiye a significance level of 5%. Results: The ischemia‑reperfusion‑induced tissue
damage as evidenced by increase in serum levels of alanine transaminase, aspartate
aminotransferase, alkaline phosphatase, malondialdehyde, lactate dehydrogenase,
and matrix metalloproteinases (MMP‑1, ‑2, ‑8) was significantly (P < 0.05–0.0001)
Received:
reversed after treatment with MESNA in a dose‑dependent manner. Treatment
27-Sep-2022; with MESNA (75 and 150 mg/kg), significantly (P < 0.05–0.0001) decreased
Revision: the I/R‑induced increase in serum tumor necrosis factor‑alpha (TNF‑α) and
05-Mar-2023; Interleukin‑1‑beta (IL‑1 β). Conclusion: The results of this study suggest that
Accepted: MESNA has a protective effect on tissues by suppressing cellular responses to
08-Mar-2023; oxidants and inflammatory mediators associated with carotid ischemia‑reperfusion.
Published:
31-Jul-2023 Keywords: Carotid artery, Ischemia, MESNA, Oxidative stress, Reperfusion

Introduction result in cerebral edema and severe neurological

dysfunctions.[2]
C arotid ischemia is a serious health concern
because of its high morbidity and mortality
rates.[1] Ischemia‑induced tissue damages are well
Reduced blood flow to the brain if left untreated
can lead to conditions such as stroke, cardiac arrest,
defined, leading first to damage of brain tissue and pulmonary embolism, and even death.[1] Reperfusion
then to pathological damage of distant organs such as
the lungs and the heart. While reperfusion is essential
Address for correspondence: Dr. M Mercan,
to reverse the effects of ischemia, it also causes Department of Pharmacology, Faculty of Pharmacy,
additional damage to tissues in severe cases. Oxidative Near East University, Near East Boulevard, 99138 Nicosia,
stress, infiltration of inflammatory cells, and disruption North Cyprus.
E‑mail: [email protected]
of the blood‑brain barrier are some of the distinct
mechanistic pathways of reperfusion injury that can This is an open access journal, and articles are distributed under the terms of the
Creative Commons Attribution‑NonCommercial‑ShareAlike 4.0 License, which allows
others to remix, tweak, and build upon the work non‑commercially, as long as
Access this article online appropriate credit is given and the new creations are licensed under the identical
Quick Response Code: terms.
Website: www.njcponline.com
For reprints contact: [email protected]

DOI: 10.4103/njcp.njcp_654_22 How to cite this article: Mercan M, Şehirli AÖ, Gültekin Ç, Chukwunyere U,
Sayıner S, Gençosman S, et al. MESNA (2‑mercaptoethanesulfonate) attenuates
brain, heart, and lung injury induced by carotid ischemia‑reperfusion in
rats. Niger J Clin Pract 2023;26:941-8.

© 2023 Nigerian Journal of Clinical Practice | Published by Wolters Kluwer ‑ Medknow 941
 Mercan, et al.: Effects of MESNA on carotid ischemia‑reperfusion in rats

increases the levels and activation of inflammatory I/R group was exposed to ischemia‑reperfusion and
cytokines such as tumor necrosis factor‑alpha (TNF‑α) treated with normal saline (intraperitoneally). The two
and Interleukin‑1‑beta (IL‑1 β), and proteolytic MESNA‑treated groups received MESNA (Uromitexan
enzymes such as matrix metalloproteinase (MMP), 75 mg/kg and 150 mg/kg, i.p.; Baxter, Germany),
leading to inflammation and degeneration of tissues.[3] respectively, 30 minutes before ischemia‑reperfusion.
It is well known that the acute inflammatory response The MESNA dose used in this study was modified based
leads to tissue damage through the production and on previous studies.[8,11,20]
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release of free radicals and cytotoxic proteins into

Carotid artery ischemia‑reperfusion procedure
the extracellular fluid.[4] Therefore, the need for an
anti‑inflammatory agent with immunomodulatory and The intraluminal thread method as described
by Handayani et al.[21] was used to induce
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free radical‑scavenging properties is increasing rapidly,

since inflammatory mediators play an increasingly vital ischemia‑reperfusion of the common carotid artery. The
role in ischemia‑reperfusion injury.[5] rats were placed supine, then the abdomen shaved and
cleaned with antiseptic solution (povidone iodine 10%).
2‑mercaptoethanesulfonate (MESNA) is a synthetic Abdominal incision was made 3 cm from the midline of
sulfur‑containing antioxidant and anti‑inflammatory drug the trachea, followed by exposure of the aorta and other
used to prevent nephrotoxic side effects associated with visceral arteries. The common carotid arteries were
chemotherapy drugs.[6] Studies have shown that MESNA exposed by midline blunt dissection of the sternohyoid
prevents ischemia‑reperfusion‑induced renal, intestinal, muscle. The common carotid artery was then ligated
and hepatic oxidative injury in rats,[4,7,8] burn‑induced bilaterally with 4/0 silk sutures at the level of the 4°–5°
renal injury in rats,[9] ulcerative colitis,[10] and several tracheal ring. The common carotid arteries were occluded
other inflammatory conditions.[11] MESNA exerts these for 10 min to induce ischemia and then subjected to 1 h
effects by regulating the activation of proteolytic enzymes of reperfusion. One hour after reperfusion, all animals
along with the expression of cytokines and suppression were decapitated, and blood and tissue samples of the
of free radicals.[4,12] Due to these cytoprotective effects, brain, heart, and lungs were collected.
MESNA may effectively limit the deleterious effects of
proinflammatory mediators and free radicals. Biochemical assays
Sera samples were centrifuged for 10 min at 1500 rpm and
Several experimental studies have shown therapeutics
stored at ‑80°C. To assess the severity of cellular injury,
using different agents to prevent ischemia‑reperfusion
serum levels of alanine transaminase (ALT), aspartate
injuries.[13‑17] However, several clinical trials of
aminotransferase (AST), alkaline phosphatase (ALP),
therapy to treat ischemia‑reperfusion injury have
and lactate dehydrogenase (LDH) were measured using
not yielded sufficient significant results,[18,19] and
an automated analyzer (BS240‑Vet, Mindray, Shenzhen,
there are also no studies on the effect of MESNA on
China). TNF‑α, IL‑1β, MMP(‑1, ‑2, ‑8), and TIMP‑1
carotid ischemia‑reperfusion. Therefore, based on an
serum levels were measured using rat‑specific ELISA
extensive literature search, we administered different
test kits (ELR‑TNF‑α and ELR‑IL1β, RaybioTech
doses of MESNA to investigate whether MESNA
Inc., GA, US). Assays were washed as described
confers protection against experimentally induced
by the manufacturer, using an automated microtiter
carotid ischemia‑reperfusion in rats by examining the
washer (MW‑12A Microplate Washer, Mindray,
histological changes in brain, heart, and lung tissues and
Shenzhen, China), and absorbances at 450 nm were
some serum biochemical parameters.
measured using a microtiter plate reader (MR‑96A
Materials and Methods Microplate Reader, Mindray, Shenzhen, China).
All experimental procedures used were approved Malondialdehyde (MDA), a stable lipid peroxidation
by the Near East University Ethics Committee with product, was measured in serum as previously described
protocol number 2021/139‑139. A total of 24 male by Beuge and Aust.[22] The assay was based on reaction
Wistar albino rats, about 3 months old and weighing with thiobarbituric acid (TBA) at 100°C in an acidic
250–300 g, were used for this study. All animals were environment, and the absorbance of the reaction mixture
anesthetized (100 mg/kg ketamine and 10 mg/kg was measured at 530–540 nm using a microplate
xylazine; i.p.) during all surgical procedures. reader (VersaMax Tunable Microplate Reader, Molecular
Devices LLC, CA, USA).
Experimental design
The rats were divided into four groups (n = 6) [Figure 1]. Histopathological assessment
In the sham‑operated control group, the common carotid After the decapitation of the animals, the tissue samples
artery was excised without occlusion. The carotid of brain, heart, and lungs were washed thoroughly with

942 Nigerian Journal of Clinical Practice ¦ Volume 26 ¦ Issue 7 ¦ July 2023

 Mercan, et al.: Effects of MESNA on carotid ischemia‑reperfusion in rats

saline. The samples were placed in 10% formaldehyde (75 and 150 mg/kg) (P < 0.05–0.01). However,
and routinely processed by embedding in paraffin. there was no significant difference between the
Tissue sections (5–6 mm) were stained with hematoxylin sham and either MESNA treatment groups, nor
and eosin and examined under a light microscope. An between the I/R + MESNA (75 and 150 mg/kg)
experienced histopathologist who was blinded to the groups (P > 0.05) [Table 1].
study design performed the histologic assessments. Effects of MESNA on serum malondialdehyde levels
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Statistical analysis The carotid I/R group showed a significant increase

Statistical analysis was performed using GraphPad Prism in serum MDA levels compared with the other
9.3.1 (GraphPad Software, San Diego, CA, USA). All groups (P < 0.001). MESNA (75 and 150 mg/kg)
data were expressed as mean ± (SEM). Datasets were significantly reversed the carotid I/R‑induced increase
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compared using one‑way analysis of variance (ANOVA) in serum MDA levels (P < 0.001). The I/R + MESNA
followed by Tukey’s post hoc test. Values ​​of P < 0.05 (75 and 150 mg/kg) groups showed no significant
were considered statistically significant. differences in MDA levels compared with the
sham group (P > 0.05). Serum MDA levels were
Results not significantly different between both MESNA
Biochemical results (75 and 150 mg/kg) groups (P > 0.05) [Table 2].
Effects of MESNA on serum enzyme activities Effects of MESNA on serum matrix metalloproteinases
Analysis of serum levels ALP, AST, ALT, and (MMP‑1, ‑2, ‑8) and tissue inhibitor of metalloprotease‑1
LDH levels in the carotid I/R group showed a (TIMP‑1) expression
highly significant increase compared with the sham, In the carotid I/R group, the serum levels of
I/R + MESNA (75 mg/kg), and I/R + MESNA MMP (‑1, ‑2, ‑8) and TIMP‑1 levels increased
(150 mg/kg) groups (P < 0.05–0.001). The carotid significantly compared to the other groups
I/R‑induced increase in serum enzyme levels (p < 0.05–0.0001). After treatment with MESNA
significantly decreased after treatment with MESNA (75 mg/kg and 150 mg/kg), serum levels of

Table 1: Serum levels of alkaline phosphatase (ALP), aspartate aminotransferase (AST), alanine transaminase (ALT),
and lactate dehydrogenase (LDH)
Sham Carotid I/R I/R + MESNA 75 mg/kg I/R + MESNA 150 mg/kg
ALP (U/L) 60.67±7.86 109±10.03*** 77.75±2.01# 68.37±6##
AST (U/L) 129±4.40 161.1±5.97** 129.8±6.69 ##
132.4±5.48##
ALT (U/L) 27.18±1.45 114.4±26.03** 35.35±7.40 ##
36.62±7.51##
LDH (U/L) 1360±153.7 2833±400.2** 1593±141.2 ##
1848±160.2#
Values are expressed as mean±standard error (n=6); **P<0.01, ***P<0.001 vs. sham; # P<0.05, ##P<0.01, vs. carotid I/R

Figure 1: Schematic diagram of the experimental design

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 Mercan, et al.: Effects of MESNA on carotid ischemia‑reperfusion in rats

Table 2: Serum malondialdehyde (MDA) levels

Sham Carotid I/R I/R + MESNA 75 mg/kg I/R + MESNA 150 mg/kg
MDA (µmol/L) 6.62±0.90 50.35±11.42*** 9.67±1.13### 7.42±1.12###
Values are expressed as mean±standard error (n=6); ***P<0.001 vs. sham; P<0.001 vs. carotid I/R
###

Table 3: Serum matrix metalloproteinases (MMP‑1, ‑2, ‑8) and tissue inhibitor of metalloprotease‑1 (TIMP‑1) levels
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Sham Carotid I/R I/R + MESNA 75 mg/kg I/R + MESNA 150 mg/kg
MMP‑1 (pg/mL) 1.51±0.15 3.82±0.19**** 2.49±0.28## 2.30±0.20###
MMP‑2 (pg/mL) 30.93±5.90 56.32±6.0** 28.69±2.27 ##
22.61±1.57###
MMP‑8 (pg/mL) 72.39±7.71 190.7±43.07** 96.48±4.75 #
82.79±4.74#
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TIMP‑1 (pg/mL) 383.7±45.13 899.2±71.42**** 640.2±69.94#* 405.6±62.83###

Values are expressed as mean±standard error (n=6); *P<0.05, **P<0.01, ****P<0.0001 vs. sham; #P<0.05, ##P<0.01, ###P<0.001 vs. carotid I/R

Table 4: Serum tumor necrosis factor‑alpha (TNF‑α) and Interleukin‑1‑beta (IL‑1β) levels
Sham Carotid I/R I/R + MESNA 75 mg/kg I/R + MESNA 150 mg/kg
TNF‑α (pg/mL) 13.36±3.10 34.17±6.17** 19.69±1.58# 8.59±1.61###
IL‑1β (pg/mL) 154.1±17.35 332.2±25.15**** 146.3±16.89 ####
113.4±15.79####
Values are expressed as mean±standard error (n=6); **P<0.01, ****P<0.0001 vs. sham; P<0.05, P<0.001, P<0.0001 vs. carotid I/R
# ### ####

Table 5: Effects of ischemia‑reperfusion and its treatment with MESNA on brain tissue
Sham Carotid I/R I/R + MESNA 75 mg/kg I/R + MESNA 150 mg/kg
Brain neutrophil layout 0.28±0.05 2.35±0.19**** 1.43±0.05****#### 1.23±0.04****####
Brain capillar intensity 0.25±0.02 2.4±0.07**** 1.7±0.05****####
1.23±0.04****####
Neuronal degeneration 0.33±0.05 2.9±0.07**** 2.55±0.04**** ##
2.26±0.06****####
Values are expressed as mean±standard error (n=6); ****P<0.0001 vs. sham; P<0.01, P<0.0001 vs. carotid I/R
## ####

and 150 mg/kg) groups in terms of MMP (‑1, ‑2,‑8),

and TIMP‑1 levels (P > 0.05) [Table 3].
Effects of MESNA on serum tumor necrosis
factor‑alpha (TNF‑α) and Interleukin‑1‑beta (IL‑1β)
expression
Analysis of serum TNF‑α and IL‑1β levels showed
a b a significant increase in the carotid I/R group
compared with the other groups (P < 0.05–0.0001).
The I/R + MESNA (75 and 150 mg/kg) groups
showed no significant differences compared with the
sham group (P > 0.05). MESNA (75 and 150 mg/kg)
significantly reversed the carotid I/R‑induced increase
in serum TNF‑α and IL‑1β levels (P < 0.05–0.0001).
c d There was no significant difference between the
Figure 2: Microscopic examination of brain tissues. (a) Sham group: I/R + MESNA (75 and 150 mg/kg) groups in terms of
regular neuropil morphology (*) and capillaries (arrows); (b) carotid TNF‑α and IL‑1β levels (P > 0.05) [Table 4].
I/R group: degenerated neurons (*) and capillaries (arrows); (c)
I/R + MESNA 75 mg/kg group: reduced neuronal degeneration (*) Histopathological results
capillaries (arrows); (d) I/R + MESNA 150 mg/kg group: regenerated
neuronal neurons (*) capillaries (arrows). (H.E, Bar 50 µm) Brain morphometric result
Brain neutrophil layout, brain capillary intensity,
MMP (‑1, ‑2,‑8) and TIMP‑1 decreased significantly and neuronal degeneration scores were significantly
compared to the carotid I/R group (p < 0.05–0.001); higher in the carotid I/R group than in the
however, there was significant increase in serum sham group (P < 0.0001). After treatment with
TIMP‑1 level after treatment with MESNA (75 mg/kg) MESNA (75 mg/kg and 150 mg/kg), the values of brain
compared with the sham group (P < 0.05). There was no histological features decreased significantly compared to
significant difference between the I/R + MESNA (75 the sham and carotid I/R groups (P < 0.0001). However,

944 Nigerian Journal of Clinical Practice ¦ Volume 26 ¦ Issue 7 ¦ July 2023

 Mercan, et al.: Effects of MESNA on carotid ischemia‑reperfusion in rats

Table 6: Effects of ischemia‑reperfusion and its treatment with MESNA on lung tissue
Sham Carotid I/R I/R + MESNA 75 mg/kg I/R + MESNA 150 mg/kg
Lung congestion 0.61±0.06 2.95±0.08**** 2.11±0.15****#### 1.78±0.08****####
Alveolar degeneration 0.51±0.06 2.86±0.18**** 2.21±0.14****#### 1.68±0.08****####++
Interstitial oedema 0.65±0.07 2.96±0.06**** 2.13±0.66****#### 1.61±0.13****####++
Values are expressed as mean±standard error (n=6); ****P<0.0001 vs. sham; ####P<0.0001 vs. carotid I/R; ++P<0.01: MESNA 75 vs.
MESNA 150 mg/kg
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Table 7: Effects of ischemia‑reperfusion and its treatment with MESNA on heart tissue
Sham Carotid I/R I/R + MESNA 75 mg/kg I/R + MESNA 150 mg/kg
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Heart congestion 0.5±0.08 2.8±0.03**** 2.3±0.07* 1.8±0.16#

Cardiomyocyte degeneration 0.3±0.03 2.51±0.04**** 2.08±0.06****### 1.78±0.06****####++
Inflammation 0.28±0.04 2.3±0.05**** 1.733±0.06* 1.4±0.07#
Values are expressed as mean±standard error (n=6); *P<0.05, ****P<0.0001 vs. sham; #P<0.05, ###P<0.001, ####P<0.0001 vs. carotid I/R;
++
P<0.01: MESNA 75 vs. MESNA 150 mg/kg

a b a b

c d c d
Figure 3: Microscopic examination of lung tissues. (a) Sham Figure 4: Microscopic examination of heart tissues. (a) sham group:
group: regular alveolar structure (arrows) and interstitial space, regular arrangement of cardiomyocytes and capillaries (*); (b) carotid
bronchiole (*); (b) carotid I/R group: severe and diffuse interstitial I/R group: severe congestion of the capillaries (arrow), cytoplasmic
edema (arrowhead) and capillary obstruction in hemorrhagic areas and disorganization in cardiomyocytes (*); (c) I/R + MESNA 75 mg/kg
decreased alveolar spaces (arrows); (c) I/R + MESNA 75 mg/kg group: group: reduced capillary congestion (arrowhead) and relative organization
improved alveolar structures (arrows) by regression of the interstitial of cardiomyocytes (*); (d) I/R + MESNA 150 mg/kg group: marked
edema (arrowhead) and bronchioles (*); (d) I/R + MESNA 150 mg/kg regression of capillary congestion (arrow) and regular pattern of
group: recovery highlighted in alveolar structure (arrows) and reduced cardiomyocytes (*). (H.E, Bar 50 µm)
interstitial edema (arrowheads). (H.E, Bar 50 µm)
Heart morphometric result
these changes caused by MESNA 75 and 150 mg/kg Heart congestion, cardiomyocyte degeneration, and
were not significantly different when both groups were inflammation scores were significantly higher in the
compared (P > 0.05) [Table 5]. carotid I/R group than in the sham group (P < 0.0001).
Lung morphometric result Treatment with MESNA especially high dose
Lung congestion, alveolar degeneration, and interstitial (150 mg/kg) decreased heart histological feature
edema scores were significantly higher in the carotid scores compared with the sham and carotid I/R
I/R group than in the sham group (P < 0.0001). groups (P < 0.05–0.0001). In addition, cardiomyocyte
Treatment with MESNA (75 mg/kg and 150 mg/kg) degeneration scores were significantly decreased more
significantly decreased the values of lung histological with MESNA 150 mg/kg than with MESNA 75 mg/kg
features compared with the sham and carotid I/R (P < 0.01) [Table 7].
groups (P < 0.0001). In addition, alveolar degeneration Microscopic presentation of lungs, heart, and brain
and interstitial edema scores were more significantly sections from the treated groups
decreased with MESNA 150 mg/kg than with MESNA Light microscopy examination of lung tissue in the
75 mg/kg (P < 0.01) [Table 6]. sham group showed a consistent alveolar structure

Nigerian Journal of Clinical Practice ¦ Volume 26 ¦ Issue 7 ¦ July 2023 945

 Mercan, et al.: Effects of MESNA on carotid ischemia‑reperfusion in rats

and interstitial space [Figure 3a]. In the carotid IR of proteolytic enzymes as well as proinflammatory
group, diffuse and severe hemorrhage with edema in cytokines, resulting in damage to brain, heart, and lung
interstitial spaces was observed. Besides, distended tissue.
alveolar walls and decreased alveolar space due to
Lipid peroxidation caused by oxidative degradation of
edema as well as severe leukocytes accumulation
polyunsaturated fatty acids in membranes is thought to
were observed [Figure 3b]. In IR + MESNA
play a key role in disrupting cell membrane integrity
75 mg/kg group, due to the regression of hemorrhage
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and cell lysis.[4] In the present study, MDA levels, a

in the interstitial space, an improvement in alveolar
product of lipid peroxidation,[24,25] were found to be
structure and a decrease in leukocyte accumulation
significantly elevated in serum as a result of carotid
were observed [Figure 3c]. In the I/R + MESNA
I/R. High MDA levels may indicate an increase in
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150 mg/kg group, the alveolar structure regenerated

lipid peroxidation, which can lead to oxidative tissue
besides regression of interstitial edema [Figure 3d].
injury due to intense free radical attack.[25,26] Our results
Light microscopy examination of heart tissue in showed that MESNA (75 and 150 mg/kg) significantly
the sham group showed a regular arrangement of decreased the I/R‑induced high serum MDA level to
cardiomyocytes and numerous capillaries [Figure 4a]. a comparable control level, as reported in previous
In the carotid IR group, remarkable interstitial edema studies.[4,8,27] Thus, MESNA may protect against carotid
and congestion in the capillaries and prominent ischemia‑reperfusion‑induced oxidative tissue damage
perinuclear and cytoplasmic disorganization in the by maintaining cell membrane integrity.
cardiomyocytes were observed [Figure 4b]. In the
In the current study, carotid I/R significantly altered
IR + MESNA 75 mg/kg group, the general cardiac
serum levels of AST, ALP, ALT, and LDH. Alteration
morphology showed a decrease in interstitial edema
in serum enzyme levels is indicative of organ
and congestion in capillaries and regeneration of
dysfunction.[8,25,28,29] MESNA (75 and 150 mg/kg)
cardiomyocytes [Figure 4c]. In the I/R + MESNA
significantly reversed the carotid I/R‑induced increase
150 mg/kg group, congestion was markedly reduced and
in serum enzyme levels. This decrease in serum enzyme
regular cardiomyocytes were prominent [Figure 4d].
levels after MESNA treatment could be attributed
Light microscopy examination of brain tissue in the to the ability of MESNA to prevent the leakage
sham group revealed regular morphology of neurons of intracellular enzymes from damaged cells by
and neuropils in the cortex [Figure 2a]. However, the promoting cell regeneration and tissue repair. Our
carotid I/R group [Figure 2b] showed degenerated results are consistent with previous studies on the
neurons with marked shrinkage of the nucleus and protective effects of intraperitoneally administered
cytoplasm. The I/R + MESNA 75 mg/kg group showed MESNA.[30,31]
less neuronal degeneration compared with the carotid
The role of proinflammatory cytokines in 1/R‑induced
I/R group [Figure 2c]. The I/R + MESNA 150 mg/kg
tissue injury has been described in several previous
group showed regeneration of neuropil morphology
studies.[5,8,32] In this study, we investigated the effect of
[Figure 2d].
MESNA on the serum concentration of TNF‑α and IL‑1β,
Discussion which are cytokines with different biological functions,
especially in inflammatory and immune responses. These
Occlusion of the carotid artery triggers cerebral ischemia, proinflammatory cytokines serve as systemic markers of
causing significant local tissue damage. The heart and tissue injury.[33] In our study, carotid artery I/R triggered
lungs are two distant organs that are susceptible to the a significant increase in serum levels of TNF‑α and
adverse effects of carotid ischemia‑reperfusion.[1,23]
IL‑1β, which was consistent with the results of previous
Therefore, the aim of the present study was to investigate
studies that reported that carotid artery I/R increased
the effects of MESNA on local and distant organ
serum levels of proinflammatory cytokines.[5,32] Increased
damage caused by carotid ischemia‑reperfusion (I/R) by
serum levels of TNF‑α and IL‑1β indicate infiltration of
assessing the proinflammatory cytokines and proteolytic
inflammatory cells into the injured tissue and increased
enzyme pathways.
cytokine production, leading to local exacerbation of
The carotid artery is occluded in experimental studies injury and spread of inflammation to distant organs.[34]
to induce ischemia because it is a widely accepted MESNA (75 and 150 mg/kg) reversed the I/R‑induced
experimental model for ischemia‑reperfusion that increase in serum TNF‑α and IL‑1β, and histopathological
mimics the characteristics of acute cerebrovascular results revealed that tissue damage was significantly
injury in humans.[1,21] The current data show that prevented. These results are consistent with the previous
carotid I/R causes a significant increase in serum levels studies.[31,35] The ability of MESNA to regulate the

946 Nigerian Journal of Clinical Practice ¦ Volume 26 ¦ Issue 7 ¦ July 2023

 Mercan, et al.: Effects of MESNA on carotid ischemia‑reperfusion in rats

inflammatory process in ischemia‑reperfusion could be Financial support and sponsorship

through several mechanisms, including downregulation Nil.
of nuclear factor‑κB (NF‑κB) activity,[36,37] inhibition
Conflicts of interest
of myeloperoxidase and inducible nitric oxide,[36,38]
and monocyte chemotactic protein‑1, which regulates There are no conflicts of interest.
the migration and infiltration of proinflammatory
proteins.[39,40]
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