SIMULATION OF EXPERIMENTAL SEPSIS BY CECAL LIGATION AND PUNCTURE (CLP)

Author(s):  
E.Yu. Shapovalova ◽  
G.A. Demyashkin ◽  
M.Yu. Malanichev ◽  
D.A. Pogosyan ◽  
I.A. Zorin ◽  
...  

Currently, the problem of treating sepsis is acute. To study these morphological and functional changes, animal models are used, for example, a model of experimental peritonitis, cecal ligation and puncture (CLP). However, there is only insufficient research on the description of internal organ rearrangements, in particular, skin morphological picture. The aim of the study was to assess of changes in mice internal organs in case of sepsis modeling. Materials and Methods. The authors performed cecal ligation and puncture in mice (n=40) to form experimental peritonitis and severe sepsis. In the control group (n=10), a sham surgery was conducted: a midline laparotomy was followed by layer-by-layer deaf suturing of the surgical wound. Results. The authors observed CLP-induced disorders in all vital organs, especially in the liver (violation of the beam structure of the hepatic lobules with signs of balloon dystrophy and necrosis areas, leukocyte infiltration, plethora of sinusoids), kidneys (thinning of the visceral layer of the Bowman’s capsule, narrowing of the afferent arteriole lumen, balloon dystrophy of proximal and distal tubules, widespread disappearance of the brush border in nephrocytes) and the spleen (hyperplasia of the white pulp with a large number of apoptotic lymphocytes). Moreover, signs of mild inflammatory infiltration were observed in the skin. Conclusion. The morphological changes found during the study corresponded to the reaction of the test organs in sepsis. The proposed CLP method for experimental peritonitis can be used as a sepsis model. Keywords: sepsis, cecal ligation and puncture (CLP), skin, inflammation. В настоящий момент остро стоит проблема лечения сепсиса. Для изучения данных морфофункциональных изменений используют модели на животных, например модель экспериментального перитонита – лигирование и пункцию слепой кишки (cecal ligation and puncture, CLP). Однако исследований, касающихся описания перестроек внутренних органов, в частности морфологической картины кожного покрова, проведено явно недостаточно. Цель исследования. Морфологическая оценка изменений внутренних органов мышей в условиях моделирования сепсиса. Материалы и методы. У мышей (n=40) проводили лигирование и пункцию слепой кишки для формирования экспериментального перитонита (CLP) и тяжелого сепсиса. В контрольной группе (n=10) осуществляли «фиктивную» операцию – срединную лапаротомию с последующим послойным глухим ушиванием операционной раны. Результаты. Во всех жизненно важных органах наблюдали нарушения, индуцированные CLP, особенно в печени (нарушение балочного строения печеночных долек с признаками баллонной дистрофии и зонами некроза, лейкоцитарная инфильтрация, полнокровие синусоидов), почках (истончение висцерального листка капсулы Боумена–Шумлянского, сужение просвета приносящих артериол, баллонная дистрофия проксимальных и дистальных канальцев, повсеместное исчезновение щеточной каемки в нефроцитах) и селезенке (гиперплазия белой пульпы с наличием большого количества апоптотических лимфоцитов), а также отмечали признаки слабой воспалительной инфильтрации в коже. Заключение. Обнаруженные в ходе исследования морфологические изменения соответствуют реакции исследуемых органов при сепсисе. Предложенный метод CLP для создания экспериментального перитонита можно использовать в качестве модели сепсиса. Ключевые слова: сепсис, cecal ligation and puncture (CLP), кожа, воспаление.

Circulation ◽  
2018 ◽  
Vol 138 (Suppl_2) ◽  
Author(s):  
Jing Xu ◽  
Guanghui Zheng ◽  
Liangliang Wu ◽  
Xiangshao Fang ◽  
Yue Wang ◽  
...  

Introduction: Abnormal levels of end-tidal carbon dioxide (ETCO 2 ) may reflect a derangement in perfusion, metabolism, or gas exchange. It is unclear if ETCO 2 can be used for fluid resuscitation (FR) compared with traditional mean arterial pressure (MAP) as an outcome predictor in sepsis. Hypothesis: Use of ETCO2 is better than MAP in guiding fluid resuscitation to improve lactate levels and microcirculatory blood flow in sepsis. Methods: Thirty-five male Sprague-Dawley rats weighing 350-400g were randomized to: 1) SHAM, n=5; 2) cecal ligation and puncture (CLP) Control group (with CLP, without FR, n=10); 3) ETCO 2 group (with CLP, FR began when ETCO 2 ≤25 mmHg, n=10) and 4) MAP group (with CLP, FR began when MAP≤100 mmHg, n=10). Lactate level, cardiac output (CO), perfused small vessel density (PSVD) and sublingual microvascular flow index (MFI) was assessed at baseline, 2 h, 4 h, 8 h, 10 h and 12 h post-CLP. Survival duration was recorded. Results: After FR,CO in the ETCO 2 group increased compared with the MAP group 12h after CLP while lactate levels decreased compared with the Control and MAP groups (p<0.05) (Figure-1). Both sublingual PSVD and MFI decreased after CLP in the control group and MAP group but significantly improved in the ETCO 2 group 8h post-CLP. The average survival time in the ETCO 2 group was significantly greater than MAP group (Figure-2). Conclusions: ETCO 2 guided FR was associated with improved CO, lactate, microcirculatory flow, and survival time compared to MAP guided FR in a CLP-induced rat model of sepsis.


2020 ◽  
Vol 2020 ◽  
pp. 1-9
Author(s):  
Qian-wei Li ◽  
Qin Yang ◽  
Hong-Yang Liu ◽  
Yu-ling Wu ◽  
Yu-Hua Hao ◽  
...  

Sepsis increases the risk of the liver injury development. According to the research works, coenzyme Q10 exhibits hepatoprotective properties in vivo as well as in vitro. Current work aimed at investigating the protective impacts of coenzyme Q10 against liver injury in septic BALB/c mice. The male BALB/c mice were randomly segregated into 4 groups: the control group, the coenzyme Q10 treatment group, the puncture and cecal ligation group, and the coenzyme Q10+cecal ligation and puncture group. Cecal ligation and puncture was conducted after gavagaging the mice with coenzyme Q10 during two weeks. Following 48 h postcecal ligation and puncture, we estimated hepatic biochemical parameters and histopathological changes in hepatic tissue. We evaluated the expression of factors associated with autophagy, pyroptosis, and inflammation. Findings indicated that coenzyme Q10 decreased the plasma levels in alkaline phosphatase, alanine aminotransferase, and aspartate aminotransferase in the cecal ligation and puncture group. Coenzyme Q10 significantly inhibited the elevation of sequestosome-1, interleukin-1β, oligomerization domain-like receptor 3 and nucleotide-binding, interleukin-6, and tumor necrosis factor-α expression levels; coenzyme Q10 also increased beclin 1 levels. Coenzyme Q10 might be a significant agent in the treatment of liver injury induced by sepsis.


1987 ◽  
Vol 253 (2) ◽  
pp. E130-E134
Author(s):  
J. A. Spitzer ◽  
I. V. Deaciuc

Inositol trisphosphate-dependent Ca2+ release was measured in saponin-permeabilized hepatocytes isolated from acutely (2 mg/100 g body wt iv) or chronically (0.1 mg X 100 g body wt-1 X 24 h-1 for 30 h) endotoxin-treated (ET, Escherichia coli) rats or from animals rendered septic by cecal ligation and puncture. A decrease of this parameter was observed in acutely ET-treated rats (52%, P less than 0.01) and after 30 h of continuous ET infusion (33%, P less than 0.01). Sepsis was associated with an elevated Ca2+ release (34%, P less than 0.01) as compared with the sham-operated animals. We conclude that during endotoxicosis and sepsis alterations of intracellular Ca homeostasis take place, reaching sites beyond the level of the plasma membrane. Such alterations could account in part for metabolic and functional changes associated with these pathologic states. In addition, ET treatment provides the first known intervention resulting in the modulation of inositol trisphosphate-dependent Ca2+ release.


2021 ◽  
Vol 18 (6) ◽  
pp. 1161-1166
Author(s):  
Zhiwen Zhou ◽  
Xiang Ren ◽  
Aiping Li ◽  
Wensheng Zhou ◽  
Li Huang

Purpose: To investigate the protective effect of floroindole against cecal ligation and puncture (CLP)- induced sepsis, as well as the underlying mechanism of action. Methods: Thirty-five 10–week-old male Wistar rats weighing 190 - 210 g (mean: 200.00 ± 10.10 g) were used for this study. The rats were randomly assigned to seven groups of five rats each, viz, normal control group, and six CLP groups. The CLP groups were those subjected to cecal ligation and puncture (CLP). The first 5 CLP groups received 2, 4, 6, 8 or 10 mg/kg floroindole, respectively, 1 h after CLP, via intraperitoneal route (i.p.) while the 6th CLP group served as untreated control. Western blotting, enzyme-linked immunosorbent assay (ELISA) and real-time quantitative polymerase chain reaction (qRT-PCR) were used for the assessment of the expression levels of tumor necrosis factor-α (TNF- α), interleukn-6 (IL-6), nucleotide-binding oligomerization domain 2 (NOD2) and p-NF-κB p65. Results: Cecal ligation and puncture (CLP) significantly and time-dependently upregulated the expressions of TNF-α, IL-6 and NOD2 in intestinal tissues of rats (p < 0.05). However, treatment with floroindole significantly, and dose-dependently down-regulated CLP-induced expressions of these proteins (p < 0.05). Treatment of rats with floroindole also significantly and dose-dependently inhibited CLP-induced phosphorylation of NF-κB p65 in rat ileum (p < 0.05). Conclusion: The results obtained in this study demonstrate that floroindole confers some degree of protection against CLP-induced sepsis via inhibition of NF-κB p65 phosphorylation.


2018 ◽  
Vol 2018 ◽  
pp. 1-6 ◽  
Author(s):  
Sebastián P. Chapela ◽  
Isabel Burgos ◽  
Christian Congost ◽  
Romina Canzonieri ◽  
Alexis Muryan ◽  
...  

In sepsis, reactive oxygen species (ROS) production is increased. This process takes place mainly within the electron transport chain. ROS production is part of the pathophysiology of multiple organ failure in sepsis. Succinate yields Dihydroflavine-Adenine Dinucleotide (FADH2), which enters the chain through complex II, avoiding complex I, through which electrons are lost. The aim of this work is to determine if parenteral succinate reduces systemic ROS production and improves kidney function. Rats with cecal ligation and puncture were used as model of sepsis, and 4 groups were made: Control group; Succinate group, which only received parenteral succinate; Sepsis group; and Sepsis which received parenteral succinate. Systemic ROS are measured 24 hours after the procedure. Rats subjected to cecal puncture treated with succinate had less systemic ROS than Septic untreated rats (p=0.007), while there were no differences in creatinine levels (p=0.07). There was no correlation between creatinine and systemic ROS levels (p=0.3). We concluded that parenteral succinate reduces ROS levels, but it does not reduce creatinine levels. Since there is no correlation between both levels, the processes would not be related.


2008 ◽  
Vol 4 (1) ◽  
pp. 31-36 ◽  
Author(s):  
Daniel Rittirsch ◽  
Markus S Huber-Lang ◽  
Michael A Flierl ◽  
Peter A Ward

PLoS ONE ◽  
2013 ◽  
Vol 8 (10) ◽  
pp. e77936 ◽  
Author(s):  
Shao-Chun Wu ◽  
Johnson Chia-Shen Yang ◽  
Cheng-Shyuan Rau ◽  
Yi-Chun Chen ◽  
Tsu-Hsiang Lu ◽  
...  

2018 ◽  
Vol 7 (2) ◽  
pp. 90-94
Author(s):  
E. I. Shurygina ◽  
V. S. Polyakova ◽  
V. A. Mikhanov

The aim of the research is to study the dynamics of structural and functional changes in the thyroid and parathyroid glands during reparative osteogenesis. Material and methods. The study was performed on 40 adult males rat of the Wistar line, weighing 180.0±10.0 g. A model of an open fracture of the tibial diaphysis was used in the experimental group. The control group was intact. Structural changes in bone callus, thyroid and parathyroid glands were studied using immunohistochemistry methods, light microscopy, morphometry and statistics. Results. The proliferative and functional activity of the major endocrinocytes of the parathyroid glands is increased in the initial periods of reparative osteogenesis, resulting in an increasing of osteoclastic activity with resorption of bone fragments in the fracture zone. From 21th day of osteogenesis the functional activity of the parathyroid glands is reduced, the activity of the C-cells of the thyroid gland is activated; then the functional activity of C-cells is decreased. Conclusion. The observed changes in the synthetic and proliferative activity of C-cells and parathyrocytes have a certain time interval, which correlates with the histogenesis of bone tissue in the fracture zone.


2021 ◽  
Vol 99 (2) ◽  
pp. 185-191
Author(s):  
Sonja Salinger-Martinovic ◽  
Vladan Cosic ◽  
Nenad Stojiljkovic ◽  
Sonja Ilic ◽  
Nikola Stojanovic ◽  
...  

Doxorubicin is an anticancer agent that is commonly used to treat a number of tumors and is associated with acute and chronic changes of the cardiovascular system. Ellagic acid has strong free radical scavenging capacity, neuroprotective and hepatoprotective effects, and is known to protect against changes occurring due to diabetes, cardiovascular diseases, and cancer. Twenty-four Wistar rats were divided in four groups: control group received saline, doxorubicin group received doxorubicin in a single dose of 20 mg/kg, ellagic acid group received ellagic acid in a dose of 4 mg/kg, and doxorubicin + ellagic acid group received doxorubicin and ellagic acid in same doses as in previous groups. The effect of ellagic acid treatment, alone or in combination with doxorubicin, was studied on isolated heart frequency and strength of the contraction, and on thoracic aorta contractile responses. Application of ellagic acid to rats pre-treated with doxorubicin significantly prevented functional changes occurring in the heart, but not in the thoracic aorta tissue. Ellagic acid statistically significantly (p < 0.001) prevented doxorubicin-induced increase in heart rate, while at the same time increased single contraction force (p < 0.001) and attenuated morphological changes on heart tissue induced by doxorubicin. We can conclude that ellagic acid has potential to prevent doxorubicin-induced changes of the cardiovascular system.


PRILOZI ◽  
2015 ◽  
Vol 36 (3) ◽  
pp. 13-25 ◽  
Author(s):  
Elida Mitevska ◽  
Irena Kostadinova-Petrova ◽  
Nevena Kostovska

AbstractThe aim of our investigation was to evaluate the immunosuppressive effect of medroxyprogesterone acetate (MPA) determining the volume densities of the structural components of the spleen. The volume densities of the same structural components of spleen were determined after administration of dexamethasone too, in order to see whether the morphological changes induced by MPA are in the same line with the changes caused by dexamethasone.60 female Wistar rats were divided into 5 groups. The control group of rats was administered physiological solution. The remaining, 4 experimental groups were administered: dexamethasone at a therapeutic daily dose of 0.6 mg/kg bw and maximal therapeutic dose of 3 mg/kg bw, and MPA at a therapeutic dose of 30 mg/kg bw and maximal therapeutic dose of 150 mg/kg bw. The drugs were applied intramuscularly for 7 days. Spleen paraffin sections were stained according to the methods: hematoxylin-eosin, Masson and Elastica van-Gieson. Stereological measurements were performed by using the Weibl′s multipurpose test system (M-42).The histological analyses of the structural components of the spleen in rats treated with dexamethasone and MPA have shown reduction of the white pulp and the marginal zone and an apparent decrease of the cellular density of the lymphocyte component of the pulp. The stereological analysis of the spleen showed significant decrease of the splenic pulp volume density and significant increase of the connective tissue volume density. Reducing the presence of splenic pulp was mainly due to the decrease in the volume density of all structural components of the white pulp. Changes were observed in all drug treated groups of rats.Our results have shown that the MPA provoked changes suggested atrophy of the spleen lymphoid tissue. Although the atrophic changes of the spleen were significant after the application of both dexamethasone and MPA, the white pulp was significantly more sensitive substrate for dexamethasone than for the MPA.


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