Profiling Circulating MicroRNA Expression in Experimental Sepsis Using Cecal Ligation and Puncture

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.

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Immunodesign of experimental sepsis by cecal ligation and puncture

Nature Protocols ◽

10.1038/nprot.2008.214 ◽

2008 ◽

Vol 4 (1) ◽

pp. 31-36 ◽

Cited By ~ 835

Author(s):

Daniel Rittirsch ◽

Markus S Huber-Lang ◽

Michael A Flierl ◽

Peter A Ward

Keyword(s):

Cecal Ligation ◽

Experimental Sepsis ◽

Cecal Ligation And Puncture

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Sophorolipids block lethal effects of septic shock in rats in a cecal ligation and puncture model of experimental sepsis*

Critical Care Medicine ◽

10.1097/01.ccm.0000196212.56885.50 ◽

2006 ◽

Vol 34 (1) ◽

pp. E188 ◽

Cited By ~ 53

Author(s):

Martin H. Bluth ◽

Emad Kandil ◽

Catherine M. Mueller ◽

Vishal Shah ◽

Yin-Yao Lin ◽

...

Keyword(s):

Septic Shock ◽

Cecal Ligation ◽

Experimental Sepsis ◽

Cecal Ligation And Puncture ◽

Lethal Effects ◽

Puncture Model

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Autonomic dysfunction in experimental sepsis induced by cecal ligation and puncture

Autonomic Neuroscience ◽

10.1016/j.autneu.2007.10.006 ◽

2008 ◽

Vol 138 (1-2) ◽

pp. 57-63 ◽

Cited By ~ 33

Author(s):

João Alexandre Trés Pancoto ◽

Pollyanna Barbosa Farias Corrêa ◽

Gabriela Ravanelli Oliveira-Pelegrin ◽

Maria José Alves Rocha

Keyword(s):

Autonomic Dysfunction ◽

Cecal Ligation ◽

Experimental Sepsis ◽

Cecal Ligation And Puncture

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A model-specific role of microRNA-223 as a mediator of kidney injury during experimental sepsis

AJP Renal Physiology ◽

10.1152/ajprenal.00493.2016 ◽

2017 ◽

Vol 313 (2) ◽

pp. F553-F559 ◽

Cited By ~ 18

Author(s):

James F. Colbert ◽

Joshay A. Ford ◽

Sarah M. Haeger ◽

Yimu Yang ◽

Kyrie L. Dailey ◽

...

Keyword(s):

Cecal Ligation ◽

Kidney Injury ◽

Organ Injury ◽

Multiple Models ◽

Infection Model ◽

Experimental Sepsis ◽

Bacterial Clearance ◽

Cecal Ligation And Puncture ◽

Slurry Injection

Sepsis outcomes are heavily dependent on the development of septic organ injury, but no interventions exist to interrupt or reverse this process. microRNA-223 (miR-223) is known to be involved in both inflammatory gene regulation and host-pathogen interactions key to the pathogenesis of sepsis. The goal of this study was to determine the role of miR-223 as a mediator of septic kidney injury. Using miR-223 knockout mice and multiple models of experimental sepsis, we found that miR-223 differentially influences acute kidney injury (AKI) based on the model used. In the absence of miR-223, mice demonstrated exaggerated AKI in sterile models of sepsis (LPS injection) and attenuated AKI in a live-infection model of sepsis (cecal ligation and puncture). We demonstrated that miR-223 expression is induced in kidney homogenate after cecal ligation and puncture, but not after LPS or fecal slurry injection. We investigated additional potential mechanistic explanations including differences in peritoneal bacterial clearance and host stool virulence. Our findings highlight the complex role of miR-223 in the pathogenesis of septic kidney injury, as well as the importance of differences in experimental sepsis models and their consequent translational applicability.

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Von Willebrand Factor-Dependent Inflammatory Responses in Mouse Septic Model By Cecal Ligation and Puncture

Blood ◽

10.1182/blood.v124.21.2773.2773 ◽

2014 ◽

Vol 124 (21) ◽

pp. 2773-2773

Author(s):

Shogo Kasuda ◽

Hideto Matsui ◽

Shiro Ono ◽

Yasunori Matsunari ◽

Kenji Nishio ◽

...

Keyword(s):

Survival Rate ◽

Mouse Model ◽

Von Willebrand Factor ◽

Inflammatory Responses ◽

Cecal Ligation ◽

Experimental Sepsis ◽

Bolus Administration ◽

Cecal Ligation And Puncture ◽

Von Willebrand ◽

Willebrand Factor

Abstract Sepsis is a serious inflammatory response syndrome, in which systemic activation of both inflammatory and coagulation pathways are provoked by severe microbial infection. In addition to the known hemostatic functions, von Willebrand factor (VWF) is recently assumed to participate in a cross-talk between inflammation and thrombosis. Indeed, recent mouse model studies demonstrated that proper functional regulation of VWF-dependent inflammatory responses by ADAMTS13 considerably improved the disease progression of brain stroke or myocardial infarction. However, little is known about the detailed mechanisms or relevant role of VWF functions in inflammation. We therefore studied the physiologic relevance of VWF-dependent inflammatory responses in a mouse model of experimental sepsis by cecal ligation and puncture (CLP). The mouse CLP was performed according to the established standard protocol (Rittirsch D, et al., Nat Protoc, 2009). Briefly, mouse cecum was ligated distal to the ileocecal valve under laparotomy, punctured with 18 gauge needle and gently pressed until a small drop of stool appeared. The cecum was returned to the peritoneal cavity and 200 μL of saline was injected into the cavity to avoid dehydration before body wall and skin incision were closed with a 4-0 Sofsilk. We compared 17 wild-type (WT) and 17 VWF-gene deleted (knock-out; KO) mice (from The Jackson Laboratory, Bar Harbor, ME), all of which were 10-12 weeks of age, healthy and fertile. Excess blood loss was not observed in all (WT or KO) mice during the CLP operation. Kaplan-Meier analysis revealed the significantly (p < 0.05) lower survival rate of KO mice than that of WT (KO; 23.5% vs. WT; 58.8% at the Day 7 of CLP). The impaired survival rate of KO mice was restored by the bolus administration of human VWF (n=21, 2.5 U/mouse) to an extent comparable to that of WT. Peripheral blood analysis of KO mice at 24 hours after CLP showed the severe leukocytopenia with sharp decrease of neutrophils in blood, as compared to WT. In addition, formation of walled-off abscess was confirmed at the peri-cecal space in all the WT and KO mice alive even at the Day 7 of CLP, while such focal abscess formation was not found in mice died before the Day 3 of CLP. Thus, our results altogether indicate that VWF could play a role on the recruitment or accumulation of neutrophils for microbial killing at the local inflammatory focus. VWF-mediated platelet aggregate formation in peripheral capillary vessels then could shut down the local microcirculation, thereby blocking systemic microbial expansion as a crucial biological defense mechanism. Disclosures Shima: Chugai Pharmaceutical Co., Ltd.: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding.

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Effects of dexamethasone and adrenocorticotropin on circulating microRNA expression in humans

Endocrine Abstracts ◽

10.1530/endoabs.37.oc9.1 ◽

2015 ◽

Author(s):

Ivan Igaz ◽

Gabor Nyiro ◽

Zoltan Nagy ◽

Pal Perge ◽

Miklos Toth ◽

...

Keyword(s):

Microrna Expression ◽

Circulating Microrna

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Juglone Attenuates Sepsis-Induced Acute Lung Injury via Suppression of the TLR4/Nf-κb Pathway

Current Topics in Nutraceutical Research ◽

10.37290/ctnr2641-452x.18:201-206 ◽

2020 ◽

Vol 18 (2) ◽

pp. 201-206

Author(s):

Qiu Nan ◽

Xu Xinmei ◽

He Yingying ◽

Fan Chengfen

Keyword(s):

Acute Lung Injury ◽

Lung Injury ◽

Nuclear Factor Kappa B ◽

Cecal Ligation ◽

Myeloperoxidase Activity ◽

Nuclear Factor ◽

Cecal Ligation And Puncture ◽

Toll Like Receptor ◽

Toll Like Receptor 4 ◽

Nuclear Factor Kappa

Sepsis, with high mortality, induces deleterious organ dysfunction and acute lung injury. Natural compounds show protective effect against sepsis-induced acute lung injury. Juglone, a natural naphthoquinone, demonstrates pharmacological actions as a pro-apoptotic substrate in tumor treatment and anti-inflammation substrate in organ injury. In this study, the influence of juglone on sepsis-induced acute lung injury was investigated. First, a septic mice model was established via cecal ligation and puncture, and then verified via histopathological analysis of lung tissues, the wet/dry mass ratio and myeloperoxidase activity was determined. Cecal ligation and puncture could induce acute lung injury in septic mice, as demonstrated by alveolar damage and increase of wet/dry mass ratio and myeloperoxidase activity. However, intragastric administration juglone attenuated cecal ligation and puncture-induced acute lung injury. Secondly, cecal ligation and puncture-induced increase of inflammatory cells in bronchoalveolar lavage fluid was also alleviated by the administration of juglone. Similarly, the protective effect of juglone against cecal ligation and puncture-induced acute lung injury was accompanied by a reduction of pro-inflammatory factor secretion in bronchoalveolar lavage fluid and lung tissues. Cecal ligation and puncture could activate toll-like receptor 4/nuclear factor-kappa B signaling pathway, and administration of juglone suppressed toll-like receptor 4/nuclear factor-kappa B activation. In conclusion, juglone attenuated cecal ligation and puncture-induced lung damage and inflammatory response through inactivation of toll-like receptor 4/nuclear factor-kappa B, suggesting a potential therapeutic strategy in the treatment of sepsis-induced acute lung injury.

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Inactivation of Nf-κb Pathway by Taxifolin Attenuates Sepsis-Induced Acute Lung Injury

Current Topics in Nutraceutical Research ◽

10.37290/ctnr2641-452x.18:176-182 ◽

2019 ◽

Vol 18 (2) ◽

pp. 176-182

Author(s):

Chen Weiyan ◽

Deng Wujian ◽

Chen Songwei

Keyword(s):

Acute Lung Injury ◽

B Cells ◽

Lung Injury ◽

Signaling Pathway ◽

Light Chain ◽

Cecal Ligation ◽

Nuclear Factor ◽

Kappa Light Chain ◽

Cecal Ligation And Puncture ◽

Light Chain Enhancer

Acute lung injury is a clinical syndrome consisting of a wide range of acute hypoxemic respiratory failure disorders. Sepsis is a serious complication caused by an excessive immune response to pathogen-induced infections, which has become a major predisposing factor for acute lung injury. Taxifolin is a natural flavonoid that shows diverse therapeutic benefits in inflammation- and oxidative stress-related diseases. In this study, we investigated the role of taxifolin in a mouse model of cecal ligation and puncture-induced sepsis. Cecal ligation and puncture-operated mice presented damaged alveolar structures, thickened alveolar walls, edematous septa, and hemorrhage compared to sham-treated controls. Cecal ligation and puncture mice also showed increased wet-to-dry (W/D) lung weight ratio and elevated total protein concentration and lactate dehydrogenase level in bronchoalveolar lavage fluid. Taxifolin treatment protected animals against sepsis-induced pulmonary damage and edema. Septic mice presented compromised antioxidant capacity, whereas the administration of taxifolin prior to cecal ligation and puncture surgery decreased malondialdehyde concentration and enhanced the levels of reduced glutathione and superoxide dismutase in mice with sepsis-induced acute lung injury. Moreover, cecal ligation and puncture-operated mice showed markedly higher levels of proinflammatory cytokines relative to sham-operated group, while taxifolin treatment effectively mitigated sepsis-induced inflammation in mouse lungs. Further investigation revealed that taxifolin suppressed the activation of the nuclear factor kappa-light-chain-enhancer of activated B cells signaling pathway in cecal ligation and puncture-challenged mice by regulating the phosphorylation of p65 and IκBα. In conclusion, our study showed that taxifolin alleviated sepsis-induced acute lung injury via the inhibition of nuclear factor kappa-light-chain-enhancer of activated B cells signaling pathway, suggesting the therapeutic potential of taxifolin in the treatment sepsis-induced acute lung injury.

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P717Glucagon-like peptide 1 (GLP-1) improves endothelial dysfunction and vascular inflammation in polymicrobial sepsis induced by cecal ligation and puncture (CLP)

European Heart Journal ◽

10.1093/eurheartj/ehz747.0322 ◽

2019 ◽

Vol 40 (Supplement_1) ◽

Author(s):

S Steven ◽

J Helmstaedter ◽

F Pawelke ◽

K Filippou ◽

K Frenies ◽

...

Keyword(s):

Oxidative Stress ◽

Clinical Trials ◽

Endothelial Dysfunction ◽

Knockout Mice ◽

Vascular Inflammation ◽

Cecal Ligation ◽

Cecal Ligation And Puncture ◽

Glucose Levels ◽

Dpp4 Inhibitor ◽

Cardioprotective Effects

Abstract Objective Sepsis causes severe hypotension, accompanied by high mortality in the setting of septic shock. LEADER, SUSTAIN-6 and other clinical trials revealed cardioprotective and anti-inflammatory properties of GLP-1 analogs like Liraglutide (Lira). We already demonstrated improved survival by amelioration of disseminated intravasal coagulation (DIC) in lipopolysaccharide (LPS)-induced endotoxemia by inhibition of the GLP-1 degrading enzyme dipeptidylpeptidase-4 (DPP-4). With the present study we aim to investigate the mechanism of protective effects of the GLP-1 analog Lira and the DPP4 inhibitor Linagliptin (Lina) in the clinically relevant sepsis model cecal ligation and puncture (CLP). Methods C57/BL6j and endothelial cell-specific GLP-1 receptor knockout mice (Cdh5crexGLP-1rfl/flmice) were used and sepsis was induced by cecal ligation and puncture (CLP). DPP4 inhibitor (Lina, 5mg/kg/d; 3 days) and GLP-1 analog (Lira, 200μg/kg/d; 3 days) were applied subcutaneously. Aortic vascular function was tested by isometric tension recording. Aorta and heart tissue was used for Western blotting, dot blot and qRT-PCR. Endogenous GLP-1 (7–36 and 9–36) and insulin was determined by ELISA. Blood samples were collected for examination of cell count, oxidative stress and glucose levels. Results Body temperature was increased by CLP and normalized by Lina and Lira. Sham- and Lira- but not Lina-treated septic mice showed low blood glucose levels compared to healthy controls. Acetylcholine-induced (endothelium-dependent) vascular relaxation in aorta was impaired by CLP. This was accompanied by vascular inflammation and elevation of IL-6, iNOS, ICAM-1, and TNF-alpha mRNA levels in aortic tissue. Vascular, cardiac and whole blood oxidative stress were increased by CLP. Furthermore, we detected higher levels of IL-6, 3-nitrotyrosine (3-NT) and 4-hydroxynonenal (4-NHE) in plasma of CLP animals. Lina and Lira reduced oxidative stress and vascular inflammation, which was accompanied by improved endothelial function. In addition, CLP treatment in endothelial specific knockout mice of the GLP-1r strongly induced mortality compared to WT mice, with the effect being strongest in the Lira-treated group. Conclusion The present study demonstrates that Lina (DPP4 inhibitor) and the GLP-1 analog Lira ameliorate sepsis-induced endothelial dysfunction by reduction of vascular inflammation and oxidative stress. Clinical trials like LEADER and SUSTAIN-6 proved that GLP-1 analogs like Lira have cardioprotective effects in T2DM patients. The present study, performed in a clinically relevant model of polymicrobial sepsis, reveals that the known cardioprotective effects of GLP-1 might be translated to other diseases which affect the cardiovascular system like sepsis, underlining the potent anti-inflammatory effects of GLP-1 analogs.

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