Summary

Cecal Ligation पंचर प्रक्रिया

Published: May 07, 2011
doi:

Summary

cecal और मानव पूति के अध्ययन के लिए एक महत्वपूर्ण उपकरण के रूप में ligation पंचर के माउस मॉडल.

Abstract

Human sepsis is characterized by a set of systemic reactions in response to intensive and massive infection that failed to be locally contained by the host. Currently, sepsis ranks among the top ten causes of mortality in the USA intensive care units 1. During sepsis there are two established haemodynamic phases that may overlap. The initial phase (hyperdynamic) is defined as a massive production of proinflammatory cytokines and reactive oxygen species by macrophages and neutrophils that affects vascular permeability (leading to hypotension), cardiac function and induces metabolic changes culminating in tissue necrosis and organ failure. Consequently, the most common cause of mortality is acute kidney injury. The second phase (hypodynamic) is an anti-inflammatory process involving altered monocyte antigen presentation, decreased lymphocyte proliferation and function and increased apoptosis. This state known as immunosuppression or immune depression sharply increases the risk of nocosomial infections and ultimately, death. The mechanisms of these pathophysiological processes are not well characterized. Because both phases of sepsis may cause irreversible and irreparable damage, it is essential to determine the immunological and physiological status of the patient. This is the main reason why many therapeutic drugs have failed. The same drug given at different stages of sepsis may be therapeutic or otherwise harmful or have no effect 2,3. To understand sepsis at various levels it is crucial to have a suitable and comprehensive animal model that reproduces the clinical course of the disease. It is important to characterize the pathophysiological mechanisms occurring during sepsis and control the model conditions for testing potential therapeutic agents.

To study the etiology of human sepsis researchers have developed different animal models. The most widely used clinical model is cecal ligation and puncture (CLP). The CLP model consists of the perforation of the cecum allowing the release of fecal material into the peritoneal cavity to generate an exacerbated immune response induced by polymicrobial infection. This model fulfills the human condition that is clinically relevant. As in humans, mice that undergo CLP with fluid resuscitation show the first (early) hyperdynamic phase that in time progresses to the second (late) hypodynamic phase. In addition, the cytokine profile is similar to that seen in human sepsis where there is increased lymphocyte apoptosis (reviewed in 4,5). Due to the multiple and overlapping mechanisms involved in sepsis, researchers need a suitable sepsis model of controlled severity in order to obtain consistent and reproducible results.

Protocol

1. Cecal और मानव पूति के लिए एक माउस मॉडल के रूप में Ligation पंचर / 6 चूहों (7-9 पुराने सप्ताह) इस प्रक्रिया C57BL के लिए उपयोग किया जाता है. Intraperitoneally 01:01 (75mg/kg) ketamine और xylazine (15mg/kg) के एक समाधान इंजेक्शन द्वारा माउस anesthetize. …

Discussion

यहाँ हम विस्तार से बताते हैं कि कैसे चूहों में CLP मॉडल का प्रदर्शन करने के लिए और गंभीरता के ग्रेड मिलाना.

पूति के अन्य पशु मॉडल की तुलना में, CLP विभिन्न आयु और लिंग के किसी भी माउस तनाव में प्रदर…

Divulgazioni

The authors have nothing to disclose.

Acknowledgements

इस काम के स्वास्थ्य के पेंसिल्वेनिया विभाग से अनुदान द्वारा समर्थित किया गया था.
डॉ. Miguel Garcia Toscano मंदिर विश्वविद्यालय में एक postdoctoral साथी, इस अध्ययन के दौरान Alfonso मार्टिन Escudero फाउंडेशन द्वारा वित्त पोषित किया गया था.
हम वीडियो के बनाने के लिए Iliya Yordanov और केविन Kotredes शुक्रिया अदा करना चाहता हूँ.

Materials

Name of the reagent Company Catalogue number Comments
Ketamine Ketaset 0856-2013-01  
Xylazine AnaSed NADA# 139-236  
Insulin syringe 29G Exel 26028  
Silk suture, 6-0 PROLENE Ethicon 8680G  
19G and 25G Needle BD 305186  
Shaver General supply General supply  
Infrared Heating lamp General supply General supply  
Michel wound clips 7mm Roboz Surgical Instr RS-9270  
Ear Loop Mask Fisher Scientific 19-130-4181  
Dissection scissors Roboz Surgical Instr RS-6702  
Betadine solution VWR 63410-992  
Surgical forceps Roboz Surgical Instr RS-5135  
70% Isopropyl alcohol pad Fisher Scientific 22-031-350  
Buprenorphine Bedford Labs 55390-100-10  
Tramadol Sigma-Aldrich 42965  

Riferimenti

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Citazione di questo articolo
Toscano, M. G., Ganea, D., Gamero, A. M. Cecal Ligation Puncture Procedure. J. Vis. Exp. (51), e2860, doi:10.3791/2860 (2011).

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