Experimental Protocol for Cecal Ligation and Puncture Model of Polymicrobial Sepsis and Assessment of Vascular Functions in Mice

Sepsis is the systemic inflammatory response syndrome that occurs during infection and is exacerbated by the inappropriate immune response encountered by the affected individual. Despite extensive research, sepsis in humans is one of the biggest challenges for clinicians. The high mortality rate in sepsis is primarily due to hypoperfusion-induced multiorgan dysfunctions , resulting from a marked decrease in peripheral resistance. Vascular dysfunctions are further aggravated by sepsis-induced impairment in myocardial contractility. Circulatory failure in sepsis is characterized by refractory hypotension and vascular hyporeactivity (vasoplegia) to clinically used vasoconstrictors. To investigate the complex pathophysiology of sepsis and its associated multiple organ dysfunction, several animal models have been developed. However, cecal ligation and puncture (CLP) model of murine sepsis is still considered as 'gold standard' in sepsis research. In this protocol we have described the standard surgical procedure to induce polymicrobial sepsis by cecal ligation and puncture. Further, we have described the protocol to study the molecular mechanisms underlying vascular dysfunctions in sepsis.