To Explore The Mechanism Of Sini Decoction On LPS-induced Septic Shock Rats Based On The Heat-sensitive Channel TRPV1

Background:Background:Sini Decoction originated from Treatise on Febrile Diseases,the ancestor of medical formulary.Formulas of Chinese Medicine depicted it as the interior-warming prescription,restoring yang and rescuing patient from collapse.It consists of three herbs:monkshood,rhizoma zingiberis and honey-fried licorice root.This prescription cures yang depletion and mistreatment of perspiration by Shaoyin disease and Taiyang disease.In modern times,Sini Decoction is used to cure all kinds of shock,cerebral infarction,myocardial infarction,cardiac failure,sepsis and sick sinus syndrome."Monkshood generates no heat without ginger".The combination of monkshood and rhizoma zingiberis is quite important in Sini Decoction.On this basis,the research aims to discuss the efficacy of Sini Decoction.Modern studies indicate thermo-sensitive channels are important factors for septic shock.TRPV1 channel has the protective effects.According to existing studies,Sini Decoction contains different kinds of TRPV1 agonists.TRPV activation can alleviate symptoms by septic shock.Septic shock is regarded as an emergent and severe disease,which is concerned with multiple organ failures and micro-circulation disturbances.TRPV1 can regulate internal environment of human body.In this research,physical signs,biochemical criterion measurement,immunohistochemistry and immunity protein technologies are adopted,so as to analyze the function of thermo-sensitive channel TRPV1 in Sini Decoction curing septic shock.Research purpose:Mechanism of Sini Decoction curing septic shock.Method of research:1.Animal experiment:①grouping:normal group,model group,Sini Decoction group,positive control group(capsaicin is a positive control drug);②dosage:PBS PBS 1Omg/kg,LPS 10mg/kg,capsaicin 1mg/kg,Sini Decoction 2.7g/kg(Daily dose of crude drug);③time:blood pressure,heart rate and other indicators were measured before LPS administration,1h,5h,and 24h after administration;④biochemical indicator test:determination of indicators related to septic shock in rat blood.2.Immunohistochemical method and immunoblotting are used to conduct semi-quantitative observation on TRPV1 and downstream proteins CGRP,NK1R in the hearts and kidneys of rats.Results:1.Results of animal experimentPhysical signs of animal experiments shows that,after 1 hour of LPS injection,the heart rate of the model group,Sini Decoction group and the positive control group was significantly higher than that of the normal group(P<0.05,P<0.001,P<0.01).After LPS injection for 5h to 24h,the heart rate of the model group,Sini Decoction group and positive control group was significantly increased compared with the normal group(P<0.001).After 24 hours of LPS injection,the heart rate of Sini Decoction group and positive control group was significantly lower than that of the model group(P<0.01).After 5 hours of LPS injection,the blood pressure of the model group decreased significantly compared with the normal group(P<0.01).Compared with the model group,the blood pressure of the positive control group was significantly increased(P<0.05).After 24 hours of LPS injection,the blood pressure of the model group and the Sini Decoction group decreased significantly compared with the normal group(P<0.001,P<0.01).Compared with the model group,the blood pressure of the Sini Decoction group and the positive control group increased significantly(P<0.01,P<0.001).Mortality results in animal experiments showed that the mortality of the model group and the Sini Decoction group increased significantly(P<α’)at 24 h after LPS injection compared with the normal group.Compared with the model group,there was no significant difference in mortality between the Sini Decoction group and the positive control group(P>a’).The results of biochemical indicators showed that the levels of TNFα,IL-6 and IL-1β in the model group were significantly higher than those in the normal group(P<0.001).Compared with the model group,the contents of TNFα,IL-6 and IL-1β in the Sini Decoction group and the positive control group were significantly decreased(P<0.001).Compared with the normal group,the plasma levels of NE in the model group and Sini Decoction group were significantly increased(P<0.001,P<0.05).Compared with the model group,the plasma levels of NE in the Sini Decoction group and the positive control group were significantly decreased(P<0.01,P<0.001).2.Results of molecular biology experimentalThe results of immunohistochemistry showed that there was no significant difference between the model group and the treatment group(P>0.05).After 24 hours of LPS injection in rats,the expression of TRPV1,CGRP and NK1R in the kidney of the model group was significantly up-regulated compared with the normal group(P<0.01).Compared with the model group,the expressions of TRPV1,CGRP and NK1R in Sini Decoction group and positive control group were significantly down-regulated(P<0.05).The results of immunoblotting showed that compared with the normal group,the expression of TRPV1 in the kidney of the model group and the positive control group was significantly up-regulated(P<0.001,P<0.05).Compared with the model group,the expression of TRPV1 was significantly down-regulated in the Sini Decoction group and the positive control group(P<0.001).Compared with the normal group,the expression of CGRP in the kidney of the model group was significantly up-regulated(P<0.001).Compared with the model group,the expression of CGRP in the Sini Decoction group and the positive control group was significantly down-regulated(P<0.001).Compared with the normal group,the expression of NK1R in the kidney of the model group was significantly up-regulated(P<0.001).Compared with the model group,the expression of NK1R was significantly down-regulated in the Sini Decoction group and the positive control group(P<0.01,P<0.001).Conclusion:Desensitization of TRPV1 is one of the important molecular mechanisms for Sini Decoction to have protective effects on septic shock.