AVE0991 Attenuates Pyroptosis And Liver Damage After Heatstroke By Inhibiting The ROS-NLRP3 Inflammatory Signalling Pathway

BackgroundLiver damage,a lethal complication after heatstroke,has rarely been elucidated in terms of its exact mechanism.Our two previous studies demonstrated that the liver damage after heatstroke associated with pyroptosis induced by ROS-NLRP3 inflammatory signalling pathway.Not only that,the rennin-angiotensin system(RAS)played a role in regulating ROS-NLRP3 pathways in hepatocytes.ObjectiveThe objective of this study was to explore not only the changing rule of RAS(renin-angiotensin system)of patients and rats after heatstroke for the first time,but also the possibility that intervention with RAS alleviated the liver damage by inhibiting ROS-NLRP3 inflammatory signalling pathway.Methods1.The changing rule of RAS in the serum of heatstroke patients with liver damageThe serum samples and the medical records of heatstroke patients were collected.The ROC curve method and others were used to analyze the predictive value of RAS as a potential biomarker when predicting liver damage.2.The establishment of a conscious rat model after heatstroke monitored arterial blood pressureThe conscious rat model after heatstroke monitored arterial blood pressure was constructed and compared with anesthetized rat model in blood pressure,body temperature,heat load,fluid loss,and survival rate.The influence of intervention agent of RAS was also explored.3.The changing rule and influence of RAS on the heatstroke ratsThe changing rule of RAS components in the serum and liver tissues of rats after heatstroke were analyzed.Their relationship with liver damage and reactive oxygen species(ROS)was also evaluated.4.AVE 0991 attenuating pyroptosis and liver damage after heatstroke by inhibiting the ROS-NLRP3 inflammatory signalling pathwayIn vitro and vivo,the expression levels of NOX4,NLRP3,caspase 1,1L-1βprotein in hepatocytes induced by heat stress were detected.The effect of AVE 0991 on the ROS-NLRP3 pathway as a protective agent was investigated.Results1.In the serum of heatstroke patients with liver damage,higher serum level of angiotensin(Ang)Ⅱ and lower level of Ang-(1-7)were noticed.Ang Ⅱ,Ang-(1-7)and Ang Ⅱ/Ang-(1-7)ratio displayed significant qualification when predicting the occurrence of liver damage.2.For the conscious rat model after heatstroke monitored arterial blood pressure,the gentler changing rules of body temperature and blood pressure,more fluid loss,higher thermal load and lower short-term mortality rate were observed.3.There was an increase in Ang Ⅱ and a decrease in Ang-(1-7)sampling from the serum and liver tissue of rats after heatstroke.This imbalance was accompanied by increased damage and ROS levels in the liver.4.In vitro and vivo,Ang Ⅱ promoted the damage and increased the ROS level of the hepatocytes stimulated by heat stess.AVE 0991 improved that and inhibited the up-regulation of NOX4,NLRP3,caspase-1and 1L-1β protein levels.Conclusion1 Ang Ⅱ and Ang-(1-7)were potential biomarkers when predicting the liver damage after heatstroke.2.The concious rat model after heatstroke monitored arterial blood pressure was more appropriate as pathological physiology state,especially in assessing hemodynamics.3.The imbalance of RAS and increased level of ROS were demonstrated for the rats after heatstroke,which was relieved using RAS intervention.4.AVE 0991 attenuated pyroptosis and liver damage after heatstroke by inhibiting the ROS-NLRP3 inflammatory signalling pathway.