Study On The Protective Effects Of MCC950 And VX765 On Ketamine-induced Liver Injury In Rats

Ketamine is an anesthetic drug widely used in human and veterinary clinics.It is usually used in different indications such as pediatric patients and infant animal surgical anesthesia.However,during physical development,exposure to anesthetics may cause serious side effects on the growth and development and nervous system of infants and young animals.Ketamine is extensively metabolized by microsomal enzymes in the liver into metabolites,which can damage liver parenchymal cells and other liver cells.In recent years,a variety of animal model diseases in vitro and in vivo have gradually become the focus of research through pathological mechanisms such as pyroptosis.In the pyroptosis pathway,the most important component is the NLRP3 inflammasome,which is also the most widely studied and most characteristic inflammasome.The activation of inflammasomes promotes the release of inflammatory factors and causes pyroptosis.The hepatotoxicity caused by ketamine anesthesia may be related to the pathway of pyroptosis.MCC950 and VX765 act as inhibitors of two pyroptosis pathways.Among them,MCC950 can inhibit the activation of NLRP3,and VX765 can effectively inhibit the process of pyrosis by inhibiting Caspase-1.However,whether these two inhibitors can protect liver cell pyroptosis induced by ketamine is still less studied and the mechanism is still unclear.Therefore,this experiment is divided into two parts.First,7-day-old rats are selected to be anesthetized with ketamine to explore the possible liver toxicity.After determining the liver injury,MCC950 and VX765 two inhibitors were used to explore whether they can protect young rats from the hepatotoxicity caused by ketamine anesthesia.So as to reduce the impact of anesthetics on young animals,provide new therapeutic targets,and provide theoretical support for relatively safe anesthesia methods in the future.In the first part of the experiment,7-day-old rats were randomly divided into 2 groups,the control group(group C)and the ketamine group(group K).The rats in group C were the normal saline control group.The rats in group K were intraperitoneally injected with ketamine(dose 20mg/kg,injected once every 90 minutes to maintain the young rats under anesthesia.Ketamine was injected 5 times for a total of 7.5 hours.);In the second part of the experiment,young rats were randomly divided into 6 groups,the C and K groups are the same as the first part,the MCC950 inhibitor group(M group: 10 mg/kg)and the VX765 inhibitor group(V group: 25 mg/kg)was also administered by intraperitoneal injection,and the ketamine + MCC950 inhibitor group(KM group)and the ketamine+VX765 inhibitor group(KV group)were injected with two drugs each.The inhibitor was administered half an hour in advance after preparation,and the administration method was the same as that of group K.After the anesthesia is over,the rat liver tissue and blood are collected.Evaluation of liver damage based on changes in liver function indicators,expression of inflammatory factors and HE staining;immunohistochemical staining method was used to detect the protein expression of key pyroptosis proteins NLRP3,ASC,Caspase-1 and GSDMD;q RT-PCR method was used The m RNA expression of pyroptosis-related genes NLRP3,ASC,Caspase-1 and GSDMD was detected;Western blot was used to detect the protein expression changes of NLRP3,ASC,Caspase-1 and GSDMD in the signal pathway.Results:(1)In this test,ketamine caused obvious lesions in the liver tissues of group K,the arrangement of liver cells was disordered,the liver cells appeared swelling,and multiple inflammatory infiltrations of hepatocytes could be observed.However,after the addition of MCC950 and VX765 inhibitors,it was able to Significantly alleviate the degree of liver damage.The swelling of hepatocytes is reduced,and the hepatocytes are still arranged in a cord shape,with a small amount of inflammatory infiltration.(2)The level of serum inflammatory factors in group K showed an increasing trend(P<0.01).After adding the inhibitor,KM and KV group showed a decreasing trend compared with K group(P<0.01),M and V group had no significant difference compared with C group(P>0.05).(3)Ketamine anesthesia increased the content of MDA in the liver of young rats,and decreased the content of GSH and SOD activity.Pretreatment with inhibitors of MCC950 and VX765 could alleviate the degree of oxidative stress.(4)The experimental results show that prolonged anesthesia or exposure to ketamine leads to an increase in the expression of pyroptosis-related proteins in young rats.The expression of NLRP3,ASC,Caspase-1,and GSDMD protein in the liver tissue of 7-day-old rats in the K group increased(P<0.01);the relative K group protein expression in the KM and KV groups decreased(P< 0.05)(5)Compared with group C,NLRP3,ASC,Caspase-1 and GSDMD m RNA in group K were significantly increased(P<0.01).However,there was a significant downward trend in the KM group and the KV group(P<0.01).The results show that ketamine can cause liver damage in young rats by inducing cell pyroptosis,and MCC950 and VX765 inhibitors can effectively reduce the liver damage caused by exposure to ketamine in young rats and have a protective effect.