To Investigate The Clinical Efficacy And Mechanism Of Jiawei-lizhong Decoction In The Treatment Of Non-alcoholic Steatohepatitis Based On PI3K/AKT/mTOR Signaling Pathway

Objective:Through clinical observation,the clinical efficacy and safety of Jiawei-Lizhong decoction in the treatment of non-alcoholic steatohepatitis were fully explored,and the intervention effect of Jiawei-Lizhong decoction on related serum indexes in NASH rats was further explored by using PI3K/AKT/mTOR signaling pathway as a breakthrough point.To detect the expression of proteins in the PI3K/AKT/mTOR pathway and key proteins of autophagy in rats,and to explore whether its therapeutic mechanism is to activate autophagy by inhibiting PI3K/AKT/mTOR signaling pathway,so as to provide modern scientific basis for the treatment of NASH with Jiawei-Lizhong decoction.Methods:(1)Clinical study:64 patients with NASH who met the inclusion criteria were randomly divided into treatment group and control group,32 cases in each group.The treatment group was treated with Jiawei-Lizhong decoction,and the control group was treated with vitamin E soft capsules.Both groups were treated for 12 weeks based on the correction of bad life behavior.The changes of TCM symptom score,liver function,blood lipid and liver transient elasticity test before and after treatment were observed and compared between the two groups,and statistical analysis and efficacy evaluation were performed.(2)Experimental study:After one week of adaptive feeding,34 SPF SD rats were randomly divided into 6 rats as blank group,and the remaining 28 rats were fed with high-fat diet for 12 weeks.After 12 weeks,4 rats were randomly selected to verify the success of modeling,and the remaining 24 rats were numbered independently and then randomly divided into model group,high dose Chinese medicine group,medium dose Chinese medicine group and low dose Chinese medicine group by SPSS 23.0 software,with 6 rats in each group.After that,the rats in the TCM high dose group were given gavage of Wei-Lizhong Decoction(2×14.38g/kg/d),the rats in the TCM medium dose group were given gavage of Wei-Lizhong decoction(14.38g/kg/d),and the rats in the low dose group were given gavage of Wei-Lizhong decoction(0.5×14.38g/kg/d).The model and blank groups were given the same dose of distilled water by gavage.The treatment lasted for 4 weeks.Rats in each group were sacrificed at the end of 16 weeks,and blood samples and liver tissue samples were collected.Liver function(ALT,AST)and blood lipid(TC,TG,HDL-C,LDL-C)were detected.The pathological morphology of rat liver tissue was observed by high power microscope.The protein expression levels of p-PI3K,p-AKT,p-mTOR,Beclinl,P62,and LC3Ⅱ were detected by Western Blot.Results:(1)Clinical research:during the observation process,4 cases were excluded due to loss of follow-up,non-compliance with the requirements of treatment and other reasons,and the actual effective cases were 60 cases.①Clinical symptom score:the total effective rate of the treatment group and the control group(Chinese medicine group,western medicine group)were 93.3%and 76.7%,respectively.The total effective rate of the treatment group was better than that of the control group(P<0.05).②Comparison of liver function:there were significant differences before and after treatment in each group(P<0.01).The levels of ALT and AST in the treatment group were lower than those in the control group after treatment(P<0.05).③Blood lipid:TG,TC,HDL-C and LDL-C were significantly different between the two groups before and after treatment(P<0.01).After treatment,the blood lipids(TG,TC,HDL-C,LDL-C)in the treatment group were significantly improved compared with those in the control group,and the difference was statistically significant(P<0.05).④ Comparison of liver stiffness and fat attenuation:the liver stiffness and fat attenuation of each group after treatment were significantly improved compared with those before treatment,and the difference was statistically significant(P<0.01).The liver stiffness of the treatment group after treatment was significantly lower than that of the control group(P<0.05),and the fat attenuation of the treatment group after treatment was significantly better than that of the control group(P<0.05).⑤TCM symptom score:after treatment,the TCM symptom score of the treatment group was significantly improved than that of the control group(P<0.01).⑥Safety evaluation:There were no adverse reactions and toxic side effects in this clinical experiment,and there were no abnormal safety indicators.(2)Animal experiment:After the intervention of traditional Chinese medicine,the nutritional status,mental state and activity of rats in each treatment group were improved compared with those before treatment.①Liver function and blood lipid indexes:compared with the blank group,the levels of ALT,AST,TC,TG and LDL-C in the model group increased significantly(P<0.01),while HDL-C decreased significantly.Compared with the model group,the levels of ALT,AST,TC,TG and LDL-C in each TCM treatment group decreased to varying degrees,while HDL-C increased(P<0.01),and the high-dose TCM treatment group had the most obvious effect.②Expression of p-PI3K,p-AKT,p-mTOR,P62,Beclinl and LC3II in liver tissue of rats in each group:Compared with the blank group,the expression of p-PI3K,p-Akt,p-mTOR and P62 in liver tissue of the model group was significantly up-regulated,and the expression of Beclinl and LC3II was decreased(P<0.01).Compared with the model group,the expressions of p-PI3K,p-Akt,p-mTOR and P62 in the Chinese medicine treatment groups were decreased to varying degrees,and the expressions of Beclinl and LC3II were increased to varying degrees(P<0.05).The improvement of protein expression in the high dose group was better than that in the middle and low dose groups(P<0.05,P<0.01).③Liver tissue pathology:the liver tissue pathology of the model group showed extensive enlargement and swelling of hepatocytes,nuclear deviation,large vesicular changes,"balloon" changes,cell cord disorder,inflammatory cell infiltration in the portal area,scattered hepatocyte spots and focal necrosis.Compared with the model group,the inflammatory infiltration and the number of fat vacuoles were significantly reduced in the TCM treatment groups,and the high-dose TCM treatment group had the most significant improvement.Conclusion:(1)Jiawei-Lizhong Decoction can relieve the clinical symptoms of NASH patients,improve the liver function,blood lipids,liver stiffness and liver fat attenuation degree of patients,and there is no obvious adverse event in the process of the study,which is more advantageous than vitamin E soft capsule in the treatment of NSAH.(2)Jiawei-Lizhong Decoction can effectively improve the general condition,liver function,blood lipid level,and lipid metabolism of NASH rats,and the expressions of p-PI3K,p-AKT,p-mTOR and P62 proteins in the treatment groups were down-regulated to varying degrees,and the expressions of Beclinl and LC3II were increased to varying degrees.These results suggest that autophagy may be activated by inhibiting the PI3K/AKT/mTOR pathway.