Explore The Mechanism And Clinical Efficacy Of Zhibitai On NAFLD Based On The MAPK-ERK-TLRs Pathway

Research Background Nonalcoholic Fatty Liver Disease(NAFLD)is a metabolic stress liver damage closely related to Insulin Resistance and genetic susceptibility.The disease spectrum includes Nonalcoholic Hepatic Steatosis,Nonalcoholic Steatohepatitis(NASH),and NASH-related Cirrhosis and Hepatocellular Carcinoma.NAFLD is a new challenge in the contemporary medical field,and its harm to human health will continue to expand.In the meantime,there are no clinically marketed drugs for NAFLD at home and abroad.The modernization of Chinese Medicine inspires us to make full use of our country's existing rich resources of Chinese Medicine.It has become a top priority for us to choose to develop original medicinal materials or original prescriptions reasonably and find effective drugs that can not only improve Insulin Resistance but also protect the liver and anti-inflammatory.This research consists of two parts: the basic research part and the clinical research part.The basic research part used high-fat and high-sugar to induce Non-alcoholic Fatty Liver Disease on animal models to explore the effect of Zhibitai intervention on NAFLD and its influence on the MAPK-ERK-TLRs signaling pathway.This aimed to provide an experimental basis for Zhibitai's clinical treatment for NAFLD.The clinical research part used Zhibitai to treat NAFLD patients and observed the effects of Zhibitai on the Liver Stiffness Index(LSM)and Fat Attenuation Index(CAP),and the improvement of other indicators including Liver Function,Blood Lipids,Blood Sugar,and Insulin Resistance Index(HOMA-IR),and comprehensive evaluation of Zhibitai's therapeutic effect on NAFLD patients.The basic research partObjective: To establish a rat model of Non-alcoholic Fatty Liver Disease,and observe the effect and mechanism of Zhibitai in the treatment of Don-alcoholic Fatty Liver Disease in diseased animal models.Methods: In the experiment,6-week-old male SD rats were selected,and 10% fructose and high-fat diet were used to induce a Non-alcoholic Fatty Liver Disease rat model with IR.After the models were successfully verified,the model rats were randomly divided into a Model Control Group,Zhibitai Low-dose Group(100 mg/kg/day),Zhibitai High-dose Group(200 mg/kg/day),Metformin Group(200 mg/kg/day),Silybin Group(100 mg/kg/day),and Normal Control Group.During the administration period,the animal body weight changes were monitored.After 9 weeks of continuous administration,blood samples were collected from the rats.A Blood Glucose Meter was used to check Fasting Blood Glucose(FBG)and 2 hours Postprandial Blood Glucose(2h BG)of the rats in each group.The Insulin Resistance Index(HOMA-IR)was calculated.The Automatic Blood Biochemical Analyzer was used to check the levels of Serum Alanine Aminotransferase(ALT),Aspartate Aminotransferase(AST),Total Cholesterol(CHOL),Triglycerides(TG),High-density Lipoprotein Cholesterol(HDL-C),and Low-density Lipoprotein Cholesterol(LDL-C).The ELISA kit was used to check the levels of Serum Hyaluronidase(HA),Laminin(LN),IV Collagen(IV-C),and Type III Procollagen(PC-?).The weight of the liver and the Liver Index were calculated.HE staining was used to observe the pathological changes of liver tissue.The oil red O staining was used to observe the deposition of lipid droplets in frozen sections of liver tissue;The Masson staining was used to observe liver tissue fibrosis degree.And the q PCR and Western bolt were used to check the levels of TLR4 and P-ERK1/2 m RNA,and protein in liver tissue.Results: Compared with the Normal Control Group,2h BG,Fins,HOMA-IR,ALT,AST,TG,CHOL,HDL-C,LDL-C,HA,LN,IV-C,PC-?,Liver Index and Pathology Scores of the Model Groups were significantly increased while HDL-C was significantly reduced.TLR4,P-ERK1/2 m RNA,and protein expression levels were significantly increased.Compared with the Model Control Groups,the 2h BG,Fins,HOMA-IR,ALT,AST,TG,CHOL,LDL-C,Liver Index,and Histopathological Scores of Zhibitai low and high dose groups were significantly reduced,and HDL-C and AST/ ALT were significantly increased,and TLR4,P-ERK1/2 m RNA and protein expression levels were significantly reduced.Both the high and low doses groups of Zhibitai could down-regulate the expression of serum HA,LN,IV-C,PC-?,and collagen fibers in liver tissues of model animals.Conclusion: Zhibitai can prevent the occurrence and development of inflammation,improve Blood Lipids,and antagonize Insulin Resistance,improve the vacuolar degeneration of the liver,reduce the inflammation of the liver,and protect the liver by down-regulating the expression of TLR4 and P-ERK1/2 proteins and affecting the MAPK-ERK-TLRs pathway to inhibit the transduction of inflammatory signal pathways.It can significantly reduce the level of serum fibrosis markers and the level of collagen fibers in liver tissue and has a therapeutic effect on non-alcoholic fatty liver disease.The Clinical research partObjective: To comprehensively evaluate the clinical therapeutic effect of Zhibitai on patients with Non-alcoholic Fatty Liver Disease and provide a reference for the clinical application of Zhibitai.Methods: The experiment adopted a prospective research method.In the past year,75 NAFLD patients with phlegm and blood stasis were the research objects.The patients were randomly divided into 2 groups: 37 in the Control Group and 38 in the Experimental Group.9 patients fell off during the test,and the actual number of effective cases was 66,including 33 cases in the Experimental Group and 33 cases in the Control Group.Patients in the Control Group only controlled their diet and strengthened exercise.Patients in the Experimental Group were given Zhibitai capsules on the basis of diet control and strengthened exercise.1 capsule(0.24g)each time,twice a day,12 weeks as a course of treatment.Observe the liver CAP value,LSM value,the weight,BMI,Liver Function(ALT,AST,GGT,ALP),Blood Lipids(TC,TG,LDL-C,HDL-C),Blood Sugar(GLU,Ins,Hb A1c),Resistance Index(HOMA-IR)of before and after treatment improvement situation of the two groups.Clinical trials used liver CAP,LSM,the weight and BMI as the main efficacy indicators and Liver Function,Blood Lipids,Blood Sugar,and HOMA-IR as secondary efficacy indicators for evaluation.Results: Compared with before-treatment,the weight,BMI,LSM,and CAP of the Control Group decreased statistically(P<0.05)after treatment.There was no significant change in other test indicators.The weight,BMI,LSM,CAP,ALT,AST,GGT,GLU,Ins,HOMA-IR,CHOL,and LDL-C of the Experimental Group after treatment had a statistically significant decrease(P<0.05),and there were no significant changes in other test indicators.Compared with the Control Group before treatment,there was no statistical difference in the test indicators of the Experimental Group before treatment.Compared with the Control Group after treatment,the LSM,CAP,ALT,AST,GGT,GLU,Ins,HOMA-IR,CHOL and LDL-C of the Experimental Group had a statistically significant decrease after treatment.There was no statistical difference in other test indicators.Conclusion: Basic treatment for 12 weeks can significantly reduce the weight,BMI,LSM,and CAP of the patients with the Non-alcoholic Fatty Liver to achieve the main therapeutic effect,but there is no significant improvement in Liver Function,Blood Lipid regulation,and Insulin Resistance.Zhibitai intervention plus basic treatment for 12 weeks can significantly reduce the weight,BMI,LSM,and CAP of the patients with the Non-alcoholic Fatty Liver to achieve the main therapeutic effect.At the same time,it can significantly improve the Liver Function,Lower Blood Lipids,and improve Insulin Resistance.The treatment effect of the Experimental Group with Non-alcoholic fatty Liver Disease is better than that of the Control Group.In addition,Zhibitai can significantly improve Liver Function,Blood Lipids,and Insulin Resistance.Zhibitai treatment for Non-alcoholic fatty Liver patients reached the primary and secondary treatment endpoints.