Study On Mechanism Of Yinqi Tiaozhi Decoction In Treating NAFLD By Regulating LXRα Pathway

Objective:The study aims at observeing the clinical efficacy of Yinqi Tiaozhi Decoction in the treatment of non-alcoholic fatty liver disease(NAFLD)and clarifying its mechanism of traditional Chinese medicine through animal experiments in order to provide evidence for clinical use.Methods:Theoretical research: By reviewing ancient and modern literature,we summarized the relevant research of traditional Chinese and western medicine,including the historical origin,epidemiology and treatment of NAFLD.We also discussed the status and role of LXRα pathway and related targets in NAFLD.Based on the theory of "liver stagnation and turbidity generation",the etiology and pathogenesis of NAFLD was probed into.Under the guidance of this theory,the method of clearing liver and removing turbidity was taken as the treatment principle and the prescription of Yinqi Tiaozhi Yin was discussed.Clinical research: 73 patients with Damp-heat accumulation syndrome were randomly divided into treatment group and control group.There were 3 cases of shedding,finally 35 cases in the treatment group and 35 cases in the control group.Both groups were given health education and guidance to change their life style.On this basis,patients in the treatment group took Yinqi Tiaozhi Decoction orally and patients in the control group took silybin capsule orally.After 3 months,the changes of serum ALT,AST,GGT,ALP,TG,TC,HDL-C,LDL-C,FFA levels,TCM syndromes scores,liver ultrasound image,CAP values and the incidence of adverse reactions were observed before and after treatment in order to evaluate the efficacy and safety.Experimental Study: NAFLD rats were induced by high-fat diet(HFD).50 male SD rats were randomly divided into 9 normal groups and 41 model groups.Rats in normal group were given normal diet,rats in model group were given high-fat diet.After 12 weeks,we randomly chose one rat from each of the two groups and stained for pathological sections of liver tissue to determine whether the modeling was successful.After the model building was completed,rats in normal group became normal control group and the remaining rats in model group were randomly divided into model control group,low-dose group,middle-dose group,high-dose group and silybin group.The normal control group and model control group rats were given normal saline,and drug group rats were given corresponding doses of traditional Chinese medicine or western medicine by gastric administration.After 8 weeks,we measured rats weight,calculated the liver index and Lee’s index,observed hepatocytes HE staining and oil red O staining,tested serum ALT,AST,TG,TC and liver tissue TC,TG,FFA,MDA,SOD,GSH,TNFα,IL-6 levels,detected the expression of LXRa,SREBP-1c,FAS,DGAT2 by RT-PCR and Western Blot.Result:Clinical research: Comparison within the group: serum ALT,AST,GGT,ALP,TG,TC,HDL-C,LDL-C,FFA levels and TCM syndrome points,liver ultrasound image,fat attenuation CAP value,clinical comprehensive efficacy were all significantly improved after treatment compared with before treatment(P?0.05,P?0.01).There was no adverse reactions.Comparison between groups: after treatment,the treatment group was better than the control group in improving serum ALT,AST,TG,TC,LDL-C,FFA levels,TCM syndrome scores,liver ultrasound image,CAP value(P?0.05,P?0.01);The treatment group and the control group were equivalent in improving the serum GGT,ALP and HDL-C levels(P>0.05).The total effective rate was 85.71% in treatment group and 62.86%in control group,treatment group showed better therapeutic effect than control group(P?0.01).Experimental Study: NAFLD model rats were successfully established after12 weeks of HFD induction.In terms of body weight,liver index,Lee’s index and serum ALT,AST,TC,TG,compared with normal control group,the above indicators in model control group were significantly abnormal(P?0.01);Compared with model control group,the above indicators in each drug group were improved to different degrees(P?0.05,P?0.01);Compared with silybin group,the above indicators in high-dose group were significantly improved(P?0.05);Comparison within each dose group of TCM: the above indicators in high-dose groups were significantly improved than those in low-dose group(P?0.05,P?0.01),the indicators of body weight,Lee’s index,ALT,AST,TC,TG in middle-dose group were significantly improved than those in low-dose group(P?0.05)and the above indicators in high-dose group were significantly improved than those in middle-dose group(P?0.05).In terms of liver tissue TC,TG,FFA,MDA,SOD,GSH,TNFα and IL-6,compared with normal control group,the above indicators in model control group were significantly abnormal(P?0.01);Compared with model control group,the above indicators in each drug group were improved to different degrees(P?0.05,P?0.01);Compared with silybin group,the above indicators in high-dose group were significantly improved(P?0.05)and the FFA levels in middle-dose group were significantly reduced(P?0.05);Comparison within each dose group of TCM: the above indicators in high-dose group were significantly improved than those in low-dose group(P?0.05,P?0.01),the indicators of TC、TG、FFA、MDA、SOD、TNFα、IL-6 in middle-dose group were significantly improved than those in low-dose group(P?0.05)and the indicators of TC,FFA,MDA,SOD,GSH,TNFα,IL-6 in high-dose group were significantly improved than those in middle-dose group(P?0.05).In terms of liver tissue LXRα,SREBP-1c,FAS,DGAT2 m RNA and protein expression,compared with normal control group,the above indicators in model control group were significantly abnormal(P?0.01);Compared with model control group,the above indicators in each drug group were improved to different degrees(P?0.05,P?0.01);Compared with silybin group,the above indicators except for SREBP-1c in high-dose group were significantly reduced(P?0.05);Comparison within each dose group of TCM: the above indicators in middle-dose and high-dose group were significantly reduced than those in low-dose group(P?0.05,P?0.01),the above indicators except for SREBP-1c in high-dose group were significantly reduced than those in middle-dose group(P?0.05).Conclusion:1.Liver dysfunction with turbidity endogenesis are the key factors and central link of NAFLD.The principle of soothing liver and removing turbidity matches the basic mechanism of NAFLD,which can be used for treatment.2.Yinqi Tiaozhi Decoction can alleviate NAFLD condition,relieve symptoms and signs,restore liver function,reduce blood lipids,improve liver ultrasonography and CAP value.It is effective,safe and reliable.3.Yinqi Tiaozhi Decoction played a good therapeutic role on NAFLD rats induced by HFD.The mechanism of action is related to the regulation of LXRαpathway,and then changes the downstream targets of SREBP-1c,FAS,DGAT2,reduces liver lipid accumulation,alleviates liver tissue oxidative stress levels and inflammatory responses,decreases serum transaminase and blood lipid levels,and improves liver histopathology.The effect of high-dose group of traditional Chinese medicine was the most significant,suggesting that a higher concentration of Yinqi Tiaozhi Decoction could better regulate the LXRα pathway and relieve the disease.