The Study Of Effects And Mechanism Of Shugan Wendan Decoction In Improving Insulin Resistance In Metabolic Syndrome

Objective:To evaluate the effects of Shugan Wendan Decoction for improving insulin resistance in patients with metabolic syndrome(MS);To clear that Shugan Wendan Decoction could improve insulin resistance(IR),glucose and lipid metabolism,oxidative stress in MS rats through liver X receptora(LXRα)mediated autophagy/apoptosis.Method:1.Clinical trial:A randomized,controlled clinical trials was conducted.84 patients with MS of Qi stagnation and phlegm stasis syndrome were randomly divided into the control group and the treatment group.The control group received conventional Western medicine treatment,and the treatment group received conventional Western medicine combined with Shugan Wendan Decoction for 12 weeks.The weight,body mass index(BMI),blood pressure,blood lipids,fasting blood glucose(FPG),hemoglobin Alc(HbA1c),serum insulin(FINS),homeostasis model assessment of insulin resistance(HOMA-IR),superoxide dismutase(SOD),glutathione peroxidase(GSH-PX),malondialdehyde(MDA),TCM syndrome scores before and after treatment in the two groups were detected.And the efficacy of TCM syndromes in the two groups after treatment was evaluated.2.1 Animal Experiment 1:Intraperitoneal injection of streptozotocin(STZ)combined with high-fat,high-sugar and high-salt feed was used to replicate the MS rat model,and the SD rats were randomly divided into normal control group(NC group)and metabolic syndrome Model group(MS group),Shugan Wendan Decoction high-dose group(SGWD-H group),Shugan Wendan Decoction medium-dose group(SGWD-M group),Shugan Wendan Decoction low-dose group(SGWD-L group),metformin group(MF group).The SGWD-H group,SGWD-M group,and SGWD-L group were given with a dose of 23.58g/Kg/d,11.79g/Kg/d,5.90g/Kg/d Shugan Wendan Decoction each day by the oral administration,respectively.MF group was administrated with Metformin hydrochloride tablets(0.09g/Kg/d)by the oral administration,and the NC group and MS group were given an equal volume of normal saline by the oral administration.The drug intervention time was 4 weeks.After the intervention,the rats weight,body length and blood pressure were measured,HbAlc and FINS contents were detected by ELISA;FPG,TC,TG,LDL-C,HDL-C contents were detected by biochemical analyzer,and HOMA-IR and HE staining methods were calculated.The pathological changes of rat liver and vascular tissue were observed by hematoxylin-eosin staining.2.2 Animal Experiment 2:Experimental animals,grouping and drug intervention methods are the same as animal experiment 1.ELISA method was used to detect SOD,GSH-PX,MDA content,and western-blot method was used to detect rat liver tissue LXRα,LC3,Beclin-1,Bax,Caspase-3,Bcl-2 protein expression,QPCR method to detect the rat liver tissue LXRα,LC3,Beclin-1,Bax,Caspase-3,Bcl-2 mRNA expression.2.3 Animal Experiment 3:The NC group,MS group,SGWD-M group came from animal experiment 1.Moreover,Shugan Wendan Decoction+LXRa inhibitor group[SGWD+LXRα(-)group]was set up.The NC group and MS group were given an equal volume of normal saline by the oral administration,SGWD-M group was given Shugan Wendan Decoction(11.79g/Kg/d)by gavage,and the SGWD+LXRα(-)group was given Shugan Wendan Decoction(11.79g/Kg/d)by gavage+GSK2033[(30mg/kg/d),ip,qd]intraperitoneal injection,the intervention time is 4 weeks.The detection indicators and methods are the same as those of animal experiment one and two.Result:1.Clinical trials:① After the treatment,compared with the control group,the FPG,HbA1c,FINS,HOMA-IR,TC,TG,LDL-C,HOMA-IR,MDA levels in the treatment group reduced significantly(P<0.05),HDL-C,SOD,GSH-PX level increased significantly(P<0.05);②After the treatment,compared with the control group,the TCM syndrome score of the treatment group reduced significantly(P<0.05),and the efficacy of TCM syndrome increased significantly(P<0.05).2.1 Animal Experiment 1:①Compared with NC group,FPG,HbAlc,FINS,HOMA-IR,TC,TG,LDL-C,HOMA-IR,body mass,Lee’s index,SBP,DBP levels of rats in the MS group and SGWD-L group were significantly increased(P<0.05),HDL-C was significantly reduced(P<0.05),HE staining showed liver tissue steatosis and vacuolar degeneration,HE staining showed vascular tissue atherosclerosis;②Compared with the MS Group and SGWD-L group,FPG,HbAlc,FINS,HOMA-IR,TC,TG,LDL-C,HOMA-IR,body mass,Lee’s index level in the SGWD-H group,SGWD-M group,MF group were significantly increased(P<0.05),HDL-C was significantly reduced(P<0.05),HE staining showed that liver tissue steatosis and vacuolar degeneration reduced significantly,and HE staining showed that vascular tissue atherosclerosis reduced significantly.2.2 Animal Experiment 2:①Compared with the NC group,the serum SOD,GSH-PX and mRNA and protein expression of LXRα,LC3,Beclin-1,Bcl-2 in liver tissue in the MS and SGWD-L groups were significantly reduced(P<0.05),while serum MDA and and mRNA and protein expression of Bax,Caspase-3 in liver tissue were significantly increased(P<0.05);②Compared with MS group and SGWD-L group,SOD,GSH-PX and mRNA and protein expression of LXR α,LC3,Beclin-1,Bcl-2 in liver tissue in the SGWD-H group,SGWD-M group and MF groups were significantly increased(P<0.05),MDA and mRNA and protein expression of Bax,Caspase-3 in liver tissue were significantly reduced(P<0.05).2.3 Animal Experiment 3:Compared with the SGWD-M group,FPG,FINS,HOMA-IR,TC,TG,LDL-C,HOMA-IR,MDA and mRNA and protein expression of Bax,Caspase-3 in liver tissue in the SGWD+LXR α(-)group were significantly increased(P<0.05),HDL-C,SOD,GSH-PX and mRNA and protein expression LXR α,LC3,Beclin-1,Bcl-2 in liver tissue were significantly reduced(P<0.05).Conclusion:1.Clinical trials:Shugan Wendan Decoction could improve insulin resistance,glucose and lipid metabolism,Oxidative stress in patients with metabolic syndrome.2.Animal experiment:Shugan Wendan Decoction could improve insulin resistance,glucose and lipid metabolism,oxidative stress in rats with metabolic syndrome through LXR α mediated autophagy/apoptosis,and reduce liver steatosis and vascular atherosclerosis pathology change.