| Objective:(1)Clinical study:To evaluate the clinical efficacy and safety of Tangyiping in the treatment of patients with liver stagnation and spleen deficiency and phlegm and blood stasis with impaired glucose tolerance.(2)Experimental study:In vitro intervention of high glucose damaged or miR-204 overexpressed mouse pancreatic islet?cells by Tangyiping,observe the?cell viability,apoptosis and early apoptosis,glucose stimulated insulin secretion,miR-204 expression,TXNIP and MafA expression,confirm the mechanism of action of Tangyiping to improve islet?cell function.Methods:(1)Clinical study:70 patients with outpatients were randomly divided into two groups.The patients were divided into two groups:the sputum group and the Tangyiping group.The Tangyiping group was treated with Tangyiping,and the bismuth group was treated with metformin.Intervention to observe the effects of glucosinolate on fasting and postprandial glucose tolerance,glucose tolerance,insulin release,insulin sensitivity and?-cell function,glycosylated hemoglobin,blood lipids and TCM syndrome scores in patients with impotence and spleen deficiency The impact of various indicators and the evaluation of its safety.(2)Experimental study:The optimal concentration of melamine in the min6cells was screened by CCK-8 method,and the cells were divided into low glucose(LG),high glucose(HG),high glucose+Tangyiping(T),high.glucose+miR-204-5P mimic(MIM),high glucose+miR-204-5P mimic+Tangyiping(MIM+T),high glucose+miR-204-5P mimic negative control(MNC),CCK-8 kit was used to detect cell viability,JC-1 kit was used to detect mitochondrial membrane potential,Annexin V-FITC cell apoptosis kit was used to detect apoptosis,ELISA kit was used to detect glucose-stimulated insulin secretion,and RT-PCR was used to detect TXNIP.mRNA,MafA mRNA and miR-204expression,western-blot assay for TXNIP,MafA protein expression.Results:(1)Clinical study:A total of 34 patients in the Tangyiping group and 33 patients in the sputum group completed the study.(1)Blood glucose:Compared with before intervention,there was no significant difference in fasting blood glucose(P>0.05).There was no significant difference in fasting blood glucose between groups(P>0.05).Compared with before intervention,8 weeks and 12 weeks before intervention.The 2h PG was significantly decreased(P<0.05),indicating that both groups of interventions significantly reduced 2h PG by 8 weeks,and continued intervention could maintain glycemic control.(2)Insulin release:There was no significant difference in insulin between 0h,1h and 2h before treatment(P>0.05).Compared with before treatment,the area under FINS,1h insulin,2h insulin and insulin curve were significantly lower after treatment(P<0.05),after treatment,the area under 1h insulin,2h insulin and insulin curve in the Tangyipingline group was higher than that in the double sputum group(P<0.05),indicating that both groups could significantly reduce fasting insulin levels in IGT patients after treatment,and 1h.Insulin,2h insulin level and area under the insulin curve,and metformin was superior to Tangyiping in reducing the 1h,2h insulin level and the area under the insulin curve(P<0.05).(3)Glucose tolerance:PG,2h PG and AUCG were significantly decreased at 1h after treatment(P<0.05),and there was no significant difference in FPG.There was no significant difference between the two groups(P>0.05).It is indicated that Tangyiping can effectively reduce the area under the postprandial blood glucose and blood glucose curve of patients with IGT.(4)Islet?-cell function and insulin resistance:There was no significant difference between the two groups before treatment(P>0.05).Compared with before treatment,the?I60/?G60,HOMA-IR,MCBI,AUCG/I,MCBI were significantly decreased(P<0.05)and IAI was significantly increased(P<0.05),indicating that metformin can improve insulin.The role of resistance increased the insulin action index.After treatment with glucosinolate,?I60/?G60,HOMA-IR and AUCG/I were significantly decreased(P<0.05),IAI was significantly increased(P<0.05),and MCBI was slightly elevated,but The difference was not statistically significant(P>0.05).Compared with the treatment group,the?I60/?G60 and AUCI/G were increased after treatment in the Tangyiping group(P<0.05),indicating that glucosinolate was lower in the early secretory function and insulin sensitivity of islet?cells than metformin,MCBI.Elevated,indicating that the total insulin secretion of OGTT in the Tangyipingzide group was higher than that in the biguanide group.(5)Glycated hemoglobin:Both groups of interventions can reduce glycated hemoglobin(P<0.05),and the effect of reducing glycosylated hemoglobin in the two groups was comparable,no significant difference(P>0.05).(6)Blood lipids:There were no significant differences(P>0.05)between the two groups before the intervention,and they were comparable.Compared with before intervention,TC and TG were significantly decreased(P<0.05),indicating that Tangyiping and bismuth intervention group could improve TC and TG,and there was no difference between them(P>0.05).(7)Progress of the disease:After the intervention,the disease progress of the two groups was counted.Most of the patients were reversed to NGT,some of them maintained IGT,and a few of them progressed to DM.There was no significant difference between the two groups(P>0.05).It is suggested that metformin and Tangyiping have an average effect of reversing IGT,and the effects are similar.(8)improve blood glucose efficacy:There was no significant difference in the effective rate of blood glucose between the two groups(P>0.05),suggesting that the two groups are close to the effect of lowering blood glucose.(9)TCM syndrome scores:There was no significant difference in the total scores of the two groups before the intervention(P>0.05).Compared with the pre-intervention,the syndrome scores of the two groups were significantly lower(P<0.05).Compared with the stagnation group,the syndrome scores were significantly lower(P<0.05),indicating that both groups can improve the syndrome scores of patients.Tangyiping is superior to metformin in improving symptoms.(10)TCM Syndrome Points:Both groups can improve TCM syndrome scores,and the effect of Glucose Treatment on TCM Syndrome is significantly better than that of Shuangqi Group(P<0.05).?11 TCM syndrome single points:Both groups can improve the individual syndrome scores of TCM,and the effect of Tangyipingogenesis on TCM syndromes is significantly better than that of Shuangqi group(P<0.05).?12 Safety:No serious adverse reactions were reported in either group.(2)Experimental study:(1)The optimal intervention concentration of Tangyiping:the optimal concentration of Tangyiping was 5×10-4mg/m L.(2)Cell viability comparison:The cell viability of HG group was significantly better than that of LG group after drug intervention,suggesting that high glucose damages the activity of min6 cells.Compared with HG group,the cell viability of T group is significantly increased,MIM group is significantly decreased.Compared with MIM group,the cell viability of MIM+T group was significantly increased.(3)Mitochondrial membrane potential:Compared with LG group,mitochondrial membrane potential was significantly decreased in HG group(P<0.05).Compared with HG group,mitochondrial membrane potential was significantly increased in T group(P<0.05),The mitochondrial membrane potential of MIM mitochondria was decreased(P<0.05).Compared with MIM group,the mitochondrial membrane potential of MIM+T group was significantly decreased(P<0.05).(4)Apoptosis:Compared with LG group,the apoptosis rate of HG group was significantly increased.Compared with HG group,the apoptosis rate of T group was significantly decreased,and the apoptosis rate of MIM was significantly increased.Overexpression of miR-204-5P promoted apoptosis of min6.Compared with MIM group,the apoptosis rate of MIM+T group was significantly lower(P<0.05).(5)Glucose-stimulated insulin secretion:compared with LG group,HG group basal insulin and GSIS insulin were significantly lower(P<0.05),compared with HG group,T group basal insulin secretion and GSIS insulin were significant elevated(P<0.05),MIM basal insulin and GSIS were significantly lower(P<0.05).Compared with MIM group,MIM+T group and GSIS insulin were significantly increased(P<0.05).(6)TXNIP MafA mRNA expression:Compared with LG group,TXNIP mRNA expression was significantly increased in HG group(P<0.05),MafA mRNA expression was significantly decreased(P<0.05);compared with HG group,T group TXNIP mRNA expression was significantly decreased(P<0.05),MafA mRNA expression was significantly increased(P<0.05);TXNIP mRNA was significantly increased in MIM group(P<0.05),MafA mRNA expression was significantly decreased(P<0.05);Compared with the M group,the expression of TXNIP mRNA in MIM+T group was significantly decreased(P<0.05),and the expression of MafA was significantly increased(P<0.05).(7)miR-204-5P expression:Compared with LG group,the expression of miR-204-5P in HG group was significantly increased(P<0.05).Compared with HG group,the expression of miR-204-5P in T group was significant decreased.The expression of miR-204-5P in MIM group was significantly increased(P<0.05).Compared with MIM group,the expression of miR-204-5P was significantly decreased in MIM+T group(P<0.05).(8)TXNIP,MafA protein expression:Compared with LG group,TXNIP expression in HG group was significantly increased(P<0.05),MafA protein expression was significantly decreased(P<0.05),suggesting that high glucose can increase TXNIP protein expression and inhibit MafA protein expression;Compared with HG group,TXNIP protein expression was significantly decreased in T group(P<0.05),MafA protein expression was significantly increased(P<0.05);TXNIP protein was significantly increased in MIM group(P<0.05).The expression of MafA protein was significantly decreased(P<0.05).Compared with MIM group,the expression of TXNIP protein in MIM+T group was significantly decreased(P<0.05),and the expression of MafA was significantly increased(P<0.05).Conclusions:(1)Clinical study:Tangyiping can improve postprandial blood glucose,regulate insulin secretion,improve insulin resistance,reduce glycated hemoglobin,lower total cholesterol and triglycerides,improve patient syndrome scores,and improve safety.(2)Experimental study:Tangyiping can improve the activity of islet?cells with high glucose damage,improve apoptosis and early apoptosis,improve insulin secretion,decrease the expression of miR-204,TXNIP and increase the expression of MafA,and protect the function of islet?cells.It might be achieved by adjusting the TXNIP/miR-204/MafA pathway. |
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