Study On Extraction Technology,Quality Standards,Regulation Of Blood Glucose And Mechanism Of Gesang Jiangtang Capsule

Objective:In this study,a series of studies were conducted on the extraction process,quality standards,regulation of blood glucose and its mechanisms of Gesang Jiangtang Capsules by applying modern Chinese medicinal preparations and traditional Chinese medicine pharmacology techniques,in order to provide an experimental basis for the development of a state-level auxiliary hypoglycemic health food.Method:First,about studing on extraction technology:The water extraction technology,of MORI FRUCTUS and so on,was optimized by orthogonal experiment with amount of water,extraction time and extraction times as factor,using the extraction volume of total crude polysaccharide and the yield of extract as index.The thanol extraction technology,of PUERARIAE LOBATAE RADIX and so on,was optimized by orthogonal experiment with amount of ethanol,extraction time and extraction times as factor,using the extraction volume of puerarin,total saponins and the yield of extract as index.Secondly,about studing on quality strander:UV spectrophotometric method was used for determining polysaccharides,total saponins and total flavonoids of GSC.10 batches of GSC were detected and recorded by UPLC.Thirdly,about studing on regulation of blood glucose:Firstly,using a normal mouse model,continuous gavage with 1000mg/kg BW GSC for 30 days to observe its effects on fasting blood glucose and body weight;Secondly,using a hyperglycemia mouse model induced by alloxan,continuous gavage with 1000,500,and 250 g/kg BW GSC for 30 days to observe its effects on fasting blood glucose,glucose tolerance,and body weight and so on;Thirdly,using type 2 diabetic rat model induced by a high-fat,high-sugar diet combined with low-dose streptozotocin,continuous gavage with 500,250,125 g/kg BW GSC for 38 days to observe its effects on fasting blood glucose,blood lipids,and body weight and so on;Fourthly,about studing on regulation of blood glucose mechanism:using type 2 diabetic rat model induced by a high-fat,high-sugar diet combined with low-dose streptozotocin,serum markers of oxidative stress,hepatic and muscle glycogen content,and the insulin signal transduction pathway PI3K/AKT key protein expression in liver were determined.Result:First,about studing on extraction technology:Optimal ethanol extraction technology was as follows:6 folds of ethanol,extracting for 2 times,1h each time.The water extraction technology was as follows:6 folds of water,extracting for 2 times,1h each time.Secondly,about studing on quality strander:Average polysaccharides content in three batches of GSC was 45.55 mg/g,the average recovery was 99.64%and the RSD was less than 5%.Average total saponins ginsenoside Re content was 65.41 mg/g,the average recovery was 96.97%and the RSD was less than 5%.Average total flavonoids content was 2.61 mg/g,the average recovery was 96.70%and the RSD was less than 5%.UPLC fingerprints of 10 batches of GSC were established and 15 common peaks were confirmed.Based on the retention time of reference substances,7 constituents were identified.Thirdly,about studing on regulation of blood glucose:About the normal mouse model,GSC had no significant(P>0.05)effect on the blood glucose and body weight of normal mouse.About the alloxan-induced diabetic mice,the fasting blood glucose and glucose tolerance of the mice in GSC500 and GSC250 were significantly(P<0.05)different as compared with the diabetic control group,but the weight of mice had no significant(P>0.05)effect.About type 2 diabetic rat model,compared with the model group,fasting blood glucose,serum TC,TG,LDL-c,FFA levels in GSC500,GSC250 and MET200 rats were significantly reduced(P<0.05),serum HDL-c and insulin levels were significantly increased(P<0.05),but body weight had no significant effect(P>0.05).Fourthly,about studing on regulation mechanism:Compared with the model group,serum SOD,CAT and GSH-PX levels,hepatic glycogen and muscle glycogen content were significantly increased in GSC500,GSC250 and MET200 rats,but MDA levels were significantly lower than the model group(P<0.05 or P<0.01);Compared with the model group,the pathological changes of the pancreas and liver tissues of the rats in each administration group were reduced to varying degrees;Compared with the model group,The protein levels of PI3K,AKT,and GLUT-4 of livers of in GSC500,GSC250 and MET200 rats were significantly increased,but the expression of GSK-3βprotein was significantly decreased(P<0.05 or P<0.01).Conclusion:First,the optimal extraction process for this experiment is reasonable and feasible,which is used for the extraction of Chinese medicinal materials of GSC.The established colorimetric determination and UPLC fingerprint are scientific and reasonable,which were used for the quality control of GSC.Second,this experiment found that GSC has no significant effect on blood glucose in normal mice,but it can reduce fasting blood glucose and improve glucose tolerance in alloxan-included hyperglycemia mouse,and it can reduce fasting blood glucose in type 2 diabetic rats.Third,this experiment found that the mechanism of GSC in reducing fasting blood glucose of type 2 diabetic rats may be protecting islets,adjusting lipid metabolism of liver tissue,regulating oxidation/antioxidation balance,increasing glycogen synthesis,and regulating PI3K/AKT signal path.