Objective To investigate the ameliorative effect of Qihuakeli,a Hani formula,on glycolipid metabolism in type 2 diabetes mellitus by in vivo and in vitro experiments.Methods Rat liver mesenchymal stromal cells(BRL-3A)were inoculated in six-well plates and divided into blank,palmitic acid,fenofibrate,and Qihuakeli serum-containing 5.4,10.8,and 21.6 g/kg groups.Except for the blank group,the remaining groups were intervened with 0.2 mmol/L palmitic acid(PA)for 24 hour,and then added with drug-containing serum,and then continued to incubate for 24 hour.The proliferation rate of BRL-3A cells in each group was determined.Total cholesterol(T-CHO),low-density lipoprotein cholesterol(LDL-C),and high-density lipoprotein cholesterol(HDL-C)concentrations in the supernatant of each cell group were measured,cell culture medium was aspirated and discarded,triglyceride(TG)concentration in the cell lysate.The lipid content of the cells was determined by measuring and staining with red oil.Meanwhile,45 rats were taken and divided into blank group,model group,fenofibrate group(0.225 g/kg),Qihuakeli compound 5.4 g/kg group,and Qihuakeli compound 10.8 g/kg group,the blank group was given normal feed and the rest of the groups were given high-fat feed for 42 day.Beginning on the 43rd day,each group,except the blank group,was injected with a single intraperitoneal injection of Starting from the 43rd day,except the blank group,each group was given a one-time intraperitoneal injection of 0.25%streptozotocin(STZ)solution,and at the same time,the corresponding drugs were given by gavage for 14 day.The rats'weight gain and liver index were measured.Serum fasting blood glucose(FBG)and fasting insulin(FINS)were detected,and the insulin resistance index(ISI)was calculated.Serum free fatty acid(FFA)levels and tumor necrosis factor-α(TNF-α)in liver tissue were also detected.HE staining was used to detect pathological changes in the pancreas.Pathological changes were observed in the tissues,and islet α and β cell expression was detected by immunohistochemistry.Results Compared to the PA group,the accumulation rate of BRL-3A cells was significantly higher(P<0.01)in the 10.8 and 21.6 g/kg Qihuakeli-containing serum groups.The levels of T-CHO,LDL-C and TG in the 5.4 and 21.6 g/kg serum groups were significantly lower(P<0.05),and HDL-C levels significantly increased(P<0.05).Oil red staining results showed that lipids in the cytoplasm of the 5.4,10.8 and 21.6 g/kg.Qihuacel-containing groups significantly reduced.Compared to the model group,the body weight of the 10.8 g/kg group containing Qihuakeli granules increased significantly(P<0.05).The liver index of the 5.4 g/kg group containing Qihuakeli decreased significantly(P<0.05).The serum indices of FBG,FINS,FFA and insulin resistance of the 5 g/kg group containing Qihuakeli decreased significantly(P<0.05).In the 5.4,10.8 g/kg groups,all serum FBG,FINS,FFA and insulin resistance indices significantly reduced in the 5.4 and 10.8 g/kg Qihuakeli groups(P<0.05 or P<0.01).TNF-α levels were significantly reduced(P<0.01).HE staining showed that a small number of lymphocytes were scattered in the pancreatic ducts and perivascular area of the rats in the Qihuakeli 5.4 and 10.8 g/kg groups,the local vasodilatation was observed,the number of pancreatic islet cells and the area of islet cells significantly increased.Immunohistochemical study was further used.The results of immunohistochemistry showed that the area of pancreatic islet α-cells significantly reduced and the area of pancreatic islet β-cells significantly increased in Qihuakeli 5.4 and 10.8 g/kg groups.Conclusion Qihuakeli compound improved glucose-lipid metabolism in T2DM,probably by improving the function of pancreatic islet cells,increasing the sensitivity of insulin to blood glucose,improving insulin resistance,decreasing the secretion of insulin and glucagon,and thus lowering the level of fasting blood glucose.Meanwhile,by decreasing the content of TNF-α,inhibiting lipolysis in the body,and promoting the uptake of FFA by adipocytes,and further lowering the FFA.Thus,it regulates the levels of TG,T-CHO,HDL-C and LDL-C,improves the abnormalities of glucose and lipid metabolism,and alleviates T2DM.

Objective To investigate the ameliorative effect of Qihuakeli,a Hani formula,on glycolipid metabolism in type 2 diabetes mellitus by in vivo and in vitro experiments.Methods Rat liver mesenchymal stromal cells(BRL-3A)were inoculated in six-well plates and divided into blank,palmitic acid,fenofibrate,and Qihuakeli serum-containing 5.4,10.8,and 21.6 g/kg groups.Except for the blank group,the remaining groups were intervened with 0.2 mmol/L palmitic acid(PA)for 24 hour,and then added with drug-containing serum,and then continued to incubate for 24 hour.The proliferation rate of BRL-3A cells in each group was determined.Total cholesterol(T-CHO),low-density lipoprotein cholesterol(LDL-C),and high-density lipoprotein cholesterol(HDL-C)concentrations in the supernatant of each cell group were measured,cell culture medium was aspirated and discarded,triglyceride(TG)concentration in the cell lysate.The lipid content of the cells was determined by measuring and staining with red oil.Meanwhile,45 rats were taken and divided into blank group,model group,fenofibrate group(0.225 g/kg),Qihuakeli compound 5.4 g/kg group,and Qihuakeli compound 10.8 g/kg group,the blank group was given normal feed and the rest of the groups were given high-fat feed for 42 day.Beginning on the 43rd day,each group,except the blank group,was injected with a single intraperitoneal injection of Starting from the 43rd day,except the blank group,each group was given a one-time intraperitoneal injection of 0.25%streptozotocin(STZ)solution,and at the same time,the corresponding drugs were given by gavage for 14 day.The rats'weight gain and liver index were measured.Serum fasting blood glucose(FBG)and fasting insulin(FINS)were detected,and the insulin resistance index(ISI)was calculated.Serum free fatty acid(FFA)levels and tumor necrosis factor-α(TNF-α)in liver tissue were also detected.HE staining was used to detect pathological changes in the pancreas.Pathological changes were observed in the tissues,and islet α and β cell expression was detected by immunohistochemistry.Results Compared to the PA group,the accumulation rate of BRL-3A cells was significantly higher(P<0.01)in the 10.8 and 21.6 g/kg Qihuakeli-containing serum groups.The levels of T-CHO,LDL-C and TG in the 5.4 and 21.6 g/kg serum groups were significantly lower(P<0.05),and HDL-C levels significantly increased(P<0.05).Oil red staining results showed that lipids in the cytoplasm of the 5.4,10.8 and 21.6 g/kg.Qihuacel-containing groups significantly reduced.Compared to the model group,the body weight of the 10.8 g/kg group containing Qihuakeli granules increased significantly(P<0.05).The liver index of the 5.4 g/kg group containing Qihuakeli decreased significantly(P<0.05).The serum indices of FBG,FINS,FFA and insulin resistance of the 5 g/kg group containing Qihuakeli decreased significantly(P<0.05).In the 5.4,10.8 g/kg groups,all serum FBG,FINS,FFA and insulin resistance indices significantly reduced in the 5.4 and 10.8 g/kg Qihuakeli groups(P<0.05 or P<0.01).TNF-α levels were significantly reduced(P<0.01).HE staining showed that a small number of lymphocytes were scattered in the pancreatic ducts and perivascular area of the rats in the Qihuakeli 5.4 and 10.8 g/kg groups,the local vasodilatation was observed,the number of pancreatic islet cells and the area of islet cells significantly increased.Immunohistochemical study was further used.The results of immunohistochemistry showed that the area of pancreatic islet α-cells significantly reduced and the area of pancreatic islet β-cells significantly increased in Qihuakeli 5.4 and 10.8 g/kg groups.Conclusion Qihuakeli compound improved glucose-lipid metabolism in T2DM,probably by improving the function of pancreatic islet cells,increasing the sensitivity of insulin to blood glucose,improving insulin resistance,decreasing the secretion of insulin and glucagon,and thus lowering the level of fasting blood glucose.Meanwhile,by decreasing the content of TNF-α,inhibiting lipolysis in the body,and promoting the uptake of FFA by adipocytes,and further lowering the FFA.Thus,it regulates the levels of TG,T-CHO,HDL-C and LDL-C,improves the abnormalities of glucose and lipid metabolism,and alleviates T2DM.

Objective:To establish a method to determine the content of ursolic acid in Zhike Chuanbei Pipa syrups. Methods:An HPLC method was used with a Cosmosil-C18 column(250 mm × 4. 6 mm, 5 μm). The mobile phase was methanol-water-glacial acetic acid-triethylamine(90∶10∶0. 03∶0. 06) with a flow rate of 1. 0 ml·min-1 , the column temperature was 25℃, and the detec-tion wavelength was set at 215nm. Results:The linear range of ursolic acid was 16-80μg·ml-1(r=0. 999 5, n=5), and the aver-age recovery was 98. 67℅ (RSD=1. 22, n=6). Conclusion:The method is simple, accurate and reliable with good specificity and reproducibility, and it can be used in the quantitative determination of ursolic acid in Zhike Chuanbei Pipa syrups.