The Protective Effect Of Tangnaikang On The Damage Of Pancreatic Islet Cell Line INS-1 Induced By Glycolipid Toxicity

OBJECTIVE:This experiment by in vitro culture islet beta cells glycolipids INS-1 poison model,using"qi and heat resistance of sugar"kang group intervention,to observe the sugar tolerance of glycolipids,under dynamic islet beta cells,apoptosis and insulin secretion,and further discuss from islet beta cells insulin signal transduction pathways downstream FOXO1 sugar resistant kang of PI3K/Akt,the influence of such factors to explore the specific targets.It provides a theoretical basis for the treatment of early type 2 diabetes induced by glycolipidosis with traditional Chinese medicine.METHOD:1.Effect of the serum containing Tangnaikang on the activity of ins-1 cells damaged by glycolipidosisIn vitro,insulinoma cell line insulin-1 cells in rats were selected as the subjects of this study.Insulin-1 cells were cultured with 33.3mM glucose to establish a model of glucose toxicity,insulin-1 cells were cultured with 0.5mM palmitate to establish a model of lipid toxicity,and insulin-1 cells were cultured with 33.3mM glucose and 0.5mM palmitate to establish a model of glucose toxicity.The serum containing the drug was prepared,and the intervention was carried out at low(5%),medium(8%)and high(10%)doses of the drug.Metformin hydrochloride was used as the western medicine control,and high sugar,palmitic acid and high sugar combined palmitic acid were used as the model control.CCK-8 was used to detect the activity of ins-1 cells,flow cell fluorescence staining,and caspase-3 activity in each group.Apoptosis was observed,and insulin secretion function of ins-1 cells under glucose stimulation was measured by ELISA.2.The effect of serum containing Tangnaikang on insulin signal transduction pathway of ins-1 cells damaged by glycolipidosisThe serum containing the drug was prepared,and the intervention was carried out at low(5%),medium(8%)and high(10%)doses of the drug.Metformin hydrochloride was used as the western medicine control,and high sugar,palmitic acid and high sugar combined palmitic acid were used as the model control.Immunoblotting was used to determine the protein content of insulin receptor substrate-2(IRS2)and downstream PI3K related proteins:forkhead transcription factor(FOXOl),protein tyrosine phosphatase 1B(PTP1B)in the insulin signaling pathway,as well as the phosphorylation levels of IRS2(ser731),FOXO1(ser256)and P70s6k(htr389).RESULTS:1.Effect of the serum containing Tangnaikang on the activity of ins-1 cells damaged by glycolipidosis.High glucose,high fat and high sugar combined with high fat can significantly reduce the activity of ins-l1cells.After the intervention of Tangnaikang,compared with the model group,the cell viability,caspase-3 activity and BIS and GSIS values in the high-dose Tangnaikang group increased significantly.Low dose and medium dose tanecan improved the above indexes to varying degrees.Compared with the metformin control group,there was no significant difference between the two groups.2.Effect of the serum containing Tangnaikang on the insulin signal transduction pathway of ins-1 cells damaged by glycolipidosisIRS2 protein expression was decreased and phosphorylation level was increased in the group with high glucose,high fat and high sugar combined with high fat The phosphorylation of FOXO1(ser256)was decreased and the expression of PTP1B was increased.After the intervention of Tangnaikang,compared with the model group,the expression of IRS2protein was increased and the phosphorylation level was decreased in the high-dose Tangnaikang group.The phosphorylation levels of FOXO1(ser256)and P70s6k(htr389)were increased,and PTP1B protein expression was decreased.Compared with the metformin control group,there was no significant difference between the two groups.CONCLUSION:1.Tangnaikang can improve the activity of ins-1 cells stimulated by glycolipidosis,inhibit apoptosis,and improve the function of insulin secretion.2.Tangnaikang can increase the expression of IRS2 protein in the insulin signal transduction pathway of ins-1 cells stimulated by glycolipidosis,reduce the phosphorylation level of its silk/threonine,inhibit the expression of PTP1B protein,activate the PI3K/AKT signaling pathway,and activate the downstream FOXO1 factor,thereby improving the glucose and lipid metabolism.