| With the increasing improvement of modern quality of life and changes in dietary structure,abnormal glucose and lipid metabolism occurs frequently in the body,thus increasing the prevalence of diabetes and hyperlipidemia year by year.Insulin resistance is one of the main pathogenesis of type 2 diabetes,and it can also cause glucose and lipid metabolism disorders.At this stage,there are many drugs that regulate abnormal glucose and lipid metabolism,but there are adverse reactions such as hypoglycemia,weight gain and gastrointestinal reactions,Therefore,people are eager to seek a kind of hypoglycemic drugs that are safe,effective and improve glucose and lipid metabolism disorders.This study mainly explored the effect of robustic on the glucose and lipid metabolism of insulin resistant HepG2 cells from in vitro experiments,evaluated the hypoglycemic effect and hypolipidemic effect of robustic on insulin resistant HepG2 cells,and discussed its mechanism of action.It lays the foundation for the development of new drugs for regulating abnormal glucose and lipid metabolism.In this study,the PNPG method was used to evaluate the inhibitory effect of robustic on α-glucosidase activity.Studies preliminarily found that robustic had inhibitory activity on α-glucosidase,and synergistically inhibited α-glucosidase activity with acarbose.The proliferation of robustic on HepG2 cells was investigated by MTT method,and provided a reference for the drug dosage of robustic in subsequent experiments.The experimental results showed that 100 μmol/L,50 μmol/L,25μmol/L,12.5 μmol/L,and 6.25 μmol/L of robustic had no growth inhibitory effect on HepG2 cells.Therefore,100 μmol/LL,50 μmol/L,25 μmol/L of robustic were selected for follow-up studies.Oleic acid was selected as the inducer to induce HepG2 cell model,and the optimal oleic acid induction concentration and time were selected by detecting cell proliferation,oil red O staining,observing and calculating lipid accumulation rate,the optimal oleic acid induction concentration was 0.5 mmol/L and the induction time was 24 h,the establishment of the insulin resistance cell models was investigated by glucose consumption experiments.Intervention of 100 μmol/L,50 μmol/L and 25 μmol/L of robustic in insulin resistance HepG2 cell model,to detect that robustic can improve lipid accumulation in insulin-resistant cells,improve lipid metabolism and liver function damage effect.The study showed that robustic in each dose group could improve the lipid accumulation caused by oleic acid,reduce the number of lipid droplets in HepG2 cells,and make the color of lipid droplets lighter.Compared with the model group,each dose groups of robustic could significantly reduce the contents of TC and TG in insulin-resistant cells(P<0.01),and could significantly reduce the ALT and AST contents of insulin-resistant cells(P<0.01).The high-dose and middle-dose groups of robustic could significantly reduce the LDH content of insulin-resistant cells(P<0.01),it has the effect of lowering blood lipid and improving liver function damage.Through glucose consumption experiment,glycogen content detection and2-NBDG fluorescence staining experiment,the hypoglycemic activity of robustic and its effect on insulin resistance were investigated.The results of the study showed that robustic can significantly(P<0.01 or P<0.05)promote the glucose consumption of insulin-resistant cells,increase the glycogen content of insulin-resistant cells,increase the insulin sensitivity index and glucose uptake.It has the effect of lowering blood sugar and improving insulin resistance.The m RNA and protein expressions of IRS-1,PI3 K p85,AKT and GLUT4 in insulin-resistant cells were detected by reverse transcription real-time fluorescent quantitative PCR(RT-qPCR)and Western blotting(Western blot).The results showed that robustic could up-regulate the m RNA expressions of IRS-1,PI3 K p85,AKT and GLUT4 in insulin-resistant cells(P<0.01 or P<0.05),as well as up-regulate the protein expression of IRS-1,PI3 K p85 and AKT(P<0.01).Therefore,it is inferred that the mechanism of action of drubic acid in improving abnormal glucose and lipid metabolism may be that it activates the IRS-1/PI3 K p85/AKT/GLUT4 signaling pathway by regulating the enzymatic activity of fatty acid transport-related proteins,thereby achieving hypoglycemic and hypolipidemic effects.The above results show that robustic has inhibiting α-glucosidase inhibitory activity,reduces the blood lipid level of insulin resistance model cells,protects liver tissue damage,promotes glucose consumption,increases insulin sensitivity index and protect liver tissue damage,thereby reduces blood lipid,blood sugar and improves glucose and lipid metabolism disorders.The mechanism of the above-mentioned effects may be that robustic can improve the excess fatty acids in cells,increase the enzymatic activity of fatty acid transport-related proteins,and promote the absorption and excretion of fatty acids in cells,thereby activating IRS-1 and making it interact with the PI3 K p85 subunit.Combined with each other,the downstream PI3 K is activated,and the signal is transduced to the AKT molecule,so that it can combine with GSK-3,accelerate the transport of GLUT4,improve the uptake and utilization of glucose,thereby improving insulin resistance and improving the effect of glucose and lipid metabolism.This study will provide experimental data and basis for the development of robustic as a drug for improving abnormal glucose and lipid metabolism. |
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