Based On Mitophagy Discuss Effect Of Buqing Granules Medicated Sera Against The Human Lens Epithelial Cells Induced By High Glucose Damage Mitophagy

Objective:To observe the Buqing Granules regulation "mitochondrial autophagy" related signaling pathway to improve SRA01/04 cell damage induced by high-glucose and its molecular mechanism,to Buqing Granules treatment provide experimental basis for the clinical application of diabetic cataract.Methods:1.The serum containing Buqing Granules was prepared by adoping the method of serum pharmacology of TCM.Rats were divided into the blank group(distilled water,10 m L·kg-1),Qiju Dihuangwan(QJ)group,BQ group(4,2,1g·m L-1).there were 15 rats in each group.Bid intragastric administration,and at the end of the 7th day,blood sampling after intragastric administration 1h.Under the condition of high speed centrifugal preparation of medicated serum in 3000rpm·min-1.2.The immortalized human lens epithelial cells(SRA01/04)training preparation high-glucose model.SRA01/04 cells were divided into 6 group,Continue our preliminary research result,The tendency for 30 mmol·L-1 glucose as the optimum concentration of cell proliferation,used to simulate high-glucose environment intervention in the experimental group,respectively.10%blank serum group,high-glucose group+10%blank serum group,high-glucose+10%serum containing BQ 4,2,1 g·mL-1group,high-glucose+10%serum containing QJgroup.With CCK 8 method to detect different medicated serum of high-glucose induced SRA01/04 24 h cell proliferation activity of influence.3.Use medicated serum of the experimental group intervention model.Immunofluorescence techniques were used respectively to observe the expression of each cell mitochondria autophagy related proteins Parkin,Pink-1,P62.Using Western bolt testing each cell mitochondria autophagy related proteins,Parkin,P62,Pink-1,Bnip3,Nix expression changes.Using the Real-time PCR method to detect LC3-Ⅱ and LC3-Ⅰ mRNA expression.Result:1.CCK-8 shows high-glucose induced SRA01/04 cell proliferation activity,under the different concentration for Buqing Granules medicated serum of high-glucose induced SRA01/04 cell proliferation activity,and in the serum concentration of 2g·m L-1,the inhibition significantly(P<0.05).2.Immunofluorescence test results show that different SRA01/04 cell medicated serum group were visible expression of mitochondria autophagy related protein Parkin,P62,Pink-1,and more than the expression in the cytoplasm,different medicated serum groups of the protein expression differences.3.Western bolt test results showed that the Pink-1/Parkin signaling pathways in the study found that high-glucose induced SRA01/04 cells visible the expression of mitochondria autophagy related proteins Parkin,Pink-1,P62,give Buqing Granules medicated serum intervened the expression of Pink-1,Parkin were increased(P<0.01),the expression of P62 was reduce(P<0.01);Found in the study of Bnip3/Nix signal pathway,high-glucose induced SRA01/04 cells visible the expression of Bnip3,Nix and with the extension of injury time showed a trend of increase after decreases first,to 24 h after injury of peak and give Buqing Granules medicated serum intervened,the expression trend more significantly(P<0.01),and the effect of prolonged(P<0.01).4.Real-time PCR detection results show that different SRA01/04 cell medicated serum group were visible the expression of microtubule associated protein LC3-Ⅱ and LC3-Ⅰ mRNA,and LC3-Ⅱ and LC3-Ⅰ ratio showed a trend of increase after decreases first,and peaked during the 24 h after injury,to give Buqing Granules medicated serum intervened,the trend is more obvious(P<0.01),and the effect of prolonged(P<0.01).Conclusion:Buqing Granules can effectively restrain high-glucose induced SRA01/04 cell proliferation,reduce caused by SRA01/04 cell excessive proliferation of lens opacity,its mechanism may be related to intervention damage is related to the dynamic expression of mitochondrial autophagy related proteins.