Pharmacodynamics Of Buqing Granules On Diabetic Cataract In Early And Middle Stage Of Db/db Mice And Study On Initiation Mechanism Of Delaying Damaged Mitochondria

Objective:To examine the protection effect of lens injury caused by early and middle diabetic cataract from Buqing granules endowed to db/db mice.To explore the signal transduction pathway?Pink1-Parkin signal pathway,Bnip3/Nix signal pathway and FUNDC1signal pathway?of Buqing granules through regulating the initiation stage of damaged mitochondria in"mitochondrial autophagy"to alleviate the molecular mechanism of lens damage resulted from diabetic cataract.To offer novel solutions for the nosogenesis and treatment of early and middle diabetic cataract.Methods:Choose db/db mice?6 weeks of age,being fed for 5 weeks?,give them treatment of Buqing granules and positive control drug—lycii and chrysanthemi and rehmanniae bolus,then observe their daily condition,glucose level and lipid level,and watch histopathological and pathological changes of their lens by light microscope.Apply the immunohistochemical method to detect the manifestation of PINK1,LC3 protein in the crystalline lens issues from db/db mice.Adopt the Western blot technology and the real-time quantitative PCR detecting system technology to test the manifestation of Bnip3,Nix and FUNDC1 in the crystalline lens issues of these mice.Use immortalized human lens epithelial cells?SRA01/04?and H2O2 for induced preparation model.Divide SRA01/04 cells into six groups,each group is intervened by drug-containing serum.Apply CCK-8 method to detect the effects of different drug-containing serum on the activity of SRA01/04 cells induced by hydrogen peroxide for.Respectively adopt the Western blot technology and the real-time quantitative PCR detecting system to detect the manifestation of Bnip3,Nix and FUNDC1 in human lens epithelial cells.Results:1.Pharmacodynamics research results of db/db mice at the early and middle stage of their diabetic cataract under the protection of Buqing granules.1.1 High Buqing granules concentration group improved the weight of db/db mice?two weeks after the treatment?.Buqing granules improved fasting blood sugar and lipids?P<0.05?.1.2 Result from the hematoxylin-eosin staining treatment showed:Buqing granules conducted obvious repairing and protective effects on DC lens epithelial cells at early and middle stage of db/db mice.1.3 Results of CCK-8 manifested that abnormal increase of activity of SRA01/04 cells induced by H₂O₂,the serum containing Buqing Granules at different concentrations protected the activity of SRA01/04 cells induced by H₂O₂,and the protective effect was significant when the high concentration of serum containing drugs was used to intervene?P<0.05?.2.Research results on the delayed mitochondrial initiation mechanism of lens tissue damage by Buqing granules2.1 Result of immunohistochemistry manifested that expressions of LC3 and Pink1,key proteins of mitochondrial autophagy could be spotted in DC lens of db/db mice at the early and middle stages.After giving Buqing granules,protein expressions of them were all increased?P<0.05?.2.2 Detection results of Western bolt showed that in the study of Pink-1/Parkin signaling pathway,the expression of mitochondrial autophagy-related proteins Bnip3,Nix and FUNDC1 was observed in DC lens of db/db mice in early and middle stages,and the expression of three proteins increased after the treatment with Buqing granules?P<0.05?.2.3 Results from real-time PCR showed that the expression of mitochondrial proteins Bnip3,Nix and FUNDC1 was observed in DC lens of db/db mice in the early and middle stages of the Pink-1/Parkin signaling pathway.The expression of the three proteins increased after the treatment of Buqing granules?P<0.05?.3.Research results on the mechanism of Buqing granules delaying damaged mitochondrial priming of lens epithelial cells3.1 Detection results of Western bolt manifested that the expression of mitochondrial autophagy-related proteins Bnip3,Nix and FUNDC1 could be observed in SRA01/04 cells induced by hydrogen peroxide,and the expression of three proteins increased after the intervention of Buqing granules containing serum?P<0.05?.3.2Real-time PCR results showed that the expression of mitochondrial autophagy-related proteins Bnip3,Nix and FUNDC1 could be observed in SRA01/04 cells induced by hydrogen peroxide.The expression of the three proteins increased after the intervention of Buqing granules containing serum?P<0.05?.Conclusion1.Decreased lens autophagy may cause diabetic cataract.2.Buqing Granules has protective effect on early and middle stage diabetic cataract in db/db mice;3.The curative effect of Buqing Granule on early and middle stage diabetic cataract is similar to that of Qiju Dihuang Pill.Among them,the high concentration of Buqing Granule is the best;4.Buqing Granules can promote the development of mitochondrial autophagy by up-regulating the expression of LC3,Pink1,Bnip3,Nix and FUNDC1 proteins,and activating the signal pathways of Pink1-Parkin,Bnip3/Nix and FUNDC1.