摘要
[目的]探讨麝香保心丸(SBP)对糖尿病视网膜病变(DR)大鼠线粒体自噬及PTEN诱导激酶蛋白1(PINK1)/细胞质E3-泛素连接酶(Parkin)信号通路的影响。[方法]建立DR大鼠模型,将大鼠随机分为正常组(Control组)、模型组(DR组)、麝香保心丸低剂量组(SBP-L组)、麝香保心丸高剂量组(SBP-H组)、麝香保心丸高剂量+雷帕霉素组(SBP-H+RAP组)。检测各组大鼠空腹血糖(FPG)及总胆固醇(TC)、三酰甘油(TG)、低密度脂蛋白胆固醇(LDL-C)水平;苏木精-伊红(HE)染色观察各组大鼠DR情况;酶联免疫吸附实验(ELISA)检测各组大鼠血管内皮生长因子(VEGF)、高迁移率族蛋白1(HMGB1)水平;透射电镜观察各组大鼠细胞线粒体形态;免疫组化检测线粒体自噬相关蛋白选择性自噬接头蛋白sequestosome-1(SQSTM1/P62)、微管相关蛋白轻链3Ⅱ(LC3-Ⅱ)的表达;蛋白免疫印迹法检测各组大鼠PINK1、Parkin蛋白表达。[结果]与Control组比较,DR组视网膜结构破坏严重,细胞排列紊乱,水肿明显,细胞间隙变宽,线粒体损伤、肿胀明显,形态异常,FPG、TC、TG、LDL-C、VEGF、HMGB1、LC3-Ⅱ、PINK1、Parkin表达水平显著升高,SQSTM1/P62表达水平显著降低(P<0.05);与DR组比较,SBP-L组、SBP-H组视网膜细胞结构改善,细胞排列逐渐整齐,细胞水肿减轻,细胞、线粒体形态均趋于正常,线粒体肿胀减轻,FPG、TC、TG、LDL-C、VEGF、HMGB1、LC3-Ⅱ、PINK1、Parkin表达水平显著降低,SQSTM1/P62表达水平显著升高(P<0.05);与SBP-H组相比,SBP-H+RAP组视网膜细胞结构破坏加重,细胞排列紊乱,水肿明显,线粒体肿胀加重,形态异常,FPG、TC、TG、LDL-C、VEGF、HMGB1、LC3-Ⅱ、PINK1、Parkin表达水平显著升高,SQSTM1/P62表达水平显著降低(P<0.05)。[结论]麝香保心丸通过抑制PINK1/Parkin信号通路抑制DR大鼠线粒体自噬。
[Objective]To investigate the effect of Shexiang Baoxin Pill(SBP)on mitochondrial autophagy and PTEN-induced putative kinase 1(PINK1)/cytoplasmic E3 ubiquitin-ligase enzyme(Parkin)signaling pathway in diabetic retinopathy(DR)rats.[Methods]DR rats were separated into normal group(control group),model group(DR group),low dose Shexiang Baoxin Pill group(SBP-L group),high dose Shexiang Baoxin Pill group(SBP-H group),and high dose Shexiang Baoxin Pill+rapamycin group(SBP-H+RAP group).The levels of fasting blood sugar(FPG),total cholesterol(TC),triglyceride(TG)and low-density lipoprotein cholesterol(LDL-C)of blood lipids were detected in rats of each group;HE staining was applied to observe diabetic retinopathy of rats in each group;transmission electron microscopy was applied to observe the mitochondrial morphology of rat cells in each group;immunohistochemistry was applied to detect the expression of mitochondrial autophagy related proteins SQSTM1/P62,LC3-Ⅱ;Western blot was applied to detect the expression of PINK1 and Parkin proteins of rats in each group.[Results]Compared with the control group,the retinal structure of the DR group was severely damaged,with disordered cell arrangement,obvious edema,widened cell gaps,obvious mitochondrial damage and swelling,and abnormal morphology,the expression levels of FPG,TC,TG,LDL-C,LC3-Ⅱ,PINK1,and Parkin were obviously increased,the expression level of SQSTM1/P62 was obviously reduced(P<0.05);compared with the DR group,the structure of retinal cells in the SBP-L and SBP-H groups improved,the cell arrangement gradually became neat,the cell edema decreased,the morphology of cells and mitochondria tended to be normal,and mitochondrial swelling decreased,the expression levels of FPG,TC,TG,LDL-C,LC3-Ⅱ,PINK1,and Parkin were obviously reduced,the expression level of SQSTM1/P62 was obviously increased(P<0.05);compared with the SBP-H group,the structural damage of retinal cells in SBP-H+RAP group worsened,with disordered cell arrangement,obvious edema,increased mitochondrial swelling,and abnormal morphology,the expression levels of FPG,TC,TG,LDL-C,LC3-Ⅱ,PINK1,and Parkin were obviously increased,the expression level of SQSTM1/P62 was obviously reduced(P<0.05).[Conclusion]Shexiang Baoxin Pill can inhibit mitochondrial autophagy in diabetic retinopathy rats by inhibiting PINK1/Parkin signaling pathway.
作者
余玲
杨洋
段程慧
董瑞鸿
桑艳红
YU Ling;YANG Yang;DUAN Chenghui;DONG Ruihong;SANG Yanhong(Department of Endocrinology,the Fifth Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)
出处
《天津中医药》
CAS
2024年第3期366-371,共6页
Tianjin Journal of Traditional Chinese Medicine
基金
和黄科研基金-心血管专项(2018-2019)资助项目
