活血定眩胶囊含药血清对小鼠脑微血管内皮细胞bEnd.3自噬的影响  

Effects of serum containing Huoxue Dingxuan Capsule on bEnd.3 autophagy in mouse brain microvascular endothelial cells

作　　者：巩彦龙 宋敏 董万涛 董平 宋志靖 文皓楠 海云翔 GONG Yan-long;SONG Min;DONG Wan-tao;DONG Ping;SONG Zhi-jing;WEN Hao-nan;HAI Yun-xiang(traditional Chinese Medicine Clinical College,Gansu University of Chinese Medicine,Lanzhou 730000,China;Orthopedics Department,Affiliated Hospital of Gansu University of Chinese Medicine,Lanzhou 730000,China;College of Traditional Chinese Medicine,Inner Mongolia Medical University,Hohhot 010110,China)

机构地区：[1]甘肃中医药大学中医临床学院,兰州730000 [2]甘肃中医药大学附属医院骨科,兰州730002 [3]内蒙古医科大学中医学院,呼和浩特010110

出　　处：《中华中医药杂志》2020年第5期2653-2656,共4页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金地区科学基金项目(No.81760876);甘肃省高等学校科研项目(No.2017A-053)。

摘　　要：目的:探讨活血定眩胶囊含药血清对氧糖剥夺损伤(OGD)培养的bEnd.3细胞自噬的干预情况。方法:制备各大鼠含药血清,将bEnd.3细胞传代培养并建立OGD模型后,将其分为OGD模型组,活血定眩胶囊含药血清低、中、高剂量组及盐酸氟桂利嗪组,取bEnd.3细胞为对照组,进行相应的方法干预,干预后检测各组细胞丙二醛(MDA)浓度、乳酸脱氢酶(LD)活性,激光共聚焦扫描显微镜分析LC3点状聚集计数并计算灰度值,qRT-PCR方法检测自噬相关基因LC3Ⅱ、P62、Beclin-1 mRNA的表达。结果:与对照组比较,OGD模型组MDA浓度、LD活性、LC3Ⅱ表达量及LC3Ⅱ、P62、Beclin-1 mRNA表达均显著升高(P<0.05);与OGD模型组比较,经活血定眩胶囊含药血清中、高剂量干预后,各组MDA浓度、LD活性、LC3Ⅱ表达量及LC3Ⅱ、P62、Beclin-1mRNA表达显著降低(P<0.05);与其他各干预组比较,活血定眩胶囊含药血清中剂量组上述指标改善较明显(P<0.05)。结论:活血定眩胶囊含药血清可通过调节血管内皮细胞适度自噬而减轻内皮细胞损伤,是活血定眩胶囊治疗椎动脉型颈椎病的机制之一。Objective: To investigate the effects of serum containing Huoxue Dingxuan Capsule on autophagy of bEnd.3 cells cultured in oxygen-glucose deprivation(OGD). Methods: The drug-containing serum of rats was prepared, and the OGD model was established by the subculture of bEnd.3 cells. The cells were divided into the model group, Huoxue Dingxuan Capsule drug-containing serum low, medium and high dose groups and flunarizine hydrochloride group. The bEnd.3 cells were used as the control group. The corresponding methods were used to intervene. After the intervention, the MDA concentration and LD activity of the cells in each group were detected. Confocal laser scanning microscopy was used to analyze LC3 punctate aggregation count and calculate the gray value. the qRT-PCR method was used to detect the expression of autophagy-related genes LC3 Ⅱ, P62, Beclin-1 mRNA. Results: Compared with the control group, the MDA concentration, LD activity, LC3 Ⅱ expression and the expression of LC3 Ⅱ, P62 and Beclin-1 mRNA in the OGD model group were increased(P<0.05);compared with the OGD model group, the MDA concentration, LD activity and the expression of LC3 Ⅱ, and the mRNA expression of LC3 Ⅱ, P62 and Beclin-1 were decreased in each group after the intervention of Huoxue Dingxuan Capsule containing serum medium and high dose groups(P<0.05);compared with other intervention groups, the above indicators were significantly improved in the Huoxue Dingxuan Capsule containing serum medium dose group(P<0.05). Conclusion: The serum containing Huoxue Dingxuan Capsule can alleviate the injury of vascular endothelial cells by regulating moderate autophagy, which is one of the mechanisms of Huoxue Dingxuan Capsule in treating CSA.

关 键 词：活血定眩胶囊 含药血清 bEnd.3细胞 自噬 脑微血管 内皮细胞 椎动脉型颈椎病 机制 

分 类 号：R285.5[医药卫生—中药学]