摘要
目的:研究丹蛭降糖胶囊对高糖诱导的人脐静脉内皮细胞(HUVECs)细胞凋亡及线粒体凋亡途径与死亡受体凋亡途径的影响,探讨丹蛭降糖胶囊的血管保护作用机制。方法:体外培养HUVECs细胞株,MTT法筛选丹蛭降糖胶囊含药血清对HUVECs最佳浓度范围。高糖诱导HUVECs细胞损伤模型,采用MTT法检测细胞存活率,Hoechst 33258染色观察细胞凋亡形态,流式细胞术检测细胞凋亡率,荧光定量PCR法检测细胞内Bax、Bcl-2、Cyt-c、cleaved-caspase-9、Fas、cleaved-caspase-8、cleaved-caspase-3 mRNA表达。结果:高糖培养导致HUVECs存活率下降(P<0.01),细胞凋亡形态表现明显,细胞凋亡率增加(P<0.01),Bax、Cyt-c、cleaved-caspase-9、Fas、cleaved-caspase-8、cleaved-caspase-3 mRNA表达上调(P<0.01),Bcl-2 mRNA表达下调(P<0.01);丹蛭降糖胶囊含药血清低、中、高剂量组细胞存活率显著上调(P<0.01,P<0.05),细胞凋亡形态改变减少,细胞凋亡率减少(P<0.01,P<0.05);低、中剂量组Bax、Cyt-c、cleaved-caspase-9、cleaved-caspase-3 mRNA表达显著下调(P<0.01),Bcl-2 mRNA表达显著上调(P<0.01);高剂量组Bax、Cyt-c、cleaved-caspase-9、Fas、cleavedcaspase-8、cleaved-caspase-3 mRNA表达显著下调(P<0.01),Bcl-2 mRNA表达显著上调(P<0.01)。结论:丹蛭降糖胶囊含药血清对高糖诱导的HUVECs损伤具有改善作用,能减少细胞凋亡,其机制可能与抑制线粒体介导的内部凋亡途径与死亡受体介导的外部凋亡途径有关,且浓度不同抑制的凋亡途径有差异。
Objective: To study the effects of Danzhi Jiangtang Capsule(DJC) on apoptosis and related pathways of death receptors apoptotic and mitochondrial apoptotic of human umbilical vein endothelial cells(HUVECs) induced by high glucose, and to investigate the vascular protective mechanism of DJC. Methods: The HUVECs cell line was cultured in vitro,and the optimal concentration range of HUVECs was determined by MTT method. High glucose induced HUVECs cell injury model, and MTT assays were used to detect cell survival rate, apoptosis morphology was observed by Hoechst 33258 staining,cell apoptosis rate was assayed by flow cytometry, and mRNA expressions of Bcl-2, Bax, Cyt-c, cleaved-caspase-9, Fas, cleavedcaspase-8, cleaved-caspase-3 in cell was detected by PCR of fluorescence quantitative. Results: Compared with the normal group,the survival rate of HUVECs in the model group decreased(P<0.01), the morphology of apoptosis was obvious, the apoptosis rate increased(P<0.01), and the mRNA expressions of Bax, Cyt-c, cleaved-caspase-9, Fas, cleaved-caspase-8 and cleaved-caspase-3 were up-regulated(P<0.01). The mRNA expression of Bcl-2 was down-regulated(P<0.01). Compared with the model group,the survival rate of the groups of DJC-containing serum were significantly up-regulated(P<0.01, P<0.05), the morphology of apoptosis was decreased, and the apoptosis rate were decreased(P<0.01, P<0.05). The mRNA expressions of Bax, Cyt-c, cleavedcaspase-9 and cleaved-caspase-3 of the groups of low-concentration and medium-concentration DJC-containing serum were downregulated(P<0.01), and mRNA expression of Bcl-2 was up-regulated(P<0.01). The mRNA expressions of Bax, Cyt-c, cleavedcaspase-9, Fas, cleaved-caspase-8 and cleaved-caspase-3 of the group of high-concentration DJC-containing serum were downregulated(P<0.01), and the mRNA expression of Bcl-2 was up-regulated(P<0.01). Conclusion: The drug-containing serum of DJC can improve the damage of high glucose-induced HUVECs, inhibit the excessive activation apoptosis, suppress the pathways o
作者
尤良震
吴袁元
尹昀东
刘蒙蒙
于东东
申国明
方朝晖
YOU Liang-zhen;WU Yuan-yuan;YIN Yun-dong;LIU Meng-meng;YU Dong-dong;SHEN Guo-ming;FANG Zhao-hui(Graduate School,Anhui University of Chinese Medicine,Hefei 230031,China;Anhui Academic of Traditional Chinese Medicine Diabetes Research Institute,Hefei 230031,China;Endocrine Department,The First Affiliated Hospital of Anhui University of Chinese Medicine,Hefei 230031,China)
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2019年第8期3675-3680,共6页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
国家中医药管理局中医临床基地科研专项(No.JDZX2015001)
国家自然科学基金面上项目(No.81774286,No.81573944)~~
关键词
丹蛭降糖胶囊
糖尿病
人脐静脉内皮细胞
细胞凋亡
死亡受体凋亡途径
线粒体凋亡途径
Danzhi Jiangtang Capsule
Diabetes mellitus
Human umbilical vein endothelial cells(HUVECs)
Apoptosis
Death receptors apoptotic pathway
Mitochondrial apoptotic pathway
