活血胶囊激活PI3K/Akt/Nrf2/HO-1通路对血管内皮细胞发挥抗氧化损伤作用  

HXJN Mitigate Oxidative Stress in Brain Microvascular Endothelial Cells via Regulation of PI3K/Akt/Nrf2/HO-1 Pathways

作　　者：赵静[1] 袁瑞华 王海芳 邓国荣 张琳萍[3] 曹情雯 霍雪萍[5] 赵向绒[5] 刘勤社[1] ZHAO Jing;YUAN Ruihua;WANG Haifang;DENG Guorong;ZHANG Linping;CAO Qingwen;HUO Xueping;ZHAO Xiangrong;LIU Qinshe(Department of Integrated Traditional Chinese and Western Medicine for Cardiovascular Diseases,Shaanxi Key Laboratory of Integrated Traditional and Western Medicine for Prevention and Treatment of Cardiovascular Diseases,Shaanxi University of Traditional Chinese Medicine,Xi'an 712046,China;Department of Cardiology,the Second Affiliated Hospital of Shaanxi University of Traditional Chinese Medicine,Xi'an 712000,China;Department of Integrated Traditional Chinese and Western Medicine,Medical School of Xi'an Jiaotong University,Xi'an 710061,China;Medical Management Office,Shaanxi Province Hospital of Chinese Medicine,Xi'an 710003,China;Central Laboratory of Shaanxi Provincial People's Hospital,Xi'an 710068,China)

机构地区：[1]陕西中医药大学中西医结合心血管病专业,陕西省中西医结合心血管病防治重点实验室,陕西西安712046 [2]陕西中医药大学第二附属医院心内科,陕西西安712000 [3]西安交通大学医学部中西医结合心血管专业,陕西西安710061 [4]陕西省中医医院医疗管理处,陕西西安710003 [5]陕西省人民医院中心实验室,陕西西安710068

出　　处：《云南中医学院学报》2019年第3期10-16,共7页Journal of Yunnan University of Traditional Chinese Medicine

基　　金：国家自然科学基金(81573823);陕西省中医管理局平台项目(JCPT028);陕西省中医管理局项目(2019-ZZ-JC025)。

摘　　要：目的研究活血胶囊(HXJN)对血管内皮细胞的抗氧化损伤作用分子机制。方法采用WST-1法检测细胞活力;qRT-PCR和Western blot技术检测血红素加氧酶-1(HO-1)的表达;检测HXJN对Akt和转录因子Nrf2的激活作用,并分析其对HO-1表达的影响。结果HXJN可显著减轻H2O2对细胞活力的抑制作用。HXJN可上调HO-1表达水平。HXJN可诱导Akt和Nrf2磷酸化,并促进Nrf2核转位;LY294002、perifosine和ML385均抑制HXJN对HO-1的上调,LY294002可抑制Nrf2核转位。结论HXJN促进HO-1表达而发挥抗氧化损伤作用,Akt和Nrf2参与调控HO-1表达。Objective To investigate the anti-oxidant effects of HXJN in brain microvascular endothelial cells(bEND.3)and the underlying mechanism.Methods HXJN-containing serum and HXJN extract solution were prepared and their effects on bEND.3 cell viability were measured by WST-1 assay.The protective effect of HXJN on H2O2-induced oxidative cell damage was also measured.Heme oxygenase(HO-1)mRNA level was detected by quantitative RT-PCR.The expression levels of HO-1,ph-Akt,ph-Nrf2(nuclear factor erythroid 2,Nrf2)and Nrf2 nuclear translocation were detected by using Western Blot analysis.The involvement of Akt/Nrf2/HO-1 signaling pathway in the effect of HXJN were analyzed using specific pharmacological inhibitors.Results Treatment of the bEND.3 cells with HXJN-containing serum less than 10%for 24 h did not reduce the cell viability.Treatment of the cells with HXJN extract for 24 h increased the cell viability significantly.The cell viability was greatly decreased by a 3 h's treatment of 200μM or 300μM H2O2,and HXJN showed a significant protective effect on the cell injury induced by 200μM H2O2(P<0.01).Furthermore,HXJN induced the expression of HO-1 and increased the phosphorylation and nuclear translocation of Nrf2.HXJN-induced HO-1 up-regulation was suppressed by LY294002,perifosine and ML385.The nuclear translocation of Nrf2 was inhibited by LY294002.Conclusion HXJN protects bEND.3 against oxidative stress through up-regulating the expression of HO-1.PI3K/Akt/Nrf2 signaling pathway may be involved in this effect.

关 键 词：活血胶囊 血管内皮细胞 Nrf-2 HO-1 氧化损伤 

分 类 号：R285.5[医药卫生—中药学]