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(PDF)

《云南中医学院学报》[ISSN:1000-2723/CN:53-1048/R]

期数:

2019年03期

页码:

10-16

栏目:

实验研究

出版日期:

2020-04-20

文章信息/Info

Title:

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

文章编号:

1000-2723（2019）03-0010-07

作者:

赵 静¹; 袁瑞华¹; 王海芳¹; 邓国荣²; 张琳萍³; 曹情雯⁴; 霍雪萍⁵; 赵向绒⁵; 刘勤社^1△

（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）

Author(s):

ZHAO Jing¹;  YUAN Ruihua¹;  WANG Haifang¹;  DENG Guorong²;  ZHANG Linping³;  CAO Qingwen⁴;  HUO Xueping⁵;  ZHAO Xiangrong⁵;  LIU Qinshe¹

(1. 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；2. Department of Cardiology, the Second Affiliated Hospital of Shaanxi University of Traditional Chinese Medicine, Xi’an 712000, China;）

关键词:

活血胶囊; 血管内皮细胞; Nrf-2; HO-1; 氧化损伤

Keywords:

Huoxue Capsule (HXJN);  brain microvascular endothelial cells;  nuclear factor E2 related factor 2;  heme oxygenase-1;  oxidative injury

分类号:

R285.5

DOI:

10.19288/j.cnki.issn.1000-2723.2019.03.002

文献标识码:

A

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。

Abstract:

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 200 μM or 300 μM H2O2, and HXJN showed a significant protective effect on the cell injury induced by 200 μM 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.

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备注/Memo

备注/Memo:

收稿日期： 2019 - 03- 10
基金项目： 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）
第一作者简介： 赵静（1990- ），女，在读硕士研究生，研究方向：中西医结合心血管病防治。
△通信作者： 刘勤社，E-mail：lqsspph@126.com

作者单位：3.Department of Integrated Traditional Chinese and Western Medicine, Medical School of Xi’an Jiaotong University, Xi’an 710061, China;

　　4.Medical Management Office, Shaanxi Province Hospital of Chinese Medicine, Xi’an 710003, China;?/div>

　　5.Central Laboratory of Shaanxi ProvincialPeople’s Hospital, Xi’an 710068, China)

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