当归多糖促进大鼠心肌细胞系H9c2增殖

基础医学与临床 ›› 2021, Vol. 41 ›› Issue (4): 551-557.

当归多糖促进大鼠心肌细胞系H9c2增殖

杨萍, 代天, 张苏川^*

江汉大学附属医院(武汉市第六医院)心血管内科, 湖北武汉 430015

收稿日期:2020-04-06 修回日期:2020-08-11 出版日期:2021-04-05 发布日期:2021-04-05
通讯作者: ^*zhangsuchuan1@163.com

Angelica polysaccharide promotes the proliferation of rat cardiomyocyte line H9c2

YANG Ping, DAI Tian, ZHANG Su-chuan^*

Department of Cardiovascular Medicine, the Affiliated Hospital of Jianghan University (Wuhan Sixth Hospital),Wuhan 430015, China

Received:2020-04-06 Revised:2020-08-11 Online:2021-04-05 Published:2021-04-05
Contact: ^*zhangsuchuan1@163.com

摘要/Abstract

摘要： 目的研究当归多糖(APS)对大鼠心肌细胞系H9c2增殖和凋亡的影响。方法将大鼠心肌细胞系H9c2分为对照组(control组)、阿霉素(ADR)诱导H9c2细胞复制扩张性心肌病心肌细胞组(ADR组)、APS干预(APS组)、转染anti-miR-NC(APS+anti-miR-NC组)及转染anti-miR-4701-3p(APS+anti-miR-4701-3p组)。采用流式细胞计量术检测细胞凋亡,Western blot检测B细胞淋巴瘤/白血病-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、细胞核相关抗原Ki-67(Ki-67)、增殖细胞核抗原(PCNA)蛋白表达,酶联免疫(ELISA)法检测B型脑钠肽 (BNP) 水平,细胞计数试剂盒8(CCK-8)法检测细胞的增殖,实时定量PCR(RT-qPCR)检测微小RNA-4701-3p(miR-4701-3p)表达。结果阿霉素明显提高H9c2细胞的BNP含量、细胞凋亡率、Bax蛋白水平,显著降低Bcl-2蛋白表达量(P＜0.05)。当归多糖明显增加阿霉素诱导后H9c2细胞的细胞活力、Ki-67、PCNA、Bcl-2蛋白表达量和miR-4701-3p表达量,显著减少细胞的凋亡率、Bax蛋白表达量(P＜0.05)。敲减miR-4701-3p明显降低当归多糖作用的扩张性心肌病H9c2细胞的细胞活力、Ki-67、PCNA、Bcl-2蛋白水平,而显著提高其凋亡率、Bax蛋白表达量(P＜0.05)。结论当归多糖通过上调miR-4701-3p的表达,促进阿霉素诱导的扩张性心肌病大鼠心肌细胞系H9c2增殖。

关键词: 阿霉素, 细胞系H9c2, miR-4701-3p, B型脑钠肽

Abstract: Objective To study the effect of angelica polysaccharide on proliferation and apoptosis of rat cardiomyocyte line H9c2. Methods Rat cardiomyocyte H9c2 was divided into a control group (Control group), doxorubicin (ADR) induced H9c2 cell replication dilated cardiomyopathy cardiomyocyte group (ADR group), APS intervention (APS group), transfection anti-miR-NC (APS+anti-miR-NC group) and transfected anti-miR-4701-3p(APS+anti-miR-4701-3p group). Flow cytometry was applied to determine apoptosis, Western blot was employed to test B cell lymphoma/leukemia-2(Bcl-2), Bcl-2 related X protein (Bax), nuclear related antigen Ki-67(Ki-67) and proliferation cell nuclear antigen (PCNA) protein expression, enzyme-linked immunoassay (ELISA) method was used to detect B-type brain natriuretic peptide (BNP) level. Cell proliferation was detected by commercially available kit 8(CCK-8), and real-time quantitative PCR (RT-qPCR) was used to analyze the microRNA-4701-3p(miR-4701-3p) expression. Results Adriamycin significantly increased BNP content, apoptosis rate, and Bax protein level in H9c2 cells, and greatly reduced Bcl-2 protein expression(P＜0.05). Angelica polysac-charide obviously increased H9c2 cell viability, Ki-67, PCNA, Bcl-2 protein expression and miR-4701-3p expression after doxorubicin induction, and remarkably reduced cell apoptosis and Bax protein expression(P＜0.05). Knockdown of miR-4701-3p significantly reduced the cell viability, Ki-67, PCNA, and Bcl-2 protein levels of dilated cardiomyopathy H9c2 cells treated with angelica polysaccharide, while significantly increased its apoptosis rate and Bax protein expression(P＜ 0.05). Conclusions Angelica polysaccharide up-regulates the expression of miR-4701-3p, promotes the proliferation of adriamycin-induced rat cardiomyocyte line H9c2.

Key words: adriamycin, cell line H9c2, miR-4701-3p, BNP

中图分类号:

R542.2

杨萍, 代天, 张苏川. 当归多糖促进大鼠心肌细胞系H9c2增殖[J]. 基础医学与临床, 2021, 41(4): 551-557.

YANG Ping, DAI Tian, ZHANG Su-chuan. Angelica polysaccharide promotes the proliferation of rat cardiomyocyte line H9c2[J]. Basic & Clinical Medicine, 2021, 41(4): 551-557.

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