当归多糖促进大鼠心肌细胞系H9c2增殖

Angelica polysaccharide promotes the proliferation of rat cardiomyocyte line H9c2

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。

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 increasedH9c2 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.