丙泊酚调控miR-21-3p对缺氧/复氧所诱导心肌细胞损伤的影响

Effect of propofol on hypoxia/reoxygenation-induced cardiomyocyte injury by regulating miR-21-3p

目的探讨丙泊酚对缺氧/复氧(hypoxia/reoxygenation,H/R)诱导的心肌细胞损伤的影响及其可能作用机制。方法以H/R诱导的大鼠心肌细胞H9C2建立心肌缺血再灌注损伤模型,不同剂量的丙泊酚处理H9C2细胞;采用CCK-8法、流式细胞术分别检测细胞增殖及凋亡;采用qRT-PCR法检测miR-21-3p的表达量;ELISA法检测白细胞介素6(interleukin-6,IL-6)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-1β(interleukin-1β,IL-1β)的水平;anti-miR-NC、anti-miR-21-3p分别转染至H9C2细胞后进行H/R处理,miR-NC、miR-21-3p mimics分别转染至H9C2细胞后用50μmol/L丙泊酚处理24 h,然后进行H/R处理,采用上述方法分别检测细胞增殖、凋亡及炎性细胞因子表达水平;Western blot测定B淋巴细胞瘤-2相关蛋白(Bcl-2-associated X protein,Bax)、B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)蛋白表达量。结果丙泊酚处理后,H/R诱导的H9C2细胞增殖抑制率、TNF-α、细胞凋亡率、IL-1β、Bax蛋白水平、IL-6、miR-21-3p表达量下降(P<0.05),Bcl-2蛋白水平上升(P<0.05),且呈剂量依赖性;转染anti-miR-21-3p可降低细胞增殖抑制率、细胞凋亡率、Bax蛋白水平、IL-6、TNF-α、IL-1β水平(P<0.05),Bcl-2蛋白水平升高(P<0.05);转染miR-21-3p mimics可减弱丙泊酚对H/R诱导的H9C2细胞增殖、凋亡及炎性因子表达水平的作用。结论丙泊酚可通过抑制miR-21-3p表达而促进细胞增殖及抑制细胞凋亡、炎性反应从而减轻H/R诱导的心肌细胞损伤。

Objective To investigate the effect of propofol on hypoxia/reoxygenation(H/R)-induced myocardial cell injury and its possible mechanism.Methods The myocardial ischemia-reperfusion injury model was established in H/R-induced rat cardiomyocytes H9C2,which were later treated with different doses of propofol.Cell proliferation and apoptosis were detected by CCK-8 assay and flow cytometry,respectively.The expression of miR-21-3p was detected by Quantitative real-time polymerase chain reaction(QRT-PCR).The levels of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)were detected by Enzyme-linked immunosorbent assay(ELISA).Anti-miR-NC and anti-miR-21-3p were transfected into H9C2 cells,followed by H/R induction.miR-NC and miR-21-3p mimics were transfected into H9C2 cells,followed by 24-h treatment of 50μmol/L propofol and H/R induction.Cell proliferation,apoptosis and the expression levels of inflammatory cytokines were detected.Western blot was used to detect the expressions of Bax,and Bcl-2.Results Propofol treatment dose-dependently reduced anti-proliferative rate,apoptotic rate,and expression levels of TNF-α,IL-1β,IL-6,miR-21-3p and Bax in H/R-induced H9C2 cells,while up-regulated protein level of Bcl-2(P<0.05).Transfection of anti-miR-21-3p reduced anti-proliferative rate,apoptotic rate,and expression levels of TNF-α,IL-1β,IL-6,and Bax,and up-regulated Bcl-2(P<0.05).Transfection of miR-21-3p mimics attenuated the regulatory effect of propofol on the proliferation,apoptosis and expression levels of inflammatory factors in H/R-induced H9C2 cells.Conclusion Propofol could promote cell proliferation and inhibit apoptosis and inflammatory response by inhibiting the expression of miR-21-3p,thereby reducing H/R-induced cardiomyocyte injury.