摘要
目的探讨四逆汤含药血清在减轻大鼠心肌细胞损伤中,微小RNA-21(miR-21)靶向调控程序性细胞死亡因子4(PDCD4)的意义。方法选择SD大鼠制备四逆汤含药血清,体外培养大鼠心肌细胞(H9C2),通过RNA干扰技术,转染miR-21 mimic及miR-21 inhibitor,建立稳定的miR-21 mimic和miR-21 inhibitor细胞系。细胞分别给予阿霉素(ADR)10μg/mL作用24 h损伤和无ADR处理,再各分为四个水平,分别为miR-21 mimic、miR-21 inhibitor、四逆汤+miR-21mimic、四逆汤+miR-21 inhibitor,另设正常对照组及四逆汤组,共10组。采用实时荧光定量PCR法检测各组细胞miR-21、PDCD4表达,采用流式细胞术检测各组心肌细胞凋亡百分率。结果与正常对照组相比,ADR+miR-21 mimic组、ADR+miR-21 inhibitor组、ADR+四逆汤+miR-21 mimic组、ADR+四逆汤+miR-21 inhibitor组细胞凋亡百分率、PDCD4表达量增加,而miR-21的表达量下降(P均<0.05或0.01);ADR+miR-21 mimic组相比,ADR+四逆汤+miR-21 mimic组miR-21表达上升,PDCD4下降,凋亡百分率降低(P均<0.05);与ADR+miR-21 inhibitor组相比,ADR+四逆汤+miR-21 inhibitor组细胞miR-21表达上升,PDCD4下降,凋亡百分率降低(P均<0.05)。结论四逆汤可抑制阿霉素损伤大鼠心肌细胞凋亡,减轻心肌细胞损伤,其机制可能与上调miR-21的表达靶向调控PDCD4有关。
Objective To investigate the significance of microRNA-21(miR-21)in the targeted regulation of programmed cell death factor 4(PDCD4)in the treatment of myocardial cell injury in rats with Sini decoction-containing serum.Methods Sprague Dawley(SD)rats were selected to the prepare Sini decoction-containing serum,and the rat myocardial cells H9C2 were cultured in vitro.After transfection with miR-21 mimic and miR-21 inhibitor by RNA interference technique,stable miR-21 mimic and miR-21 inhibitor cell lines were established.The cells were separately given adriamycin(ADR)injury(10μg/mL)for 24 h and without ADR treatment,and then each was divided into four levels,respectively:the miR-21 mimic group,miR-21 inhibitor group,Sini decoction+miR-21 mimic group,and Sini decoction+miR-21 inhibitor group,meanwhile,the Sini decoction group and normal control group were set,a total of 10 groups.We used real-time fluorescent quantitative PCR method to detect miR-21 and PDCD4 expression,and the percentage of cardiomyocyte apoptosis in each group was detected by flow cytometry.Results Compared with the normal control group,the apoptosis rate and PDCD4 expression of the ADR+miR-21 mimic group,ADR+miR-21 inhibitor group,ADR+Sini decoction+miR-21mimic group,and ADR+Sini decoction+miR-21 inhibitor group increased significantly,while the expression of miR-21 decreased,with statistically significant differences(P<0.05,or P<0.01).Compared with the ADR+miR-21 mimic group,the expression of miR-21 increased,and PDCD4 decreased,and the percentage of apoptosis decreased in the ADR+Sini decoction+miR-21 mimic group(all P<0.05).Compared with the ADR+miR-21 inhibitor group,the expression of miR-21 increased,but the PDCD4 expression decreased,and the percentage of apoptosis decreased in the ADR+Sini decoction+miR-21 inhibitor group,with statistically significant differences(all P<0.05).Conclusion Sini decoction can inhibit apoptosis and alleviate myocardial cell injury in ADR-injured rats,and its mechanism may be related to the targeted regulation of miR-21 expression in PDCD4.
作者
梁丽英
刘欢
张永琴
陈炜
陈晶
LIANG Liying;LIU huan;ZHANG Yongqin;CHEN Wei;CHEN Jing(The First Affiliated Hospital of Guangxi University of Traditional Chinese Medicine,Nanning 530023,China)
出处
《山东医药》
CAS
2019年第23期14-18,共5页
Shandong Medical Journal
基金
广西自然科学基金资助项目(2016GXNSFBA380018)
广西高校中医临床研究重点实验室课题(J16066)
广西中医基础研究重点实验室开放研究课题(KJT1701207)
广西中医药大学岐黄工程高层次人才团队培育项目(2018003)
