白藜三醇通过下调microRNA-21减轻快速电刺激所致心房肌细胞电重构  

Resveratrol reduces electrical remodeling in atrial fibrillation by downregulating microRNA-21 in neonatal rat atrial myocytes

作　　者：张松 沈冰冰 李飞[2] 朱启仁 王志荣 张卓琦 ZHANG Song;SHEN Bing-bing;LI Fei;ZHU Qi-ren;WANG Zhi-rong;ZHANG Zhuo-qi(Department of Cardiology, The Affiliated Hospital of Xuzhou Medical College, Xuzhou 221002, China;Pizhou People's Hospital, Pizhou 221300, China)

机构地区：[1]徐州医科大学附属医院心内科,江苏徐州221002 [2]邳州市人民医院,江苏邳州221300

出　　处：《中国病理生理杂志》2017年第8期1353-1358,共6页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30800219);徐州市医学青年后备人才项目

摘　　要：目的:研究白藜三醇(resveratrol,RSV)对快速电刺激(rapid electrical stimulation,RES)导致乳鼠心房肌细胞电重构时微小RNA-21(microRNA-21,miR-21)表达的影响,探讨RSV通过miR-21参与电重构的可能机制。方法:采用胰酶、Ⅰ型胶原酶双酶法及差速贴壁法分离培养乳鼠心房肌细胞。通过RES建立乳鼠心房肌细胞房颤模型,心房肌细胞随机分为4组:空白对照(control)组、RSV组、RES组和RSV+RES组。为了证实RSV是否通过调控miR-21的表达参与电重构,除了上述4组,另增加过表达和沉默miR-21组:RES+阴性对照组(RES+NC组)、RES+miR-21 mimics组、RES+miR-21 mimics+RSV组、RES+miR-21 inhibitor组和RES+miR-21 inhibitor+RSV组。CCK-8法检测心房肌细胞活性以确定RSV最佳作用浓度及时间,q PCR法检测各组细胞内miR-21及L型钙离子通道CACNA1C、CACNB2 mRNA的表达水平,Western blot检测L型钙离子通道Cav1.2和Cavβ_2 的蛋白表达水平。结果:与control组相比,RES组miR-21表达明显上调(P<0.05),加入RSV预处理后miR-21表达下调(P<0.05)。与RES+miR-21 mimics组相比,RES+miR-21 mimics+RSV组miR-21表达下调(P<0.05),而CACNA1C和CACNB2 mRNA及Cav1.2和Cavβ_2 蛋白表达量增加(P<0.05)。与RES组比,RES+miR-21 inhibitor和RES+miR-21 inhibitor+RSV组的miR-21表达下调(P<0.05),CACNA1C和CACNB2 mRNA及Cav1.2和Cavβ_2 蛋白表达量增加,但RES+miR-21 inhibitor组与RSV+RES组比,miR-21表达、CACNA1C和CACNB2 mRNA及Cav1.2和Cavβ_2 蛋白表达量的差异无统计学显著性。结论:在快速电刺激乳鼠心房肌细胞模拟房颤模型中,RSV干预可能通过下调miR-21表达而调控其下游靶基因这一途径来减轻心房肌细胞电重构。AIM:To detect the effects of resveratrol(RSV)on the expression of microRNA-21(m iR-21)inprimarily cultured neonatal rat atrial myocytes with electric remodeling induced by rapid electrical stimulation(RES).Furthermore,to find out the possible mechanism of miR-21regulating electrical remodeling.METHODS:The neonatal rat atrialmyocytes were isolated by double-enzyme(trypsin and collagenase I)digestion and differential adhesion method.Theatrial fibrillation(AF)model was induced by RES.Atrial myocytes were randomly divided into4groups:control group,RSV group,RES group,and RSV+RES group.To further detect whether RSV regulated electric remodeling by miR-21,except the4groups,we add miR-21over-expression group and miR-21inhibitor group:RES+negative control(NC)group,RES+miR-21mimics group,RES+miR-21mimics+RSV group,RES+miR-21inhibitor group,and RES+miR-21inhibitor+RSV group.The optimal concentration and pretreatment time of resveratrol were determined by CCK-8assay.The expression of miR-21and the mRNA expression of L-type calcium channels CACNA1C and CACNB2in atrial myocyteswere detected by qPCR.The protein expression of L-type calcium channels C avl.2and Cavp2in the atrial myocytes was analyzed by Western blot.RESULTS:The expression of miR-21in RES group was significantly increased compared withcontrol group,while preconditioning with RSV decreased the expression of miR-21.Compared with RES+miR-21mimicsgroup,the expression of miR-21in RES+miR-21mimics+RSV group was significantly decreased.Meanwhile,the mRNAexpression of CACNA1C and CACNB2,and the protein levels of C avl.2and Cavp2were increased(P<0.05).Comparedwith RES group,the expression of miR-21in RES+miR-21inhibitor group and RES+miR-21inhibitor+RSV group wasdecreased,while the mRNA expression of CACNA1C and CACNB2,and the protein levels of C avl.2and Cavp2were increased.However,no difference of the expression of miR-21,the mRNA expression of CACNA1C and CACNB2,and theprotein levels of C avl.2and Cavp2among RSV+RES,RES+miR-21inhibitor and RES+miR-21inhibitor+RSV grou

关 键 词：白藜三醇 微小RNA-21 快速电刺激 电重构 乳鼠心房肌细胞 

分 类 号：R541.7[医药卫生—心血管疾病] R965[医药卫生—内科学]