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汪晶, 谭立业, 刘卓, 韩莹. miR-221在急性心肌梗死大鼠中表达及其作用[J]. 中国公共卫生, 2017, 33(6): 930-933. DOI: 10.11847/zgggws2017-33-06-16
引用本文: 汪晶, 谭立业, 刘卓, 韩莹. miR-221在急性心肌梗死大鼠中表达及其作用[J]. 中国公共卫生, 2017, 33(6): 930-933. DOI: 10.11847/zgggws2017-33-06-16
WANG Jing, TAN Li-ye, LIU Zhuo.et al, . Expression and role of miR-221 in rats with acute myocardial infarction[J]. Chinese Journal of Public Health, 2017, 33(6): 930-933. DOI: 10.11847/zgggws2017-33-06-16
Citation: WANG Jing, TAN Li-ye, LIU Zhuo.et al, . Expression and role of miR-221 in rats with acute myocardial infarction[J]. Chinese Journal of Public Health, 2017, 33(6): 930-933. DOI: 10.11847/zgggws2017-33-06-16

miR-221在急性心肌梗死大鼠中表达及其作用

Expression and role of miR-221 in rats with acute myocardial infarction

  • 摘要: 目的 探讨miR-221在急性心肌梗死(AMI)大鼠中的表达及其作用。方法 在45只SD清洁级雄性大鼠中随机抽取35只通过冠状动脉左前降支结扎大鼠建立AMI模型,将其中建模成功的30只大鼠随机分为AMI组、对照组及抑制组,每组各10只;另抽取未建模的10只大鼠设立假手术组,采用冠状动脉左前降支穿线不结扎。其中对照组用携带绿色荧光蛋白(GFP)的慢病毒空载体miR-221阴性对照以4×107病毒数量进行心肌组织局部注射转染,抑制组用携带miR-221 inhibitor慢病毒以4×107病毒数量进行心肌组织局部注射转染,AMI组和假手术组每天给予等量生理盐水,隔日1次连续2周;记录大鼠心脏功能。采用原位末端凋亡法(TUNEL)检测心肌凋亡指数(AI),RT-PCR法检测miR-221表达,Western blot法检测Bax、Bcl-2、PI3K和p-AKT蛋白表达。结果 与假手术组大鼠miR-221表达量(0.18±0.02)比较,AMI组(1.16±0.12)和对照组(1.18±0.12)大鼠的miR-221表达量均升高(均P<0.01),抑制组大鼠miR-221表达量(0.30±0.03)均低于AMI组和对照组大鼠的miR-221表达量(均P<0.01);与假手术组大鼠Bax(0.13±0.01)、Bcl-2(0.53±0.05)、PI3K(0.45±0.04)和p-AKT(0.87±0.09)蛋白表达量比较,AMI组和对照组大鼠的Bax蛋白表达量均上调(均P<0.01),Bcl-2、PI3K和p-AKT蛋白表达量均下调(均P<0.01);抑制组大鼠较AMI组和对照组大鼠Bax表达量均下调(P<0.01),Bcl-2、PI3K和p-AKT 蛋白表达量均上调(P<0.01)。结论 miR-221在AMI大鼠心肌组织中高表达,下调miR-221表达可通过激活PI3K/AKT信号通路调控下游蛋白Bax及Bcl-2的表达抑制AMI大鼠心肌细胞凋亡。

     

    Abstract: Objective To explore the expression and role of microRNAs-221 (miR-221) in rats with acute myocardial infarction (AMI).Methods A total of 35 male Sprague-Dawley (SD) rats were randomly selected from 45 rats to establish an AMI model by ligation of left anterior descending branch of coronary artery;then 30 of the model rats with AMI successfully established were randomly divided into an AMI,a control and an inhibition group (10 in each group);another 10 rats of non-model group were regarded as a sham operation group without the ligation.The rats of control group were transfected with miR-221 negative non-carrier lentiviral vector marked by green fluorescent protein (GFP) at the dosage of 4 x 107 viruses via injection into myocardial tissues.The rats of inhibition group were treated the same as those of control group but with miR-221 inhibitor positive lentiviral vector.The rats of AMI and sham operation group were injected with normal saline.All the treatments were performed once the every other day consecutively for two weeks.T cardiac function of all the rats was recorded during the treatment period.The expression of miR-221 in myocardial was detected with reverse transcription-PCR (RT-PCR).Apoptosis index (AI) of myocardium was calculated based on the results of TdT-mediated dUTP-biotin nick end-labeling (TUNEL) assay.The expression of Bcl-2 (B-cellymphoma-2),Bax (B-cell lymphoma 2-associated X protein),PI3K (phosphoinositide 3-OH kinase),and p-AKT (phosphorylated protein kinase B) were determined with Western blot.Results Compared with that of sham operation group (0.18±0.02),the expression of miR-221 in AMI group (1.16±0.12) and control group (1.16±0.12) increased significantly (both P<0.01).The expression of miR-221 (0.30±0.03) in the inhibition group was lower than that in AMI group and control group (both P<0.01).Compared with the expression of Bax (0.13±0.01),Bcl-2 (0.53±0.05),PI3K (0.45±0.04),and p-AKT (0.87±0.09) in sham operation group,the expression of Bax increased;the expression of Bcl-2,PI3K,and p-AKT decreased in AMI group and control group (P<0.01 for all).The expression of Bax decreased;while the expression of Bcl-2,PI3K and p-AKT increased significantly in the inhibition group (all P<0.01).Conclusion The expression of miR-221 is high in the myocardium of rats with AMI and down-regulated miR-221 expression could inhibit the apoptosis of myocardium in rats with AMI by activating PI3K/AKT signaling pathway to regulate the expression of the downstream protein of Bax and Bcl-2.

     

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