生长分化因子15过表达对心肌缺血再灌注的保护作用及与其自噬的关系

发布时间：2018-03-12 14:15

  本文选题：生长分化因子　切入点：慢病毒感染　出处：《中国临床药理学杂志》2017年10期 　论文类型：期刊论文

【摘要】：目的通过构建生长分化因子15(GDF15)稳定过表达的心肌细胞株,探讨其对H9c2心肌细胞缺血/再灌注(I/R)损伤凋亡和自噬的影响。方法建立H9c2 I/R损伤模型,用靶向GDF15过表达慢病毒和阴性空载体病毒转染H9c2细胞,筛选出GDF15稳定过表达细胞株。分为3组:对照组、空载体组和GDF15过表达组。用MTT法检测I/R后细胞存活率,以LDH试剂盒检测细胞培养上清液中乳酸脱氢酶活力,以免疫印迹法检测凋亡相关蛋白Caspase-3、Bcl-2、Bax和自噬相关蛋白Beclin-1、LC3-B的表达。结果 I/R 2 h,对照组、空载体组和过表达组的细胞存活率分别为(60.47±5.03)%,(63.21±17.13)%,(74.93±10.03)%,过表达组细胞活性较对照组与空载体组均增高,差异有统计学意义(P0.01)。对照组和过表达组LDH活力值分别为108.72±7.20,50.29±10.05,与对照组比较差异有统计学意义(P0.01)。在I/R干预下,凋亡相关蛋白Caspase-3在对照组和过表达组的灰度值分别为99.73±0.61,67.68±0.47,过表达组低于对照组,差异有统计学意义(P0.01)。Bcl-2/Bax在对照组、空载体组和过表达组的灰度值分别为72.52±2.05,39.55±0.46,82.52±1.39,过表达组明显高于对照组和空载体组,差异有统计学意义(P0.01)。在I/R后,自噬相关蛋白Beclin-1在对照组、空载体组和GDF15过表达组的灰度值分别为58.93±0.64,36.71±0.35,88.39±0.72,过表达组明显高于对照组和空载体组,差异有统计学意义(P0.01)。LC3-Ⅱ/LC3-Ⅰ在空载体组和GDF15过表达组的灰度值分别为2.11±0.02,3.32±0.04,过表达组明显高于空载体组,差异有统计学意义(P0.01)。结论上调H9c2细胞的GDF15水平可保护H9c2大鼠心肌细胞,其作用机制可能通过促进细胞的自噬作用得以实现。
[Abstract]:Objective to study the effects of growth differentiation factor 15 (GDF15) on apoptosis and autophagy in H9c2 myocardial ischemia / reperfusion injury by constructing a stable overexpression cardiomyocyte line of GDF15. Methods A model of H9c2I / R injury was established. H9c2 cells were transfected with target GDF15 overexpression lentivirus and negative empty vector virus, and stable overexpression cell lines of GDF15 were selected. The cells were divided into three groups: control group, empty vector group and GDF15 overexpression group. The survival rate of H9c2 cells after I / R was detected by MTT method. The activity of lactate dehydrogenase in the supernatant of cell culture was detected by LDH kit, the expression of apoptosis-related protein Caspase-3Bcl-2nBax and the autophagy associated protein Beclin-1 (LC3-B) were detected by Western blotting. The cell survival rate of empty vector group and overexpression group were 60.47 卤5.03 and 63.21 卤17.13 respectively. The cell activity of overexpressed group was higher than that of control group and empty vector group. The activity of LDH in the control group and overexpression group was 108.72 卤7.20 卤50.29 卤10.05, which was significantly higher than that in the control group. The grayscale value of apoptosis-related protein Caspase-3 in control group and overexpression group was 99.73 卤0.61U 67.68 卤0.47, respectively, which was significantly lower than that in control group (P 0.01). Bcl-2 / Bax was significantly higher in control group. The gray values of empty vector group and overexpression group were 72.52 卤2.05t 39.55 卤0.46U 82.52 卤1.39, respectively, which were significantly higher than those of control group and empty vector group (P 0.01). After I / R, autophagy related protein Beclin-1 was found in control group. The gray values of the empty vector group and the GDF15 overexpression group were 58.93 卤0.64U 36.71 卤0.35 卤88.39 卤0.72, respectively, which were significantly higher than those of the control group and the empty vector group. The grayscale values of P0.01 + LC3- 鈪，

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