低浓度过氧化氢对心肌细胞自噬的影响及其机理研究

发布时间：2018-12-11 02:19

【摘要】：目的:1、低浓度过氧化氢对心肌细胞自噬的影响。2、低浓度过氧化氢对心肌细胞自噬相关蛋白Beclin1的影响。3、抗氧化剂对压力超负荷早期大鼠心脏心肌细胞自噬的影响。方法:分组:1、H9C2心肌细胞培养模型:(1)正常对照组(2)10u M过氧化氢干预组2、大鼠压力超负荷模型:(1)假手术组(sham)(2)腹主动脉缩窄(AAC)组(3)AAC+N-乙酰半胱胺酸(NAC)组采用Western Blot蛋白印迹法检测心肌细胞蛋白量的表达。结果:离体实验:H9C2心肌细胞培养模型中,低浓度H2O2(10umol/L)干预,72h对照组(1.00±0.10)VS 10umol/L组(0.76±0.12),组间比较,P0.05,显示LC3II/I值在10umol/L H2O2干预72h时与对照组相比,表达明显下降;低浓度H2O2(10umol/L)干预,48h对照组(1.00±0.11)VS 10umol/L组(0.76±0.08),组间比较,P0.05,显示Beclin1在10umol/L H2O2干预48h时与对照组相比,表达明显下降。在体实验:大鼠压力超负荷模型中,AAC术后3天,全心重量和左心室重量有增加的趋势,抗氧化剂NAC减轻心脏重量的增加。蛋白免疫印迹方法中,sham组(1.00±0.10)VS AAC组(0.61±0.12),组间比较,P0.05,显示LC3II/I在AAC组与sham组比较,表达明显下降,具有统计学意义;AAC组(0.61±0.12)VS AAC+NAC组(1.01±0.10),组间比较,P0.05,显示LC3II/I在AAC+NAC组与AAC组比较,表达明显上升,具有统计学意义。sham组(1.00±0.08)VS AAC组(0.81±0.06),组间比较,P0.05,显示Beclin1在AAC组与sham组比较,表达显著降低,具有统计学意义;AAC组(0.81±0.06)VS AAC+NAC组(1.01±0.08),组间比较,P0.05,显示Beclin1在AAC+NAC组与AAC组比较,表达明显升高,具有统计学意义。结论:1、低浓度过氧化氢抑制心肌细胞自噬的发生。2、在压力超负荷早期,大鼠心肌细胞自噬降低。3、抗氧化剂可抑制压力超负荷早期所致心肌细胞自噬的降低。
[Abstract]:Objective: 1, the effect of low concentration hydrogen peroxide on cardiac myocyte autophagy. 2. The effect of low concentration hydrogen peroxide on cardiac myocyte autophagy protein (Beclin1). 3. The effect of antioxidant on cardiac myocyte autophagy in rats at the early stage of pressure overload. Methods: the model of H9C2 cardiomyocyte culture was divided into two groups: (1) normal control group (2) 10u M hydrogen peroxide treatment group (2), Rat pressure overload model: (1) (sham) (_ 2) abdominal aorta constricted (AAC) group (3) AAC N-acetylcysteine (NAC) group was used to detect the expression of cardiomyocyte protein by Western Blot Western blot. Results: in vitro experiment: in H9C2 cardiomyocyte culture model, low concentration H2O2 (10umol/L) intervention, 72 h control group (1.00 卤0.10) VS 10umol/L group (0.76 卤0.12). The results showed that the expression of LC3II/I was significantly lower than that of the control group at 72 h after 10umol/L H2O2 intervention. Low concentration H2O2 (10umol/L) intervention, 48 h control group (1.00 卤0.11) VS 10umol/L group (0.76 卤0.08), the comparison between groups, P0.05, which showed that the expression of Beclin1 decreased significantly after 48 h of 10umol/L H2O2 intervention compared with the control group. In vivo experiment, the whole heart weight and left ventricular weight increased 3 days after AAC in rat model of pressure overload. The antioxidant NAC alleviated the increase of cardiac weight. In Western blot, the expression of LC3II/I in sham group (1.00 卤0.10) VS AAC group, 0.61 卤0.12) was significantly lower than that in AAC group (0.61 卤0.12), which showed that the expression of LC3II/I in AAC group was significantly lower than that in sham group (P 0.05). The expression of LC3II/I in AAC group (0.61 卤0.12) VS AAC NAC group, 1.01 卤0.10) was significantly higher than that in AAC NAC group and AAC group (P 0.05). The expression of Beclin1 in sham group (1.00 卤0.08) VS AAC group, P 0.05) was significantly lower than that in AAC group (0.81 卤0.06), which was statistically significant. The expression of Beclin1 in AAC group (0.81 卤0.06) VS AAC NAC group, 1.01 卤0.08) was significantly higher than that in AAC NAC group (1.01 卤0.08), which showed that the expression of Beclin1 in AAC NAC group was significantly higher than that in AAC group (P 0.05). Conclusion: 1. Low concentration of hydrogen peroxide inhibits the occurrence of cardiac myocyte autophagy. 2. At the early stage of pressure overload, cardiac myocyte autophagy decreases in rats. 3. Antioxidant can inhibit the decrease of cardiac myocyte autophagy induced by pressure overload.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R54

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