VDAC2在高位脊髓损伤大鼠心肌细胞线粒体凋亡通路中的作用
发布时间：2018-07-20 12:45
【摘要】：目的观察电压依赖性阴离子通道2(VDAC2)在大鼠高位脊髓损伤后心肌细胞线粒体凋亡通路中的表达,初步探讨VDAC2在高位脊髓损伤后心肌细胞凋亡中的作用。方法健康雄性SD大鼠(清洁级)48只,体重200~250 g。随机分为八组(每组n=6):假手术对照(TC)组:假手术后6 h(TC1)、假手术后12 h(TC2)、假手术后24 h(TC3)和假手术后48 h(TC4);高位脊髓损伤组(TS):SCI损伤后6 h(TS1)、12 h(TS2)、24 h(TS3)和48 h(TS4)。采用改良的Allens打击法,建立高位脊髓损伤大鼠模型,假手术组只暴露脊髓,不进行打击。剪取SD大鼠心尖部心肌组织,使用透射电镜观察心肌细胞的形态学特征;通过原位末端标记法(Tunel法)检测心肌细胞的凋亡率;采用Western blot法检测心肌组织Bax、Bcl-2和VDAC2蛋白的水平,RT-PCR法测定Bax、Bcl-2和VDAC2 m RNA的表达。结果①透射电镜结果显示TC组各时点大部分心肌细胞细胞膜完整;肌原纤维、肌丝排列整齐,清晰可见;线粒体结构完整,未见空泡样变性和肿胀;闰盘结构正常,糖原颗粒丰富,心肌损伤情况明显比TS组同时相轻。②Tunel结果显示TS组较TC组存在明显的细胞凋亡(P0.01)。③Western blot和RT-PCR结果相同,显示TC组Bax、Bcl-2和VDAC2均有表达,各时相点间差异无统计学意义;TS组Bax表达增高,以TS3为著,各时相点与TC组比较差异均有统计学意义(P0.05);TS组Bcl-2表达量于损伤后第6 h、12 h、24 h下降,损伤后第48h表达量恢复,各时相点与TC组比较差异均有统计学意义(P0.05)。同时,TS组Bax/Bcl-2比值增高,TS组与TC组比较差异均有统计学意义(P0.05)。TC组VDAC2有较高水平表达,TS组12 h、24 h、48 h表达受抑制,与TC组比较,差异均有统计学意义(P0.05)。损伤后6 h与同时相TC组比较无显著差异,伤后24 h几乎难以检测到,伤后第48 h开始升高,但仍低于TC组。结论①大鼠高位脊髓损伤后出现心肌细胞凋亡。②大鼠高位脊髓损伤导致的心肌细胞线粒体凋亡通路中,VDAC2和Bcl-2可能起抑制凋亡作用,Bax起促进凋亡作用。
[Abstract]:Objective to investigate the expression of voltage-dependent anion channel 2 (VDAC2) in mitochondria apoptosis pathway after high spinal cord injury (sci) in rats and to explore the role of VDAC2 in myocardial apoptosis after high spinal cord injury (sci). Methods 48 healthy male Sprague-Dawley rats (clean grade), weighing 200 ~ 250g. They were randomly divided into eight groups: sham operation control group (TC): 6 h after sham-operation (TC1), 12 h after sham-operation (TC2), 24 h after sham-operation (TC3) and 48 h after sham-operation (TC4), and 6 h (TS1) 12 h (TS2) 24 h (TS3) and 48 h (TS4) after high spinal cord injury (TS). The rat model of high spinal cord injury was established by modified Allens method. The sham operation group only exposed the spinal cord and did not attack. The apical myocardial tissue of SD rats was cut and the morphologic characteristics of cardiomyocytes were observed by transmission electron microscope (TEM), and the apoptosis rate of cardiomyocytes was detected by Tunel method. The expression of BaxanBcl-2 and VDAC2 mRNA in myocardium was detected by Western blot and RT-PCR. Results 1 the results of transmission electron microscope showed that most of the myocardial cell membrane was intact at each time point in TC group; the myofibrils, myofilaments were arranged neatly and clearly; the mitochondria structure was intact without vacuolar degeneration and swelling; the intercalated disc structure was normal, and the structure of intercalated disc was normal. Results of myocardial injury in TS group were significantly lighter than those in TS group. The results of apoptosis (P0.01) .3Western blot and RT-PCR showed that the expression of BaxanBcl-2 and VDAC2 in TC group was the same as that in TS group. The expression of Bax in TS group was significantly higher than that in TC group (P0.05). The expression of Bcl-2 in TS group decreased at 6 h and 12 h after injury, and recovered at 48 h after injury. There was significant difference between each time point and TC group (P0.05). At the same time, the increase of Bax-Bcl-2 ratio in TS group and TC group were statistically significant (P0.05). There was a high level of VDAC2 expression in TS group. The expression of VDAC2 in TS group was inhibited for 24 h or 48 h, compared with TC group, the difference was statistically significant (P0.05). There was no significant difference between the six hours after injury and that in the simultaneous phase TC group. It was almost difficult to detect at 24 hours after injury, but it began to rise at 48 hours after injury, but still lower than that in the TC group. Conclusion 1 apoptosis of cardiomyocytes induced by high spinal cord injury in rats. 2 in mitochondria apoptosis pathway induced by high spinal cord injury in rats, VDAC2 and Bcl-2 may inhibit apoptosis and Bax may promote apoptosis.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R651.2
本文编号：2133555
[Abstract]:Objective to investigate the expression of voltage-dependent anion channel 2 (VDAC2) in mitochondria apoptosis pathway after high spinal cord injury (sci) in rats and to explore the role of VDAC2 in myocardial apoptosis after high spinal cord injury (sci). Methods 48 healthy male Sprague-Dawley rats (clean grade), weighing 200 ~ 250g. They were randomly divided into eight groups: sham operation control group (TC): 6 h after sham-operation (TC1), 12 h after sham-operation (TC2), 24 h after sham-operation (TC3) and 48 h after sham-operation (TC4), and 6 h (TS1) 12 h (TS2) 24 h (TS3) and 48 h (TS4) after high spinal cord injury (TS). The rat model of high spinal cord injury was established by modified Allens method. The sham operation group only exposed the spinal cord and did not attack. The apical myocardial tissue of SD rats was cut and the morphologic characteristics of cardiomyocytes were observed by transmission electron microscope (TEM), and the apoptosis rate of cardiomyocytes was detected by Tunel method. The expression of BaxanBcl-2 and VDAC2 mRNA in myocardium was detected by Western blot and RT-PCR. Results 1 the results of transmission electron microscope showed that most of the myocardial cell membrane was intact at each time point in TC group; the myofibrils, myofilaments were arranged neatly and clearly; the mitochondria structure was intact without vacuolar degeneration and swelling; the intercalated disc structure was normal, and the structure of intercalated disc was normal. Results of myocardial injury in TS group were significantly lighter than those in TS group. The results of apoptosis (P0.01) .3Western blot and RT-PCR showed that the expression of BaxanBcl-2 and VDAC2 in TC group was the same as that in TS group. The expression of Bax in TS group was significantly higher than that in TC group (P0.05). The expression of Bcl-2 in TS group decreased at 6 h and 12 h after injury, and recovered at 48 h after injury. There was significant difference between each time point and TC group (P0.05). At the same time, the increase of Bax-Bcl-2 ratio in TS group and TC group were statistically significant (P0.05). There was a high level of VDAC2 expression in TS group. The expression of VDAC2 in TS group was inhibited for 24 h or 48 h, compared with TC group, the difference was statistically significant (P0.05). There was no significant difference between the six hours after injury and that in the simultaneous phase TC group. It was almost difficult to detect at 24 hours after injury, but it began to rise at 48 hours after injury, but still lower than that in the TC group. Conclusion 1 apoptosis of cardiomyocytes induced by high spinal cord injury in rats. 2 in mitochondria apoptosis pathway induced by high spinal cord injury in rats, VDAC2 and Bcl-2 may inhibit apoptosis and Bax may promote apoptosis.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R651.2
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