VDAC2在高位脊髓損傷大鼠心肌細(xì)胞線粒體凋亡通路中的作用
發(fā)布時(shí)間：2018-07-20 12:45
【摘要】：目的觀察電壓依賴性陰離子通道2(VDAC2)在大鼠高位脊髓損傷后心肌細(xì)胞線粒體凋亡通路中的表達(dá),初步探討VDAC2在高位脊髓損傷后心肌細(xì)胞凋亡中的作用。方法健康雄性SD大鼠(清潔級(jí))48只,體重200~250 g。隨機(jī)分為八組(每組n=6):假手術(shù)對(duì)照(TC)組:假手術(shù)后6 h(TC1)、假手術(shù)后12 h(TC2)、假手術(shù)后24 h(TC3)和假手術(shù)后48 h(TC4);高位脊髓損傷組(TS):SCI損傷后6 h(TS1)、12 h(TS2)、24 h(TS3)和48 h(TS4)。采用改良的Allens打擊法,建立高位脊髓損傷大鼠模型,假手術(shù)組只暴露脊髓,不進(jìn)行打擊。剪取SD大鼠心尖部心肌組織,使用透射電鏡觀察心肌細(xì)胞的形態(tài)學(xué)特征;通過(guò)原位末端標(biāo)記法(Tunel法)檢測(cè)心肌細(xì)胞的凋亡率;采用Western blot法檢測(cè)心肌組織Bax、Bcl-2和VDAC2蛋白的水平,RT-PCR法測(cè)定Bax、Bcl-2和VDAC2 m RNA的表達(dá)。結(jié)果①透射電鏡結(jié)果顯示TC組各時(shí)點(diǎn)大部分心肌細(xì)胞細(xì)胞膜完整;肌原纖維、肌絲排列整齊,清晰可見(jiàn);線粒體結(jié)構(gòu)完整,未見(jiàn)空泡樣變性和腫脹;閏盤結(jié)構(gòu)正常,糖原顆粒豐富,心肌損傷情況明顯比TS組同時(shí)相輕。②Tunel結(jié)果顯示TS組較TC組存在明顯的細(xì)胞凋亡(P0.01)。③Western blot和RT-PCR結(jié)果相同,顯示TC組Bax、Bcl-2和VDAC2均有表達(dá),各時(shí)相點(diǎn)間差異無(wú)統(tǒng)計(jì)學(xué)意義;TS組Bax表達(dá)增高,以TS3為著,各時(shí)相點(diǎn)與TC組比較差異均有統(tǒng)計(jì)學(xué)意義(P0.05);TS組Bcl-2表達(dá)量于損傷后第6 h、12 h、24 h下降,損傷后第48h表達(dá)量恢復(fù),各時(shí)相點(diǎn)與TC組比較差異均有統(tǒng)計(jì)學(xué)意義(P0.05)。同時(shí),TS組Bax/Bcl-2比值增高,TS組與TC組比較差異均有統(tǒng)計(jì)學(xué)意義(P0.05)。TC組VDAC2有較高水平表達(dá),TS組12 h、24 h、48 h表達(dá)受抑制,與TC組比較,差異均有統(tǒng)計(jì)學(xué)意義(P0.05)。損傷后6 h與同時(shí)相TC組比較無(wú)顯著差異,傷后24 h幾乎難以檢測(cè)到,傷后第48 h開(kāi)始升高,但仍低于TC組。結(jié)論①大鼠高位脊髓損傷后出現(xiàn)心肌細(xì)胞凋亡。②大鼠高位脊髓損傷導(dǎo)致的心肌細(xì)胞線粒體凋亡通路中,VDAC2和Bcl-2可能起抑制凋亡作用,Bax起促進(jìn)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.
【學(xué)位授予單位】：福建醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R651.2
本文編號(hào)：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.
【學(xué)位授予單位】：福建醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R651.2
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