高糖H9c2运用七氟醚后处理时HIF-1α对mPTP通道调节机制的研究

发布时间：2018-03-10 07:13

本文选题：高糖状态心肌细胞　切入点：七氟醚后处理　出处：《新疆医科大学》2016年硕士论文　论文类型：学位论文

【摘要】：目的:探讨高糖状态心肌细胞低氧诱导因子1α(HIF-1α)对线粒体膜通透性转化孔(mPTP)的调节机制,并揭示高糖状态下心肌缺血再灌注损伤加重的关键分子靶点。方法:运用H9c2心肌细胞系建立缺氧复氧损伤模型,采用试剂盒检测乳酸脱氢酶(LDH)及CCK-8水平;TUNEL法检测细胞凋亡;通过激光共聚焦检测细胞线粒体膜通透性转化孔及线粒体膜电位改变;通过Western blot法检测HIF-1α及己糖激酶(HKII)的表达;免疫荧光法检测HKII含量及其分布。结果:1)与正常组相比较,缺氧复氧损伤及七氟醚后处理均可上调HIF-1α蛋白表达,其中七氟醚上调更为显著,HIF-1α可抑制mPTP开放,减少细胞凋亡;2)抑制HIF-1α其下游靶基因HKII表达受到明显抑制,mPTP通道开放增加,细胞损伤明显;3)抑制HKII后,HIF-1α丧失对mPTP的调节,七氟醚后处理失去其心肌保护作用;4)在高糖状态下,运用七氟醚后处理,并未减少心肌细胞凋亡。其HIF-1α及HKII的表达较正常七氟醚处理组无明显差异,但HKII存在向细胞膜聚集现象。结论:在缺氧复氧损伤中七氟醚后处理可上调HIF-1α蛋白表达,产生心肌保护作用。其机制可能是上调HIF-1α能够激活其下游靶基因HKII,抑制mPTP通道的开放,从而减轻缺氧复氧造成的心肌细胞损伤。高糖状态下七氟醚保护作用减弱或消失的原因可能是由于HKII发生转位导致HIF-1α失去了对mPTP通道的调控作用。从而增加了高糖状态下缺氧复氧损伤心肌细胞的坏死与凋亡。
[Abstract]:Aim: to investigate the regulatory mechanism of hypoxia inducible factor-1 伪 (HIF-1 伪) on mitochondrial membrane permeability transforming pore (mPTP) in hyperglycemic cardiomyocytes. Methods: the model of hypoxic reoxygenation injury was established by H9c2 myocardial cell line, the lactate dehydrogenase (LDHs) and CCK-8 level were detected by Tunel method. The changes of mitochondrial membrane permeability transformation pore and mitochondrial membrane potential were detected by laser confocal focusing, the expression of HIF-1 伪 and hexokinase II was detected by Western blot method, and the content and distribution of HKII were detected by immunofluorescence. Hypoxia reoxygenation injury and sevoflurane post-treatment could up-regulate the expression of HIF-1 伪 protein, especially sevoflurane up-regulated HIF-1 伪 inhibited the opening of mPTP and decreased apoptosis of HIF-1 伪. After treatment with sevoflurane, the protective effect of sevoflurane on mPTP was lost. In high glucose state, sevoflurane was treated with sevoflurane. The expression of HIF-1 伪 and HKII was not significantly different from that of the normal sevoflurane treatment group, but the expression of HKII accumulated to the cell membrane. Conclusion: sevoflurane post-treatment can up-regulate the expression of HIF-1 伪 protein during hypoxia reoxygenation injury. The mechanism may be that up-regulation of HIF-1 伪 can activate its downstream target gene HKII and inhibit the opening of mPTP channel. The protective effect of sevoflurane on cardiomyocytes induced by hypoxia and reoxygenation could be reduced or disappeared due to the transposition of HKII, which resulted in the loss of regulation of HIF-1 伪 on mPTP channels. Necrosis and apoptosis of cardiomyocytes injured by hypoxia and reoxygenation in glucose state.
【学位授予单位】：新疆医科大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R614

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