七氟醚预处理对H9C2心肌细胞缺氧/复氧后转录沉默信息调节器3的表达及乙酰化水平的影响

doi:10.6040/j.issn.1671-7554.0.2018.877

摘要/Abstract

摘要： 目的探究七氟醚预处理对大鼠H9C2心肌细胞缺氧/复氧后,对转录沉默信息调节器3(SIRT3)的表达及细胞蛋白乙酰化的影响。方法将大鼠H9C2心肌细胞随机分为对照组、缺氧/复氧组和七氟醚预处理组。低氧培养箱构建缺氧/复氧模型:对照组心肌细胞无任何处理;缺氧/复氧组心肌细胞缺氧2 h/复氧2 h;七氟醚预处理组心肌细胞在缺氧/复氧前予以2.5%七氟醚预处理1 h。四甲基偶氮唑蓝(MTT)检测各组细胞存活率;采用JC-1荧光探针检测线粒体膜电位;采用Western blotting法检测细胞整体蛋白及线粒体蛋白乙酰化水平、SIRT3表达及线粒体自噬水平的变化。结果缺氧/复氧组较对照组心肌细胞SIRT3(0.78±0.04 vs 1.04±0.06)表达减少,细胞整体蛋白(1.72±0.06 vs 0.98±0.03 )及线粒体蛋白(0.96±0.03 vs 0.45±0.03)乙酰化水平升高(P<0.05);同时心肌细胞存活率［(48.2±0.4)% vs 100%］、线粒体膜电位(1.72±0.14 vs 2.83±0.11)下降,线粒体自噬水平升高(P<0.05);与缺氧/复氧组相比,七氟醚预处理组心肌细胞SIRT3表达增加(0.93±0.03 vs 0.78±0.04),细胞整体蛋白(1.34±0.05 vs 1.72±0.06)及线粒体蛋白(0.65±0.04 vs 0.96±0.03)乙酰化水平降低(P<0.05);同时心肌细胞存活率［(65.80±1.53)% vs(48.20±0.40)%］及线粒体膜电位(2.33±0.12 vs 1.72±0.14)升高,线粒体自噬激活水平降低(P<0.05)。结论七氟醚预处理可以增加SIRT3表达,降低心肌细胞缺氧/复氧损伤后蛋白的乙酰化水平,可能是七氟醚预处理对心肌细胞保护作用的潜在分子机制。

关键词: 七氟醚预处理, 乙酰化, 转录沉默信息调节器3, 线粒体自噬, 缺氧/复氧

Abstract: Objective To investigate the effects of sevoflurane preconditioning on silent information regulator of transcription 3(SIRT3)expression and the acetylation of H9C2 cardiomyocytes after hypoxia/reoxygenation. Methods Rat H9C2 cardiomyocytes were randomly divided into control group, hypoxia/reoxygenation group and sevoflurane pretreatment group. Cells without any treatment served as control group. Hypoxia/reoxygenation models were prepared in a hypoxia incubator, in which cells were exposed to 2 h hypoxia followed by 2 h reoxygenation. Cells in the sevoflurane pretreatment group were treated with 2.5% sevoflurane for 1 h before hypoxia/reoxygenation processing. The cell survival 山东大学学报 (医学版)57卷3期 -熊超,等.七氟醚预处理对H9C2心肌细胞缺氧/复氧后转录沉默信息调节器3的表达及乙酰化水平的影响 \=-was detected with methylthiazolyltetrazolium(MTT)assay. Mitochondrial membrane potential was examined with JC-1 fluorescence probe. The acetylation of proteins, especially that of mitochondrial protein, SIRT3 expression, and mitophagy level were detected with Western blotting. Results Compared with the control group, the hypoxia/reoxygenation group had decreased expression of SIRT3(0.78±0.04 vs 1.04±0.06), increased acetylation level of proteins(1.72±0.06 vs 0.98±0.03)and mitochondrial protein(0.96±0.03 vs 0.45±0.03)(P<0.05), decreased cell survival ［(48.2±0.4)% vs 100%］, decreased mitochondrial membrane potential(1.72±0.14 vs 2.83±0.11), and increased mitophagy level(P<0.05). Compared with the hypoxia/reoxygenation group, the sevoflurane pretreatment group had increased expression of SIRT3(0.93±0.03 vs 0.78±0.04), decreased acetylation level of proteins(1.34±0.05 vs 1.72±0.06)and mitochondrial protein(0.65±0.04 vs 0.96±0.03)(P<0.05), increased cell survival ［(65.80±1.53)% vs (48.20±0.40)%］ and mitochondrial membrane potential(2.33±0.12 vs 1.72±0.14), and decreased mitophagy level(P<0.05). Conclusion Sevoflurane pretreatment is capable of elevating the expression of SIRT3 and downregulating the acetylation of H9C2 cells after hypoxia/reoxygenation injury, which might be the molecular mechanism underlying the protective effects of sevoflurane on cardiomyocytes.

Key words: Sevoflurane preconditioning, Acetylization, Silent information regulator of transcription 3, Mitophagy, Hypoxia/reoxygenation

中图分类号:

R197.1

熊超,刘力,冯建国,魏继承. 七氟醚预处理对H9C2心肌细胞缺氧/复氧后转录沉默信息调节器3的表达及乙酰化水平的影响[J]. 山东大学学报 (医学版), 2019, 57(3): 25-30.

XIONG Chao, LIU Li, FENG Jianguo, WEI Jicheng. Effects of sevoflurane preconditioning on silent information regulator of transcription 3 expression and acetylation in H9C2 cardiomyocytes after hypoxia/reoxygenation[J]. Journal of Shandong University (Health Sciences), 2019, 57(3): 25-30.

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