刺囊酸对缺氧复氧诱导心肌细胞凋亡相关基因表达的影响
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广东省自然科学基金(8451008901000788)


Effects of Echinocystic Acid on the Expression of Apoptosis-Associated Gene in the Rat Cardiomyocytes Subjected to Anoxia /Reoxygenation Injury
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    摘要:

    目的 研究刺囊酸预处理对原代培养的SD乳鼠心肌细胞缺氧复氧损伤诱导的心肌细胞凋亡相关基因Bcl-2、Bax及聚二磷酸腺苷核糖聚合酶(PARP 89 kDa)表达的影响。方法 实验随机分为6组:对照组,缺氧复氧损伤组,缺氧预处理组及低、中、高剂量刺囊酸组(0.5 μmol/L、5 μmol/L和50 μmol/L),分别予以缺氧3 h后再复氧2 h,检测细胞存活率、乳酸脱氢酶(LDH)活性,Western Blot杂交法检测心肌细胞Bcl-2、Bax及PARP(89 kDa)蛋白表达变化。结果 与对照组比较,缺氧复氧损伤组心肌细胞Bcl-2蛋白表达显著低于对照组(P<0.01),Bax 蛋白及PARP(89 kDa)表达显著高于对照组( P<0.01),细胞存活率明显低于对照组( P<0.05)。与缺氧复氧损伤组比较,不同剂量的刺囊酸预处理能显著提高细胞存活率,降低LDH活性,呈剂量依赖性;中剂量刺囊酸显著增加Bcl-2蛋白表达 (P<0.05),抑制Bax及PARP(89 kDa)蛋白表达(P<0.05)。结论 刺囊酸预处理可通过上调Bcl-2蛋白表达,抑制PARP(89 kDa)及Bax蛋白表达,抑制细胞凋亡,对抗心肌细胞缺氧复氧损伤。

    Abstract:

    Aim To study the effects of Echinocystic acid on the protein expression of apoptosis-associated Bcl-2,Bax and PARP (89 kDa) in the primary cultured rat cardiomyocytes subjected to anoxia/reoxygenation injury. Methods The cardiomyocytes were divided into normal group,anoxia/reoxygenation injury group,anoxia/reoxygenation preconditioning group,different concentration Echinocystic acid groups (0.5 μmol/L,5 μmol/L and 50 μmol/L). Cardiomyocytes were subjected to anoxia for 3 hours and subsequently reoxygenation for 2 hours. Cell viability and LDH activity in medium were measured. The expression o f Bcl-2 and the apoptotic protein Bax and PARP (89 kDa) were detected by Western Blot. Results Compared with that of the control group,the expression of Bcl-2 protein in cardiomyocytes decreased significantly (P<0.01) and the expression of Bax protein and PARP (89 kDa) in cardiomyocytes increased significantly (P<0.01) after reoxygenation. Cell viability decreased obviously (P<0.05). Compared with that of the anoxia/reoxygenation group,pretreatment with different concentration Echinocystic acid decreased LDH activity and increased cell viability(P<0.05),the expression of Bcl-2 protein in the Echinocystic acid (5 μmol/L) groups increased significantly (P<0.05) and the expression of Bax protein decreased significantly (P<0.05). Conclusion The antiapopotic effects of Echinocystic acid might be attributed to the up-regulated expression of Bcl-2 gene and the inhibited expression of Bax and PARP(89 kDa) gene expression.

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陈 杰,沈映冰,许 静,官碧琼,罗子玲.刺囊酸对缺氧复氧诱导心肌细胞凋亡相关基因表达的影响[J].中国动脉硬化杂志,2013,21(02):135~138.

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  • 收稿日期:2012-01-26
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