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补肾壮督方对大鼠退变髓核组织中miR-155-5p及凋亡相关蛋白的影响
Effects of Bushen Zhuangdufang on the expression of microRNA-155 and mitochondrial apoptotic pathway-related proteins in degenerated nucleus pulposus of rats
投稿时间:2019-05-24  修订日期:2019-06-18
DOI:
中文关键词:  椎间盘退变  髓核细胞  线粒体凋亡通路  补肾壮督方
英文关键词:intervertebral disc degeneration  mitochondrial apoptosis pathway  traditional Chinese medicine  Bushen Zhuangdufang
基金项目:]国家自然科学基金资助项目(81574000)
作者单位E-mail
补肾壮督方对大鼠退变髓核组织中miR--p及凋亡相关蛋白的影响周文明 广州中医药大学 wm13462004@163.com 
林一峰 广州中医药大学附属骨伤科医院 lyf@gzucm.edu.cn 
张震 深圳市沙井人民医院  
白荣飞 广东省中医院  
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中文摘要:
      目的:观察大鼠退变椎间盘髓核组织miR-155-5p及凋亡相关蛋白表达差异,及补肾壮督方对其的影响。方法 SD雄性大鼠,采用纤维环全层针刺造模,随机分为空白对照组,模型组,低剂量中药组、中剂量中药组及高剂量中药组。中药干预4周后,TUNEL法检测退变椎间盘中髓核细胞凋亡率,qRT-PCR检测miR-155-5p及Bax、Bcl-2、Cytc mRNA的表达;Western Blot检测髓核组织中active Caspase-3 ,Bcl-2、Cytc及Bax蛋白表达水平。结果:模型组大鼠髓核细胞凋亡率明显增加(P<0.05),Cytc和Bax mRNA及其蛋白表达量均显著增加(P<0. 05) ,Bcl-2 mRNA及其蛋白表达量显著下降( P<0. 05),Bax/Bcl-2比值显著增加(P<0.05)。中药干预后检测,髓核细胞凋亡率明显减少(P<0.05) ,Cytc和Bax mRNA及其蛋白表达量均明显下降(P<0.05) ,Bcl-2 mRNA及其蛋白显著增加(P<0.05),Bax/Bcl-2比值显著减小(P<0.05)。结论:补肾壮督方可能通过减少active Caspase-3 、Cytc和Bax蛋白表达,增加Bcl-2 蛋白表达,抑制线粒体凋亡通路,从而改善椎间盘退变;miR-155-5p可能参与髓核细胞凋亡的线粒体凋亡通路。
英文摘要:
      Objective:To observe the difference of miR-155-5p and apoptosis-related protein expression in the degenerated intervertebral disc nucleus of rats, and the effect of Bushen Zhuangdufang on it. Methods: SD male rats were randomly divided into blank control group, model group, low-dose Chinese medicine group, medium-dose Chinese medicine group and high-dose Chinese medicine group. Four weeks after the intervention of traditional Chinese medicine, TUNEL method was used to detect the apoptosis rate of nucleus pulposus cells in degenerated intervertebral discs. The expressions of miR-155-5p and Bax, Bcl-2 and Cytc were detected by qRT-PCR. The expression levels of active Caspase-3 , Bcl-2, Cytc and Bax in nucleus pulposus were detected by Western Blot. Results: The apoptosis rate of nucleus pulposus cells in the model group was significantly increased (P<0.05), and the expression of Cytc and Bax mRNA and protein were significantly increased (P<0.05), and the expressions of Bcl-2 mRNA and their proteins were significantly decreased (P<0.05),The ratio of Bax/Bcl-2 increased significantly (P<0.05). After the intervention of traditional Chinese medicine, the apoptosis rate of nucleus pulposus cells was significantly decreased (P<0.05), the expression of Cytc and Bax mRNA and protein were significantly decreased (P<0.05), and the expression of Bcl-2 mRNA and protein was significantly increased. (P<0.05),The ratio of Bax/Bcl-2 decreased significantly (P<0.05). Conclusion: Bushen Zhuangdufang may improve intervertebral disc degeneration by reducing the expression of active Caspase-3 , Cytc and Bax protein, increasing Bcl-2 protein expression and inhibiting mitochondrial apoptosis pathway; miR-155-5p may be involved in nucleus pulposus cell apoptosis mitochondrial apoptosis pathway.
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