心肌细胞氧化损伤对myocardin和核因子E2相关因子2的影响
发布时间：2019-07-31 16:31
【摘要】：利用过氧化氢(H_2O_2)建立大鼠心肌细胞氧化损伤模型;观察心肌细胞氧化损伤过程中myocardin和核因子E2相关因子2(Nrf2)的表达变化并初步探讨myocardin对Nrf2的影响。通过转染质粒过表达目的基因,转染sh RNA质粒下调目的基因表达;通过磺酰罗丹明B(SRB)比色法检测细胞增殖,通过Real-time PCR检测mRNA的表达,通过Western blot检测蛋白的表达。结果显示200μmol/L H_2O_2孵育24 h为最佳H_2O_2氧化损伤条件;H_2O_2抑制myocardin mRNA及蛋白的表达,同时增加Nrf2 mRNA及蛋白的表达;过表达myocardin基因或者下调Nrf2基因后相对活细胞数较对照组明显减少,而下调myocardin基因或者上调Nrf2基因后相对活细胞数较对照组明显增多;过表达myocardin基因后检测到Nrf2 mRNA和蛋白表达出现明显下调,而下调myocardin基因后检测到Nrf2 mRNA和蛋白表达明显上调。因此推断myocardin基因可能抑制细胞增殖,而Nrf2基因可能促进细胞增殖;H_2O_2造成大鼠心肌细胞氧化损伤过程中激活Nrf2相关抗氧化损伤信号途径,其机制可能是通过下调myocardin的表达而实现的。
[Abstract]:The oxidative injury model of cardiomyocytes was established by hydrogen peroxide (H_2O_2), the expression of myocardin and nuclear factor E2 related factor 2 (Nrf2) during oxidative injury of cardiomyocytes was observed, and the effect of myocardin on Nrf2 was discussed. The target gene expression was down-regulated by sh RNA plasmid, cell proliferation was detected by sulfonyl rhodamine B (SRB) colorimetric assay, mRNA expression was detected by Real-time PCR, and protein expression was detected by Western blot. The results showed that 200 渭 mol / L H_2O_2 incubated for 24 h was the best condition for oxidative damage of H_2O_2. H_2O_2 inhibited the expression of myocardin mRNA and protein and increased the expression of Nrf2 mRNA and protein. The number of relative living cells after overexpression of myocardin gene or down-regulation of Nrf2 gene was significantly lower than that of the control group, while the number of relative living cells after down-regulating myocardin gene or up-regulating Nrf2 gene was significantly higher than that of the control group. The expression of Nrf2 mRNA and protein was significantly down-regulated after overexpression of myocardin gene, while the expression of Nrf2 mRNA and protein was up-regulated after down-regulating myocardin gene. Therefore, it is inferred that myocardin gene may inhibit cell proliferation, while Nrf2 gene may promote cell proliferation, and the mechanism of activating Nrf2 related antioxidant damage signal pathway during oxidative damage of rat cardiomyocytes induced by H_2O_2 may be achieved by down-regulating the expression of myocardin.
【作者单位】： 武汉市医疗救治中心结核科;武汉科技大学生物医学研究院;武汉市普仁医院心胸外科;
【基金】：国家自然科学基金资助项目(31471282)
【分类号】：R54
[Abstract]:The oxidative injury model of cardiomyocytes was established by hydrogen peroxide (H_2O_2), the expression of myocardin and nuclear factor E2 related factor 2 (Nrf2) during oxidative injury of cardiomyocytes was observed, and the effect of myocardin on Nrf2 was discussed. The target gene expression was down-regulated by sh RNA plasmid, cell proliferation was detected by sulfonyl rhodamine B (SRB) colorimetric assay, mRNA expression was detected by Real-time PCR, and protein expression was detected by Western blot. The results showed that 200 渭 mol / L H_2O_2 incubated for 24 h was the best condition for oxidative damage of H_2O_2. H_2O_2 inhibited the expression of myocardin mRNA and protein and increased the expression of Nrf2 mRNA and protein. The number of relative living cells after overexpression of myocardin gene or down-regulation of Nrf2 gene was significantly lower than that of the control group, while the number of relative living cells after down-regulating myocardin gene or up-regulating Nrf2 gene was significantly higher than that of the control group. The expression of Nrf2 mRNA and protein was significantly down-regulated after overexpression of myocardin gene, while the expression of Nrf2 mRNA and protein was up-regulated after down-regulating myocardin gene. Therefore, it is inferred that myocardin gene may inhibit cell proliferation, while Nrf2 gene may promote cell proliferation, and the mechanism of activating Nrf2 related antioxidant damage signal pathway during oxidative damage of rat cardiomyocytes induced by H_2O_2 may be achieved by down-regulating the expression of myocardin.
【作者单位】： 武汉市医疗救治中心结核科;武汉科技大学生物医学研究院;武汉市普仁医院心胸外科;
【基金】：国家自然科学基金资助项目(31471282)
【分类号】：R54
【相似文献】
相关期刊论文 前7条
1 牛杰,毛节明,陈明哲,张U，
本文编号：2521446
本文链接：https://www.wllwen.com/yixuelunwen/xxg/2521446.html
最近更新
教材专著
