【摘要】：利用過氧化氫(H_2O_2)建立大鼠心肌細(xì)胞氧化損傷模型;觀察心肌細(xì)胞氧化損傷過程中myocardin和核因子E2相關(guān)因子2(Nrf2)的表達(dá)變化并初步探討myocardin對(duì)Nrf2的影響。通過轉(zhuǎn)染質(zhì)粒過表達(dá)目的基因,轉(zhuǎn)染sh RNA質(zhì)粒下調(diào)目的基因表達(dá);通過磺酰羅丹明B(SRB)比色法檢測(cè)細(xì)胞增殖,通過Real-time PCR檢測(cè)mRNA的表達(dá),通過Western blot檢測(cè)蛋白的表達(dá)。結(jié)果顯示200μmol/L H_2O_2孵育24 h為最佳H_2O_2氧化損傷條件;H_2O_2抑制myocardin mRNA及蛋白的表達(dá),同時(shí)增加Nrf2 mRNA及蛋白的表達(dá);過表達(dá)myocardin基因或者下調(diào)Nrf2基因后相對(duì)活細(xì)胞數(shù)較對(duì)照組明顯減少,而下調(diào)myocardin基因或者上調(diào)Nrf2基因后相對(duì)活細(xì)胞數(shù)較對(duì)照組明顯增多;過表達(dá)myocardin基因后檢測(cè)到Nrf2 mRNA和蛋白表達(dá)出現(xiàn)明顯下調(diào),而下調(diào)myocardin基因后檢測(cè)到Nrf2 mRNA和蛋白表達(dá)明顯上調(diào)。因此推斷myocardin基因可能抑制細(xì)胞增殖,而Nrf2基因可能促進(jìn)細(xì)胞增殖;H_2O_2造成大鼠心肌細(xì)胞氧化損傷過程中激活Nrf2相關(guān)抗氧化損傷信號(hào)途徑,其機(jī)制可能是通過下調(diào)myocardin的表達(dá)而實(shí)現(xiàn)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.
【作者單位】： 武漢市醫(yī)療救治中心結(jié)核科;武漢科技大學(xué)生物醫(yī)學(xué)研究院;武漢市普仁醫(yī)院心胸外科;
【基金】：國家自然科學(xué)基金資助項(xiàng)目(31471282)
【分類號(hào)】：R54

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