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-  2018 

组蛋白去乙酰化酶6抑制剂对糖尿病性心肌细胞缺氧复氧损伤的保护作用及其机制研究 Inhibition of Histone Deacetylase 6 Activity Confers Protective Effects Against Hypoxia-Reoxygenation Injury in Diabetic Myocardiocytes

Keywords: 组蛋白去乙酰化酶6,蛋白激酶B,糖尿病,心肌缺血再灌注

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Abstract:

目的:评价组蛋白去乙酰化酶6(HDAC6)抑制剂Tubacin(TBA)对糖尿病性心肌细胞缺氧复氧损伤的保护作用和机制。方法:正常培养的H9c2细胞,采用随机数字表法分为4组(n=24):高糖常氧组(N组)、高糖缺氧复氧组(H/R组)、高糖缺氧复氧+Tubacin组(H/R+TBA组)、高糖缺氧复氧+Tubacin+Akt抑制剂MK-2206(H/R+TBA+MK组)。高糖培养基(葡萄糖浓度33mmol/L)培养H9c2细胞24h。H9c2心肌细胞缺氧(95%N2+5%CO_2)4h,复氧(90%O2+10%CO_2)2h制备H9c2心肌细胞缺氧复氧损伤模型。TBA组H/R前6h给予Tubacin(2μmol/L)。Akt抑制剂组H/R前1h给予MK-2206(300nmol/L)。采用ELISA法检测培养上清液乳酸脱氢酶(LDH)含量,采用CCK-8试剂盒检测细胞存活率,HDAC6活性检测试剂盒检测心肌细胞HDAC6活性。Western Blot法检测磷酸化蛋白激酶B (p-Akt,ser473)、总蛋白激酶B(t-Akt)、磷酸化叉头蛋白O3a(p-Foxo3a,ser253)和总叉头蛋白O3a(t-Foxo3a)的表达水平。结果:与N组比较,H/R组LDH释放量和细胞凋亡水平增加(P<0.05),p-Akt、t-Akt、p-Foxo3a、t-Foxo3a蛋白表达无显著差异(P>0.05)。与H/R组比较,H/R+TBA组LDH释放量和细胞凋亡水平降低(P<0.05),同时p-Akt和p-Foxo3a表达升高,t-Akt和t-Foxo3a无显著差异(P>0.05)。与H/R+TBA组相比,H/R+TBA+MK组LDH释放量和细胞凋亡水平升高(P<0.05),同时p-Akt和p-Foxo3a表达显著降低(P<0.05),t-Akt和t-Foxo3a无显著差异(P>0.05)。结论:HDAC6抑制剂TBA对糖尿病性心肌细胞缺氧复氧损伤的保护效应是通过激活Akt/Foxo3a通路而产生的

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