瑞芬太尼后处理对糖尿病性心肌细胞保护作用削弱的机制：与组蛋白去乙酰化酶3表达的关系  

Mechanism underlying mitigation of remifentanil postconditioning-induced protection of diabetic cardiomyocytes： the relationship with histone deacetylase 3 expression

作　　者：刘芹[1] 陈满丽[1] 顾尔伟[1] 陈立建[1] 张雷[1] 都建[1] 程新琦[1] 

机构地区：[1]安徽医科大学第一附属医院麻醉科,合肥市230022

出　　处：《中华麻醉学杂志》2016年第7期851-854,共4页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金（81341014）

摘　　要：目的评价组蛋白去乙酰化酶3（HDAC3）与瑞芬太尼后处理对糖尿病性心肌细胞保护作用削弱机制的关系。方法H9c2细胞在10％胎牛血清DMEM／F12培养基中培养、传代，接种于6孔板（2ml／孔），接种密度10^5个／ml，接种12h后细胞贴壁，换正常糖（5．5mmol／L）或高糖（25mmol／L）DMEM培养基培养48h。采用随机数字表法分为6组（n=18）：对照组（CON组）、缺氧复氧组（H／R组）、瑞芬太尼后处理组（RPC组）、高糖组（HG组）、高糖＋缺氧复氧组（HG—H／R组）和高糖＋瑞芬太尼后处理组（HG—RPC组）。H／R组、RPC组、HG—H／R组和HG—RPC组弃去培养液，加入Ty-rode液，将培养板置于95％N2-5％CO2培养罐中孵育5h，H／R组和HG—H／R组更换为含10％胎牛血清的相应糖DMEM培养基孵育1h，RPC组和HG—RPC组更换为含终浓度为1μmol／L瑞芬太尼的DMEM培养基孵育1h。于复氧1h时，采用CCK-8法检测细胞活力，Annexin V．FITC／PI流式细胞术检测细胞凋亡率，Western blot法检测细胞HDAC3和caspase-3表达水平。结果与CON组比较，H／R组细胞活力降低，细胞凋亡率增加，easpase-3与HDAC3表达上调（P〈0．05）；与H／R组比较，RPC组细胞活力升高，细胞凋亡率下降，easpase-3和HDAC3表达下调（P〈0．05）；与HG组比较，HG—H／R组细胞活力降低，细胞凋亡率升高，easpase-3和HDAC3表达上调（P〈0．05）；与HG—H／R组比较，HG-RPC组细胞活力、细胞凋亡率、caspase-3与HDAC3表达差异无统计学意义（P〉0．05）。结论瑞芬太尼后处理对糖尿病性心肌细胞保护作用削弱的机制与高糖上调HDAC3表达有关。Objective To evaluate the relationship between histone deacetylase 3 （HDAC3） expression and the mechanism underlying mitigation of remifentanil postconditioning-induced protection of diabetic cardiomyocytes. Methods H9c2 cells were cultured in DMEM/F12 culture medium supplemented with 10% fetal bovine serum. The ceils were seeded in 6-well plates （2 ml/well） at a density of 105 cells/ml. After the cells were cultured for 12 h, the cells were attached to the wall and cultured for 48 h in the normoglycemie （5.5 mmol/L） or hyperglycemic （25 mmol/L） DMEM culture medium. The ceils were then randomly divided into 6 groups （n = 18 each） using a random number table： control group （group CON） , hypoxia/reoxygenation group （ group H/R） , remifentanil posteonditioning group （ group RPC） , hyperglycemia group （group HG）, hyperglycemia p/us hypoxia/reoxygenation group （group HG-H/R）, and hyperglycemia plus remifentanil postconditioning group （group HG-RPC）. In H/R, RPC, HG-H/R and HG-RPC groups, the cells were exposed to 95% N2-5% CO2 in an incubator for 5 h after changing the culture medium for Tyrode solution. In H/R and HG-H/R groups, the culture medium was changed to the DMEM/F12 culture medium supplemented with 10% fetal bovine serum and glucose at the corresponding concentration, and the cells were then incubated for 1 h. In RPC and HG-RPC groups, the cells were incubated in the DMEM culture medium containing remifentanil at the final concentration of 1 μmol/L, and the cells were then incubated for 1 h. At 1 h of reoxygenation, the cell viability was measured by CCK-8 assay, the cell apoptosis was detected by AnnexinV-FITC/PI flow eytometry, and the expression of HDAC3 and caspase-3 in cells was detected, by Western blot. The apoptotie rate was calculated. Results Compared with group CON, the cell viability was significantly decreased, the cell apoptotic rate was significantly increased, and the expression of caspase-3 and HDAC3 was significantly up-regulated in group H/R �

关 键 词：哌啶类 糖尿病 肌细胞 心脏 缺氧 组蛋白脱乙酰基酶类 缺血后处理 

分 类 号：R614[医药卫生—麻醉学]