右美托咪定对人肾小管上皮细胞缺氧复氧时缺氧诱导因子-1α表达的影响  

Effect of dexmedetomidine on expression of hypoxia-inducible factor-1α during hypoxia/reoxygenation in human renal tubular epithelial cells

作　　者：杨春梅[1] 高春霖[1] 于明懂[1] 吕国义[1] 

机构地区：[1]天津医科大学第二医院麻醉科,300211

出　　处：《中华麻醉学杂志》2014年第11期1402-1405,共4页Chinese Journal of Anesthesiology

摘　　要：目的 探讨右美托咪定对人肾小管上皮细胞缺氧复氧时缺氧诱导因子-1α（HIF-1α）表达的影响.方法 体外培养人肾小管上皮细胞（HK-2细胞）,采用随机数字表法将HK-2细胞分为4组（n=24）：正常对照组（C组）：置于37℃常氧恒温细胞培养箱28 h;右美托咪定组（DEX组）：用终浓度0.1 nmol/L的右美托咪定孵育2h,随后置于37℃常氧恒温细胞培养箱28 h;缺氧复氧组（H/R组）：置于37℃厌氧培养罐中缺氧24h后,取出置于37℃常氧恒温细胞培养箱进行复氧4h;缺氧复氧＋右美托咪定组（H/R＋ DEX组）：用终浓度0.1 nmol/L的右美托咪定孵育2h,随后置于37℃厌氧培养罐中缺氧24h后,取出置于37℃常氧恒温细胞培养箱进行复氧4h.各组处理结束后,采用MTT法检测细胞活力,流式细胞仪检测凋亡率,RT-PCR法检测HIF-1α mRNA的表达,Western blot法检测HIF-1α蛋白、活化的caspase-3蛋白的表达水平,并观察细胞生长状况.结果 与C组比较,H/R组和H/R＋DEX组HK-2细胞活力降低,凋亡率升高,HIF-1α mRNA及其蛋白表达上调,活化的caspase-3蛋白表达上调（P＜0.05）,DEX组上述指标差异无统计学意义（P＞0.05）;与H/R组比较,H/R＋ DEX组HK-2细胞活力升高,凋亡率降低,HIF-1α mRNA及其蛋白表达上调,活化的caspase-3蛋白表达下调（P＜0.05）,H/R＋ DEX组细胞状态较H/R组明显改善.结论 右美托咪定减轻人肾小管上皮细胞缺氧复氧损伤的机制可能与上调HIF-1α表达,抑制细胞凋亡有关.Objective To investigate the effect of dexmedetomidine on the expression of hypoxia-inducible factor-1α （HIF-1α） during hypoxia/reoxygenation （H/R） in human renal tubular epithelial cells.Methods Human renal tubular epithelial cells （HK-2 cells） cultured in vitro were randomly divided into 4 groups （n =24 each） using a random number table：control group （group C）,dexmedetomidine group （group DEX）,H/R group and H/R＋ dexmedetomidine group （group H/R ＋ DEX）.In group C,the cells were incubated for 28 h in an incubator filled with normoxia at 37 ℃.In group DEX,dexmedetomidine 0.1 nmol/L （final concentration） was added to the culture medium and the cells were incubated for 2 h,and then incubated for 28 h in an incubator filled with normoxia at 37 ℃.In group H/R,the cells were incubated in an anaerobic chamber for 24 h at 37 ℃,and then incubated for 4 h in an incubator filled with normoxia at 37 ℃.In group H/R ＋ DEX,the cells were incubated for 2 h in the culture medium containing dexmedetomidine 0.1 nmol/L （final concentration）,incubated in an anaerobic chamber for 24 h at 37 ℃,and then incubated for 4 h in an incubator filled with normoxia at 37 ℃.After treatment in each group,the cell viability was measured by MTT assay,cell apoptosis was measured using flow cytometry,the expression of HIF-1α mRNA was detected using RT-PCR,the expression of HIF-1α and activated caspase-3 protein was detected by Western blot,and the cell growth was observed.The apoptosis rate was calculated.Results Compared with group C,the cell viability was significantly decreased,the apoptosis rate was increased,and the expression of HIF-1α mRNA and protein and activated caspase-3 protein was up-regulated in H/.R and H/R ＋ DEX groups,and no significant change was found in group DEX.Compared with group H/R,the cell viability was significantly increased,the apoptosis rate was decreased,the expression of HIF-1α mRNA and protein was up-regulated,the expression of activated caspase-3 protein

关 键 词：右美托咪啶 缺氧诱导因子1 Α亚基 肾小管 上皮细胞 再灌注损伤 

分 类 号：R614[医药卫生—麻醉学]