中文摘要：

目的探讨高血糖对人肾小管上皮细胞活性氧（ROS）产生及凋亡的影响。方法将人近端肾小管上皮细胞（HKC）分别以不同时间在含有5.5 mM D（右旋）-葡萄糖（正常糖对照组）、25 mMD-葡萄糖（高糖组）和20 mML（左旋）-葡萄糖＋5.5 mMD-葡萄糖（渗透压对照组）的DMEM培养基中培养。用化学发光法特异性检测细胞内H2O2水平。将细胞用荧光染料CM-H2DCFDA标记后用激光共聚焦显微镜检测活细胞中总ROS水平。用荧光染料Hoechst33258对细胞染色后在荧光显微镜下计数细胞凋亡率。用AnnexinV-FITC/碘化丙锭双染后用流式细胞仪检测各组细胞的凋亡率。在高糖组中同时加入30 mM的抗氧化剂牛磺酸,观察其对细胞凋亡的影响。结果化学发光法检测和共聚焦显微镜检测显示高糖可明显促进肾小管上皮细胞内活性氧的产生,与其他两对照组比较有显著差异（P〈0.01）;Hoechst33258染色法检测和流式细胞仪检测分析显示高糖可明显诱导肾小管上皮细胞的凋亡发生,与其他两对照组比较有显著差异（P〈0.01）;流式细胞仪检测分析显示抗氧化剂牛磺酸可明显抑制高糖诱导的肾小管细胞凋亡,与高糖组比较有显著差异（P〈0.01）。结论高糖可通过促进细胞内生成过量ROS而诱导肾小管细胞凋亡发生,抗氧化剂可阻断高糖诱导的肾小管细胞凋亡。

英文摘要：

Objective To investigate the effect of high glucose on ROS production and apoptosis in the renal tubular epithelial cells and to elucidate the molecular mechanism of renal tubule apoptosis induced by high glucose.Methods Human proximal renal tubular cell line HKC was cultured in DMEM medium containing 5.5 mM D（Dextrorotation）-glucose（normal glucose control）,25 mM D-glucose（high glucose） and 5.5 mM D-glucose ＋ 20 mM L（Laevorotation）-glucose（osmotic pressure control） for different time.Chemoluminescence assay was used to detect H2O2 level intracellularly.ROS（reactive oxygen species） content in HKC was determined with laser confocal microscope after the cells being labeled with fluorescent dye CM-H2DCFDA.After Hoechst 33258 staining,apoptosis rate of the HKC cells was counted under fluorescent microscope.Flow cytometry was used to determine apoptosis rate of HKC cells after dual staining with AnnexinV-FITC/propidium iodide.Meanwhile,30 mM antioxidant taurine was added into medium containing 25 mM D-glucose and its effect on apoptosis of renal tubular cells was observed.Results Chemoluminescence and confocal microscope analyses indicated that high glucose in the medium can promote generation of an excess of ROS in the renal tubular epithelial cells,compared with two other control groups（P〈0.01）.Hoechst 33258 staining revealed that high glucose can obviously increase apoptosis rate in renal tubular cells,compared with two other control groups（P〈0.01）.Flow cytometry analysis showed that taurine can markedly inhibit apoptosis of renal tubular cells induced by high glucose.Conclusion High glucose can induce apoptosis of renal tubular cells by promoting generation of an excess of ROS in human renal tubular cells and taurine can block apoptosis of renal tubular cells induced by high glucose.