【摘要】 目的探讨依贝沙坦在造影剂诱导肾小管上皮细胞凋亡中的作用及机制。方法培养的大鼠肾小管细胞(NRK-52E)分别与不同碘浓度(25、50、100、150 mgI／ml)的安射力(非离子型造影剂)共孵育1 h;安射力(100 mgI／ml)先后刺激NRK-52E细胞0．5、1、2、4 h。与安射力(100 mgI／ml)相同渗透浓度的甘露醇(420 mmol／L)与NRK-52E细胞共孵育1 h作为阳性对照。不同浓度依贝沙坦(0．01、0．1、1 mmol／L)先与NRK-52E细胞共孵育1 h后,安射力(100 mgI／ml)刺激1 h。Hoechst染色和流式AnnexinV-FITC／PI双染法检测细胞凋亡。激光共聚焦显微镜检测细胞内活性氧(ROS)水平。RT-PCR检测Bax、Bcl-2 mRNA的表达。结果安射力呈浓度和时间依赖性诱导NRK-52E细胞凋亡。与安射力相同渗透浓度的甘露醇不能明显诱导细胞凋亡。安射力与NRK-52E细胞共孵育后,细胞内ROS产生增多,Bcl-2 mRNA的表达下调。Bax mRNA的表达上调。依贝沙坦呈浓度依赖性抑制安射力诱导的NRK-52E细胞凋亡、细胞内ROS的产生、Bcl-2 mRNA表达下调及Bax mRNA表达上调。细胞内ROS水平与细胞凋亡呈正相关。结论安射力呈浓度和时间依赖性诱导NRK-52E细胞凋亡,此作用与细胞内氧化应激及bcl-2 mRNA表达下调、Bax mRNA表达上调相关。依贝沙坦能抑制上述安射力的作用,呈浓度依赖性抑制NRK-52E细胞凋亡。更多还原

【Abstract】 Objective To investigate the effect and mechanism of irbesartan on the apoptosis of renal tubular cells induced by contrast media. Methods NRK-52E cells were exposed to different concentrations (25, 50, 100, 150 mgI/ml) of ioversol (a non-ionic contrast media)for 1 h. Cells of other groups were incubated with ioversol (100 mgI/ml) for 0.5 h, 1 h, 2 h, 4 h, respectively. Mannitol with the same osmolality as ioversol (420 mmol/L) was used to treat NRK-52E cells as control. In separate experiments, irbesartan (0.01,0.1,1 mmol/L) was added 1 h before incubation with ioversol (100 mgI/ml). Apoptosis was determined by Hoechst stains and flow cytometry AnnexinV-FITC/PI double stains. The intracellular ROS was detected by confocal microscopy with fluorescent probe CM-H2DCFDA. Bax and Bcl-2 mRNA expression was examined by reverse transcription-polymerase chain reaction (RT-PCR). Results Ioversol induced NRK-52E cells apoptosis in a concentration- and time-dependant manner. Mannitol could not induce cell apoptosis. The intracellular ROS generation was markedly increased following ioversol treatment. Furthermore, ioversol induced a decrease in the expression of Bcl-2 mRNA and an increase in the expression of Bax mRNA. Irbesartan attenuated the ioversol-induced apoptosis of NRK-52E cells in a dose-dependent manner, in which the protective effect of irbesartan was dependent on decreasing intracellular ROS formation. In addition, irbesartan reversed the ioversol-induced increase in Bax mRNA and decrease in Bcl-2 mRNA. There was a positive correlation between ROS level and apoptosis rate. Conclusions loversol induces NRK-52E cells apoptosis in a concentration- and time-dependant manner via trigerring oxidative stress and up-regulating the expression of Bax mRNA and down-regulating the expression of Bcl-2 mRNA. Irbesartan can attenuate the above effects of ioversol.更多还原