Wang Shan, Ye Shan-Dong, Sun Wen-Jia, Hu Yuan-Yuan
Department of Endocrinology, Anhui Provincial Hospital Affiliated to Anhui Medical University, No. 17 Lujiang Road, Hefei 230001, China.
ISRN Endocrinol. 2014 Feb 3;2014:601352. doi: 10.1155/2014/601352. eCollection 2014.
Aim. The purpose of this study was to investigate the effects of pioglitazone on oxidative stress and the expressions of p22(phox) and p47(phox), subunits of NADPH oxidase, in mesangial cells (MCs). Method. Rat mesangial cells were cultured and randomly divided into normal glucose (NG) group, high glucose (HG) group, and pioglitazone group. After 48 h exposure, the supernatants and cells were collected. The expressions of p22(phox) and p47(phox) in MCs were detected by RT-PCR and western blot. The levels of intracellular ROS were determined by flow cytometry. Coloimetry method was used to detect malondialdehyde (MDA) concentrations and superoxide dismutase (SOD) activities. Results. Compared with the NG group, the expression levels of p22(phox), p47(phox) and ROS significantly increased, the activity of SOD decreased in HG group, while the concentration of MDA greatly increased (P < 0.01). Pioglitazone significantly suppressed HG-induced p22(phox) and p47(phox) expressions and oxidative stress. The protein and gene expressions of p22(phox) and p47(phox) were markedly reduced after pioglitazone treatment, so did the ROS generation. The activities of SOD in MCs increased, while the concentrations of MDA in the supernatant decreased greatly by pioglitazone. Conclusions. Pioglitazone can inhibit HG-induced oxidative stress in MCs through suppressing p22(phox) and p47(phox) expressions.
目的。本研究旨在探讨吡格列酮对系膜细胞（MCs）中氧化应激以及烟酰胺腺嘌呤二核苷酸磷酸（NADPH）氧化酶亚基p22（phox）和p47（phox）表达的影响。方法。培养大鼠系膜细胞并随机分为正常葡萄糖（NG）组、高糖（HG）组和吡格列酮组。暴露48小时后，收集上清液和细胞。通过逆转录聚合酶链反应（RT-PCR）和蛋白质印迹法检测MCs中p22（phox）和p47（phox）的表达。采用流式细胞术测定细胞内活性氧（ROS）水平。采用比色法检测丙二醛（MDA）浓度和超氧化物歧化酶（SOD）活性。结果。与NG组相比，HG组中p22（phox）、p47（phox）和ROS的表达水平显著升高，SOD活性降低，而MDA浓度大幅增加（P<0.01）。吡格列酮显著抑制HG诱导的p22（phox）和p47（phox）表达以及氧化应激。吡格列酮处理后，p22（phox）和p47（phox）的蛋白质和基因表达明显降低，ROS生成也降低。吡格列酮使MCs中的SOD活性增加，而上清液中的MDA浓度大幅降低。结论。吡格列酮可通过抑制p22（phox）和p47（phox）表达来抑制HG诱导的MCs氧化应激。
