·糖尿病基础研究·

　　【摘要】 目的 观察吡格列酮对高糖刺激下大鼠肾小球系膜细胞(MCs)氧化应激和纤溶酶原激活物抑制物1(PAI-1)表达的影响，探讨其肾脏保护机制。 方法 体外培养大鼠肾小球MCs，设正常对照(NC)组、高糖(HG)组、高糖+不同浓度吡格列酮组(PIO1、PIO2、PIO3组)。以荧光探针2ˊ，7ˊ-二氯双氢荧光素二乙酸酯(DCFH-DA)标记细胞，采用流式细胞仪检测MCs内活性氧(ROS)水平;以RT-PCR检测PAI-1的mRNA表达;采用ELISA检测培养细胞上清液PAI-1蛋白浓度。结果 与NC组相比，高糖刺激MCs 48 h后，细胞内ROS水平升高，MCs PAI-1 mRNA表达和上清液PAI-1蛋白浓度增高;与HG组相比，经吡格列酮干预48 h后，PIO1、PIO2、PIO3组细胞内ROS水平降低，MCs PAI-1 mRNA表达和上清液PAI-1蛋白浓度降低，并呈一定的浓度依赖性。相关性分析显示，MCs内ROS水平与PAI-1 mRNA表达、上清液PAI-1蛋白浓度均呈正相关。结论 吡格列酮可抑制高糖诱导的大鼠肾小球MCs ROS生成和PAI-1过度表达并呈一定的剂量依赖性，该作用可能部分与其肾脏保护有关。

　　【关键词】 氧化应激;纤溶酶原激活物抑制物1;系膜细胞;吡格列酮;高糖

Effect of pioglitazone on inhibiting high glucose mediated oxidative stress and plasminogen activator inhibitor-1 expression of rat glomerular mesangial cells in vitro DONG Chong-zhou, YE Shan-dong, HUANG Hai-quan, et al. Department of Endocrinology, Tongling People’s Hospital, Tongling 244000, China

Corresponding author: YE Shan-dong, E-mail: ysd6464@yahoo.com.cn

　　【Abstract】Objective To observe the effect of pioglitazone on inhibiting high glucose mediated oxidative stress and plasminogen activator inhibitor-1 (PAI-1) expression of rat glomerular mesangial cells in vitro and to explore the mechanism of its reno-protection. Methods The rat mesangial cells were cultured in the medium with normal glucose (NC group ), high glucose (HG group) and high glucose plus different concentrations of pioglitazone (PIO1,PIO2 and PIO3 group respectively). The intracellular reactive oxygen species (ROS) level was detected by flow cytometry, using 2ˊ, 7ˊ-dichlorofluorescein diacetate (DCFH-DA) as a ROS fluorescent dye. The expression of PAI-1 mRNA was detected by RT-PCR. The level of PAI-1 protein in the supernatants was determined by ELISA. Results Compared with the NC group, the intracellular ROS content, PAI-1 mRNA expression of mesangial cells, and PAI-1 protein level in the supernatants increased significantly in the HG group. Compared with the HG group, after the intervention with pioglitazone for 48h, there was a significant reduction in the intracellular ROS content, PAI-1 mRNA expression and PAI-1 protein level in PIO1, PIO2 and PIO3 groups in a concentration-dependent manner. The correlation analysis showed that the intracellular ROS content in mesangial cells was positively correlated with both PAI-1 mRNA expression and PAI-1 protein level. Conclusion Pioglitazone can inhibit high glucose-induced ROS and PAI-1 expression of mesangial cells in a dose-dependent manner, which may contribute to its reno-protection partly.

　　【Key words】 Oxidative stress; Plasminogen activator inhibitor-1 (PAI-1); Mesangial cells (MCs); Pioglitazone; High glucose