论文部分内容阅读
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,设立正常对照(NC)组、高糖(HG)组、高糖+不同浓度吡格列酮(PIO1、PIO2、PIO3)组。以荧光探针2′,7′-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内活性氧簇(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 48h后,细胞内ROS水平升高,MCs PAI-1mRNA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48h后,PIO1、PIO2、PIO3组细胞内ROS水平降低,MCs PAI-1mRNA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-1mRNA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,并呈一定的浓度依赖性,该作用可能部分与其肾脏保护有关。
Objective To observe the effect of pioglitazone on oxidative stress and plasminogen activator inhibitor 1 (PAI-1) expression in rat mesangial cells (MCs) stimulated by high glucose and explore its mechanism of renal protection. Methods Rat glomerular MCs were cultured in vitro. Normal control (NC) group, high glucose (HG) group, high glucose + pioglitazone (PIO1, PIO2, PIO3) groups were established. The cells were labeled with fluorescent probe 2 ’, 7’-dichlorodihydrofluorescein diacetate (DCFH-DA), and the level of reactive oxygen species (ROS) in MCs was detected by flow cytometry; PAI -1 mRNA expression in supernatant of culture supernatant PAI-1 protein by ELISA. Results Compared with NC group, the intracellular ROS level increased and the expression of PAI-1mRNA and PAI-1 protein in MCs increased after 48h stimulation with high glucose. Compared with HG group, after 48h intervention with Pioglitazone, PIO1 PIO2 and PIO3 decreased the level of ROS, the expression of PAI-1 mRNA in MCs and the concentration of PAI-1 in supernatant decreased in a concentration-dependent manner. Correlation analysis showed that ROS levels in MCs were positively correlated with PAI-1mRNA expression and PAI-1 protein concentration in supernatant. Conclusion Pioglitazone can inhibit the high glucose-induced glomerular MCs ROS generation and PAI-1 overexpression, and in a concentration-dependent manner, this effect may be partly related to its renal protection.