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目的观察盐酸吡格列酮对高糖培养的肾小球系膜细胞(mesangial cells,MCs)氧化应激的影响,探讨其肾脏保护机制。方法体外培养MCs随机分组为正常糖(normal glucose group,NG)组、高糖(high glucosegroup,HG)组及高糖+不同浓度吡格列酮(P1、P2、P3)组。培养48 h后,收集各组细胞及上清液。以2’,7’-二氯双氢荧光素二乙酸酯(2’,7’-dichlorofluorescin diacetate,DCFH-DA)为荧光探针,采用流式细胞仪法检测MCs内活性氧(reactive oxygen species,ROS)水平。采用比色法检测MCs上清液中的脂质过氧化物丙二醛(malondialdehyde,MDA)含量和超氧化物歧化酶(superoxide dismutase,SOD)活性。结果与NG组比较,高糖刺激后MCs内ROS水平明显增加,上清液中MDA含量显著增高、SOD活性下降,差异有统计学意义(P<0.001);吡格列酮干预后可明显抑制高糖刺激下的MCs上述变化,且呈浓度依赖性。结论盐酸吡格列酮可抑制高糖诱导的大鼠MCs氧化应激,且呈浓度依赖性,该作用可能部分与其肾脏保护有关。
Objective To observe the effect of pioglitazone hydrochloride on oxidative stress in mesangial cells (MCs) cultured in high glucose and to explore its mechanism of renal protection. Methods MCs cultured in vitro were randomly divided into normal glucose group (NG) group, high glucose group (HG) group and high glucose + pioglitazone group (P1, P2, P3). After 48 h of culture, the cells and supernatants of each group were collected. Flow cytometry was used to detect the reactive oxygen species (ROS) in MCs with 2 ’, 7’-dichlorofluorescin diacetate (DCFH-DA) species, ROS) levels. The contents of malondialdehyde (MDA) and superoxide dismutase (SOD) in supernatant of MCs were measured by colorimetric method. Results Compared with NG group, the level of ROS in MCs increased significantly after MCAO and the content of MDA in supernatant was significantly increased while the activity of SOD decreased significantly (P <0.001). Pioglitazone significantly inhibited the high glucose The above changes of MCs were in a concentration-dependent manner. Conclusion Pioglitazone hydrochloride can inhibit the oxidative stress induced by high glucose in MCs in a concentration-dependent manner, which may be partly related to the protection of kidney.