中国血液净化

›› 2011, Vol. 10 ›› Issue (02): 83-86.doi: 10.3969/j.issn.1671-4091.2011..00

• 基础研究 • 上一篇    下一篇

普罗布考对脂多糖刺激下大鼠腹膜间皮细胞白细胞介素18的表达和对氧化应激的影响

李程程 马健飞 彭 霞 樊 怡 兰海华   

  1. 中国医科大学附属第一医院肾内科
  • 收稿日期:2010-08-02 修回日期:1900-01-01 出版日期:2011-02-12 发布日期:2011-02-12
  • 通讯作者: 马健飞

Effects of probucol on the expression of IL-18 and oxidative stress product in rat peritoneal mesothelial cells stimulated with lipopolysaccharide

LI Cheng-cheng, MA Jian-fei, PENG Xia, FAN Yi, LAN Hai-hua   

  1. Department of Nephrology, the First Hospital of China Medical University, Shenyang 110001, China
  • Received:2010-08-02 Revised:1900-01-01 Online:2011-02-12 Published:2011-02-12

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摘要： 目的 研究普罗布考对脂多糖(lipopolysaccharide,LPS)刺激下大鼠腹膜间皮细胞(rat peritoneal mesothelial cells,RPMC)白细胞介素18(interleukin-18,IL-18)和氧化应激产物丙二醛(malondialdehyde,MDA)表达的影响。 方法 原代培养RPMC，第3代腹膜间皮细胞达80%融合时进入试验。用无血清培养液培养24 h使细胞同步化。随机分组：①正常对照组：只加无血清DMEM/F12培养液；②LPS组：不同浓度LPS(1,10,100 mg/L)培养6 h；10 mg/L LPS分别作用于RPMC 0,3,6,12,24 h；③普罗布考组：20,40,80 μmol/L普罗布考预孵育1 h,加入10 mg/L LPS再作用11 h。用实时聚合酶链反应法检测IL-18 mRNA的表达，酶联免疫吸附法检测细胞上清液中IL-18的蛋白水平，硫代巴比妥酸法检测细胞中MDA的含量。 结果 与正常对照组比较，LPS刺激下RPMC IL-18和MDA的表达明显增加(P＜0.05)，且呈浓度依赖性；随着刺激时间的延长，上述指标呈递增趋势，IL-18于12 h达高峰(P＜0.01)。与10 mg/L LPS组比较，普罗布考可抑制IL-18及MDA的表达(P＜0.05)，随普罗布考浓度增加，抑制作用越强。 结论 LPS作用下RPMC促炎症因子IL-18及氧化应激产物MDA表达增加，普罗布考能抑制上述指标的表达，修复间皮细胞的损伤，改善腹膜炎症状态。

关键词: 脂多糖, 普罗布考, 腹膜间皮细胞, 白细胞介素18, 丙二醛

Abstract: Objective To observe the effects of probucol on the expression of interleukin-18 (IL-18) and oxidative stress product malondialdehyde (MDA) in rat peritoneal mesothelial cells treated with lipopolysaccharide (LPS). Methods Primary culture of rat peritoneal mesothelial cells (RPMCs) were randomly divided into 3 groups: cells growing in serum-free DMEM/F12 medium (normal control group), cells stimulated with different concentrations of LPS (1, 10 and 100mg/L) for 6h or stimulated with 10mg/L LPS for 3, 6, 12 and 24h (LPS group), and cells pretreated with probucol (20, 40 and 80μmol/L) for 1h and then stimulated with 10mg/L LPS for 11h (probucol group). IL-18 mRNA was measured by quantitative real time-PCR, IL-18 in medium was assayed by ELISA, and MDA was determined by thiobarbituric acid method. Results The expressions of IL-18 and MDA increased in RPMCs (P<0.05), and the increases were dependent on the LPS concentration used and the period of LPS stimulation lasted. The peak of IL-18 expression arrived after LPS stimulation for 12h (P<0.01). The expressions of IL-18 and MDA were significantly inhibited in probucol group (P<0.05) as compared with those in LPS group stimulated with 10mg/L LPS, and the inhibition was dependent on the probucol concentration used. Conclusions The expressions of proinflammatory cytokine IL-18 and oxidative stress product MDA were increased in RPMCs stimulated with LPS. Probucol inhibited the two changes, facilitating the repair of cell damage and the reduction of peritoneal inflammation.

Key words: Probucol, Peritoneal mesothelial cell, Interleukin-18, Malondialdehyde