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。
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.
