Chen J Y, Chiu J H, Chen H L, Chen T W, Yang W C, Yang A H
Department of Medicine, Taipei Veterans General Hospital, Taiwan.
Perit Dial Int. 2000 Nov-Dec;20(6):772-7.
To investigate the induction of nitric oxide synthase type II (iNOS) in human peritoneal mesothelial cells (HPMC) using cytokines and bacterial lipopolysaccharide (LPS).
Confluent monolayers of HPMC were exposed to cytokines [tumor necrosis factor alpha (TNFalpha), interleukin-1 beta (IL-1beta), interferon gamma (IFNgamma)] or LPS, individually or in various double and triple combinations, for 24-72 hours. Concentrations of nitrate and nitrite in the media were quantified using the Griess reaction and used as indirect indices of nitric oxide (NO) production. The expression of iNOS was assessed using reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot.
Neither single cytokines nor LPS was able to induce iNOS mRNA or NO production. Both double combinations of TNFalpha + IFNgamma and IL-1beta + IFNgamma were able to induce iNOS mRNA expression, but only TNFalpha + IFNgamma induced significant NO production. The triple combination of TNFalpha + IFNgamma + IL-1beta induced even more NO production than TNFalpha + IFNgamma. There was no constitutive NO synthase type III (eNOS) expression in HPMC.
Certain combinations of cytokines could stimulate cultured HPMC to produce NO, and HPMC might be a source of intraperitoneal NO production during peritonitis.
利用细胞因子和细菌脂多糖（LPS）研究人腹膜间皮细胞（HPMC）中诱导型一氧化氮合酶（iNOS）的诱导情况。
将融合的HPMC单层细胞分别或多种双重和三重组合形式暴露于细胞因子[肿瘤坏死因子α（TNFα）、白细胞介素-1β（IL-1β）、干扰素γ（IFNγ）]或LPS中24至72小时。使用格里斯反应对培养基中的硝酸盐和亚硝酸盐浓度进行定量，并将其用作一氧化氮（NO）产生的间接指标。使用逆转录聚合酶链反应（RT-PCR）和蛋白质免疫印迹法评估iNOS的表达。
单一细胞因子或LPS均不能诱导iNOS mRNA或NO产生。TNFα + IFNγ和IL-1β + IFNγ的双重组合均能诱导iNOS mRNA表达，但只有TNFα + IFNγ能诱导显著的NO产生。TNFα + IFNγ + IL-1β的三重组合诱导产生的NO甚至比TNFα + IFNγ更多。HPMC中不存在组成型一氧化氮合酶III（eNOS）表达。
某些细胞因子组合可刺激培养的HPMC产生NO，且HPMC可能是腹膜炎期间腹膜内NO产生的来源。
