细菌毒素和肿瘤坏死因子-α对人腹膜间皮细胞IL-8和FN分泌的影响.doc.doc

细菌毒素和肿瘤坏死因子-α对人腹膜间皮细胞IL-8和FN分泌的影响 中南大学湘雅医学院附二院 段绍斌 刘伏友 彭佑铭 【摘要】目的：研究细菌毒素和肿瘤坏死因子-α对人腹膜间皮细胞分泌白细胞介素(IL)-8和纤维连接蛋白(FN)的影响。方法：分离人腹膜间皮细胞(HPMC)做体外培养，采用酶联免疫双抗夹心法和逆转录聚酶链反应，检测细胞产生IL-8和FN水平的情况，采用Lowry方法检测培养液中细胞内总蛋白。结果：脂多糖(10μg/ml)、金黄色葡萄球菌肠毒素(10μg/ml)和肿瘤坏死因子-α(TNF-α，1000U/ml) 分别刺激HPMC后，培养液内IL-8和FN蛋白质水平显著增高(P0.01)，并上调IL-8和FN的mRNA表达。结论：细菌毒素和TNF-α可引起人腹膜间皮细胞IL-8和FN蛋白水平增加，并上调HPMCIL-8和FN mRNA表达；FN合成增加可能是导致腹膜纤维化原因之一。 【关键词】腹膜间皮细胞； 细菌毒素； 白细胞介素-8； 纤维连接蛋白 【Abstract】Objective To investigate the effects of bacterial toxin and tumor necrosis factor-α (TNFα) on expression of interleukin-8 (IL-8) and fibronectin (FN) in cultured human peritoneal mesothelial cells (HPMC). Methods Mesothelial cells from human omenta were isolated and cultured in vitro. The quantity of IL-8 and FN in culture supernatants were also determined by ELISA. Reverse transcription and polymerase chain reaction amplification was used to detect the expression of IL-8 mRNA and FN mRNA in HPMC cultured media with or with out bacterial toxin or TNFα. Total cellular protein was estimated in these supernatants using lowry methods. Result The levels of IL-8 and FN in the supernatants were markedly higher, and expression of IL-8 mRNA and FN mRNA were up-regulated. When HPMCs were cultured in the presence of LPS, enterotoxin of staphylococcus aureus or TNFα. Conclusion Bacterial toxin or TNFα can induce the levels of IL-8 and FN to increase, at the same time, they can upregulated the expression of IL-8 and FN mRNA. Improved synthesis of FN may be the one of cause of peritoneal fibrosis. 【Key words】 Peritoneal mesothelial cells; Bacterial toxin; Tumor necrosis factor-α; Interleukin-8; Fibronectin 腹膜是一种生理性半透膜，毛细血管壁、腹膜间质、间皮细胞层、透析液与间皮细胞间及血液与内皮细胞间滞留的液体层是腹膜溶质转运的主要屏障。连续不卧床腹膜透析(CAPD)时，腹膜间皮层长期浸泡在非生理的腹膜透析液中导致腹膜间皮层结构和功能的损害。高渗和酸性腹膜透析液，反复发作的腹膜炎是腹膜纤维化和超滤功能丧失的主要原因。尽管近年来透析技术取得了很大的发展，细菌性腹膜炎仍是CAPD患者死亡的主要原因。腹膜炎发作时，透出液中各种炎性因子如白细胞介素(IL-8)，肿瘤坏死因子(TNFα)及单核细胞趋化因子水平增高[2]。体外实验表明：人腹膜间皮细胞能分泌细胞外基质和IL-8。纤维连接蛋白(FN)是细胞外基质的主要成份，在正常组织中表达较低，在组织损伤炎症反应