【摘要】目的 探讨在体外脂多糖(LPS)对大鼠腹膜间皮细胞肿瘤坏死因子- (TNF-)、高迁移率族蛋白B-1(HMGB-1)和转化生长因子1(TGF-1)表达的影响。方法 胰蛋白酶消化法用于PMC的原代培养和传代,经鉴定分组:①正常对照组;②不同浓度LPS组(1、10、100mg/L);③不同时间组:10mg/L LPS 作用于PMC 2、6、12、18、21、24、36h。用RT-PCR法检测HMGB-1mRNA的表达,ELISA法检测细胞培养上清液中TNF-帷MGB-1和TGF-1的表达。 结果 ①LPS诱导PMC HMGB-1 mRNA和蛋白表达,且呈时间浓度依赖;②LPS诱导PMC TNF-岬鞍妆泶锍仕澹虎跿GF-1于2小时表达至高峰随后下降,于12小时表达升高持续至36小时。结论 HMGB-1作为晚期炎症因子参与了腹膜透析相关性腹膜炎的病理过程,LPS通过上调TNF-帷MGB-1和TGF-1的表达,引起持续放大的炎症反应损伤腹膜并导致腹膜纤维化。
【Abstract】 Objective To observe the effect of lipopolysaccharide (LPS) on the expression of tumor necrosis factor alpha (TNF-α), high mobility group box-1 protein (HMGB-1) and transforming growth factor beta-1(TGF-β1) in rat peritoneal mesothelial cell (RPMC). Methods RPMC were isolated from rat colic omentum. The cells were incubated with LPS (1, 10 and 100 mg/L), or were under the stimulation of 10 mg/L LPS for 2, 6, 12, 18, 21, 24 and 36h. RPMC in the control group were incubated with the medium. HMGB-1 mRNA was detected by RT-PCR. TNF-α, HMGB-1 and TGF-β1 were measured by ELISA. Results Compared with the control group, the expression of HMGB-1 mRNA and HMGB-1 increased significantly in the groups stimulated by LPS in time-dependent and dose-dependent manner. TNF-α expression was found to have 2 peaks in the 36h period of LPS stimulation. TGF-β1 increased to the peak at 2h, decreased at 6h, and then increased at 12h to 36h. Conclusion HMGB-1 as a late inflammatory factor may play a role in the pathogenesis of peritonitis relating to CAPD. LPS up-regulates the expression of TNF-α, HMGB-1 and TGF-β1, leading to the continuous amplification of inflammatory processes and the aggravation of peritoneal impairment and fibrosis.