乳酸盐腹膜透析液对人腹膜间皮细胞功能的影响

The effect of lactate-buffered peritoneal dialysis fluid in human peritoneal mesothelial cells function

目的 研究乳酸盐腹膜透析液 (L PDF)对人腹膜间皮细胞 (HPMC)功能的影响。方法 分离HPMC作体外培养 ,以MTT试验测定HPMC增殖程度 ;采用ELISA法检测细胞培养液中白细胞介素 8(IL 8)和纤维连接蛋白 (FN)的蛋白质水平 ;逆转录多聚酶链反应检测IL 8mRNA的表达 ;用Lowry方法检测细胞培养液内总蛋白。结果 L PDF抑制HPMC的增殖 ,且呈时间依赖关系。L PDF刺激HPMC后 ,培养液中IL 8和FN蛋白质水平明显增高 ,并上调IL 8mRNA的表达。在恢复培养期间 ,若加用脂多糖 (10mg/L)、金黄色葡萄球菌肠毒素 (10mg/L)和肿瘤坏死因子 α(TNF α ,10× 10 6U/L)刺激后发现 ,培养液中FN蛋白质水平明显增高 (P <0 .0 5 )。在TNF α刺激组 ,培养液中IL 8水平亦有显著增高 (P <0 .0 1)。结论 乳酸盐腹膜透析液可抑制HPMC增殖 ,并使培养HPMC增高分泌FN和IL 8水平。

Objective To investigate the effect of lactate buffered peritoneal dialysis fluid (L PDF) on human peritoneal mesothelial cells (HPMC) function. Methods Mesothelial cells from human omentum were isolated and cultured in vitro. Proliferation of HPMC was detected by MTT assay. The quantity of interleukin 8 (IL 8) and fibronectin(FN) in culture supernatants was determined by ELISA. Reverse transcription and polymerase chain reaction amplification were used to detect the expression of IL 8 mRNA in HPMC . Total cellular protein in these supernatants was estimated by using Lowry method. Results L PDF inhibited proliferation of HPMC in a time dependentmanner. The levels of IL 8 and FN in the supernatants were remarkably high, and expression of IL 8 mRNA was up regulated in HPMC exposured to L PDF. During a subsequent recovery period of 48 hours in culture medium containing lipopolysaccharide (LPS,10 mg/L), enterotoxin of staphylococcus aureus (10 mg/L) or TNFα(10×10 6 U/L),the levels of FN in the supernatants increased ( P <0.05). The levels of IL 8 also increased in HPMC stimulated with TNFα( P <0.01). Conclusion L PDF inhibited proliferation of HPMC and increased levels of IL 8 and FN in the HPMC.