炎症介质对人腹膜间皮细胞透明质酸合成酶mRNA表达和透明质酸合成的影响

Effects of inflammation mediators on the mRNA expression of hyaluronan synthases and hyaluronan synthesis in human peritoneal mesothelial cells

目的 了解炎症介质对人腹膜间皮细胞(HPMCs)透明质酸合成酶mRNA的调节作用及对透明质酸合成的影响。方法 分离培养的人腹膜间皮细胞随机分为4组:正常对照组、脂多糖组(LPs,10 ng/ml)、肿瘤坏死因子-α(TNF-α,100 U/ml)组,白介素-1β(IL-1β,100 U/ml)组,给予上述刺激后继续培养24 h。人腹膜间皮细胞HAS-2及HAS-3 mRNA表达以逆转录多聚酶链反应(RT-PCR)法检测;细胞衣样结构合成以微粒排除法检测;细胞培养上清液透明质酸(HA)的浓度以放射免疫分折法检测。结果 LPS组、TNF-α组HAS-2 mRNA表达分别较对照组增强1.2倍和1.3倍(P均<0.05);IL-1β组HAS-2 mRNA表达虽较对照组增强,但差异无显著性意义(P>0.05)。LPS组、1L～1β组、TNF-α组HAS-3 mRNA表达分别较对照组增强1.7倍、19倍、8.5倍(P均<0.05)。对照组、LPS组、IL-1β组、TNF-α组细胞胞衣样结构面积与细胞体面积的比值各组间差异无显著性意义(P>0.05);对照组、LPS组、IL-1β组、TNF-α组细胞培养上清液透明质酸(HA)的浓度均显著高于对照组(P均<0.05)。结论炎症因子可上调HPMCs透明质酸合成酶HAS-2 mRNA、HAS-3 mRNA的表达和促进透明质酸的合成。由于其主要增强HAS-3 mRNA的表达,提示炎症状态时有大量小相对分子质量透明质酸合成。

Objective To elucidate the effect of inflammation mediators on the mRNA expression of hyaluronan synthases and hyalaronan synthesis in human peritoneal mesothelial cells(HPMCs) . Methods Cultured HPMCs were stimulated with LPS(10 ng/ml), TNF-α(100 U/ml) and IL-1β(100 U/ml). The expression of hyaluronan synthase 2 and 3(HAS2 and HAS3) mRNA in HPMCs was measured by reverse transcription-polymerase chain reaction(RT-PCR) . The extracellular cell coats were observed using particle exclusion assay, and the concentration of hyaluronan in cultured medium was measured by hyaluronan radio-immunoassay kits. Results The expression of HAS-2 mRNA in LPS and TNF-α group was respectively 1. 2 and 1. 3 fold higher as compared with the control group. However, the HAS-2 mRNA expression in IL-1β group did not increase significantly. The levels of HAS-3 mRNA expression in these inflammatory mediator groups were respectively 1. 7, 8. 5, and 19 fold higher as compared with the control group. However, the sizes of the pericelluar hyaluronan coats in HPMCs did not change significantly after LPS, IL-1β or TNF-α stimulation. The concentrations of hyaluronan in inflammation mediator groups were significantly higher than that of control group. Conclusions Inflammatory mediators increase both hyaluronan sythases mRNA expression and the hyaluronan synthesis. Their effects on HAS-3 expression were much stronger than those on HAS-2 expression. These data indicate that during inflammation HPMCs may synthesized lots of low molecular weight hyaluronan.