渗透压制剂对人腹膜间皮细胞透明质酸合成酶的调节作用

Effect of osmotic agents on hyaluronan synthesis in human peritoneal mesothelial cells

目的 :了解不同渗透压制剂对人腹膜间皮细胞 (humanperitonealmesothelialcells,HPMCs)透明质酸合成酶 (hyaluronansynthase ,HAS)的调节作用及对透明质酸 (hyaluronan ,HA)合成的影响。方法 :分离培养的HPMCs随机分为 5组 :正常对照组 (对照组 ) ;90mmol/L葡萄糖组 (高糖组 ) ;30mmol/L葡萄糖组 (低糖组 ) ;5 %缩合葡萄糖 (缩合葡萄糖组 )及 90mmol/L甘露醇组 (甘露醇组 )。给予上述刺激后继续培养 2 4h ,以RT -PCR法检测HAS - 2mRNA和HAS - 3mRNA的表达 ;收集细胞培养上清液 ,以放射免疫法检测HA浓度 ;以微粒排除法观察细胞胞衣样结构的面积。结果 :高糖组、低糖组、缩合葡萄糖组、甘露醇组HAS- 2mRNA的表达均显著高于对照组 (P均 <0 0 5) ;高糖组HAS - 3mRNA的表达亦明显增加 (P <0 0 5) ,低糖组、缩合葡萄糖组、甘露醇组HAS - 3mRNA表达与对照组相比差异无显著 (P >0 0 5)。各实验组细胞胞衣样结构面积与细胞体面积的比值无显著差异 (P均 >0 0 5)。对照组、高糖组、低糖组、缩合葡萄糖组、甘露醇组细胞培养上清液HA的浓度均显著高于对照组 (P均 <0 0 5) ;高糖组HA浓度显著高于低糖组、缩合葡萄糖组和甘露醇组 (P <0 0 5)。结论 :与高浓度葡萄糖相比 ,缩合葡萄糖虽增强HPMCsHAS - 2mRNA表达 ,?

AIM: To understand the effect of osmotic agents on hyaluronan synthesis in human peritoneal mesothelial cells(HPMCs). METHODS: Cultured HPMCs were stimulated with 90 mmol/L glucose(HG), 30 mmol/L glucose(LG), 5% polyglucose(PG) and 90 mmol/L mannitol. Extracellular cell coats were examined using particle exclusion assay, the expressions of hyaluronan synthase 2 and 3(HAS-2 and HAS-3) mRNA in HPMCs were measured by reverse transcription-polymerase chain reaction(RT-PCR). The concentration of hyaluronan in cultured medium was measured by hyaluronan radio-immunoassay kits. RESULTS: HAS-2 mRNA expressions in the four experimental experimental groups were significantly higher than that of the control group. However, only HG enhanced HAS-3 mRNA expression. The sizes of the extracelluar coat in the four experimental groups were not significantly different from that of the control. In addition, hyaluronan concentrations were significantly higher in the four experimental groups as compared with the control group. The hyaluronan concentration in HG group was much higher than that of the LG, PG and mannitol groups. CONCLUSIONS: Although polyglucose increased HAS-2 mRNA expression, it tended to have no effect on HAS-3 mRNA in HPMCs. This result implies that polyglucose might be more biocompatible to peritoneum in peritoneal dialysis as compared with hypertonic glucose.