摘要
Objective To observe the influence of glucose on the expression of glucose transporters (GluTs) in peritoneal tissues.Methods Mesothelial cells (MsCs) from Sprague-Dawley (SD) rats were cultured in medium with glucose 214.4 mmol/L or 75.5 mmol/L. The normal medium with glucose 17.5 mmol/L was used as control. Total RNA was extracted from each sample after 24 hours incubation. Reverse transcript polymerase chain reaction (RT-PCR) was performed with primers corresponding to sodium-glucose transporter (SGIT1) and GluT1 -GluT4. mRNA expression of the above GluTs from each sample was measured with quantitative PCR.Results GluT1 and GluT2 mRNA can be detected in MsCs from SD rats, while no positive bands can be found specificaly for GluT3, GluT4 and SGIT1. Quantitating the amount of PCR products indicated that the levels of GluT1 mRNA in MsCs cultured 24 h in both 214.4 mmol/L glucose and 75.5 mmol/L glucose medium decreased dramatically compared with that in normal medium ( P≤0.01 ). While under the same conditions, the levels of GluT2 mRNA in MsCs cultured 24 h in 214.4 mmol/L and 75.5 mmol/L glucose medium both increased significantly ( P ＜ 0.01 ).Conclusions GluT1 is strongly expressed in MsCs under normal glucose levels and decreased dramatically under high glucose conditions, while GluT2 expressed at a low level in normal medium and increased greatly after incubation in high glucose conditions. This may play a great role in glucose absorportion during peritoneal dialysis and have some connection with ultrafiltration failure due to the alteration of glucose absorption after long-term dialysis.
目的 长期腹膜透析后腹膜对葡萄糖的吸收及腹膜溶质转运特点常会发生改变 ,这可能与腹膜组织葡萄糖载体(GlucoseTransporters,GluTs)表达改变有关。本研究旨在观察腹膜间皮细胞GluTs表达情况及不同浓度葡萄培养对腹膜间皮细胞GluTs表达的影响。方法 分别采用高糖 (含葡萄糖 2 14 4mmol L)及低糖 (含葡萄糖 75 5mmol L)培养基 ,并用正常培养基 (含葡萄糖17 5mmol L)作为对照 ,体外培养SD大鼠腹膜间皮细胞。 2 4小时后抽提细胞总RNA ,逆转录成cDNA后用GluTs各亚型SGlT1及GluT1～GluT4相对应的引物进行PCR扩增。采用以自身cDNA为定量标准的RT PCR方法 ,对各标本内GluTs各亚型mRNA表达量进行半定量分析。结果 体外培养的大鼠腹膜间皮细胞存在GluT1及GluT2mRNA的阳性表达 ,未见到SGlT1、GluT3及GluT4mRNA的阳性表达条带。用含葡萄糖 2 14 4mmol L及 75 5mmol L的培养基较用正常培养基培养 ,腹膜间皮细胞GluT1mRNA的表达显著减少 (P <0 0 1) ;而在相同培养条件下GluT2mRNA的表达量明显增加 (P <0 0 1)。结论 在接近生理性葡萄糖浓度下 ,腹膜间皮细胞高效表达GluT1但低表达GluT2 ,但在高浓度葡萄糖的环境中 ,GluT1表达明显下降而GluT2则出现高效表达。
基金
theKeyProjectFoundationoftheMinistryofPublicHealth (No 970 40 2 2 8)
