氨基胍对乳酸盐腹膜透析液致人腹膜间皮细胞损伤及血管内皮生长因子表达的影响

Effect of aminoguanidine on damage in human peritoneal mesothelial HMr SV5 cells caused by lactate peritoneal dialysis solution andexpression of VEGF

目的研究氨基胍(AG)对乳酸盐腹膜透析液(L-PDS)致人腹膜间皮细胞(HMr SV5)损伤及血管内皮生长因子(VEGF)表达的影响。方法采用人腹膜间皮细胞株HMr SV5为体外实验模型,分为无血清DMEM培养液对照组、L-PDS组(2.5%L-PDS)、L-PDS+AG组(2.5%L-PDS+10 mmol/L AG)、单纯AG组(终浓度10 mmol/L)。各组以上不同干预因素与HMr SV5细胞共孵育。MTT法检测各组细胞增殖、评估细胞活力,Western blot和逆转录-聚合酶链反应(RT-PCR)检测VEGF蛋白及m RNA的表达。结果 MTT试验表明,L-PDS组OD值为(0.120±0.019),明显低于对照组的(0.298±0.031),差异有统计学意义(P<0.05);L-PDS+AG组OD值为(0.289±0.022),明显高于LPDS组,差异有统计学意义(P<0.05)。Western blot和RT-PCR结果均表明,与对照组比较,L-PDS组细胞中VEGF蛋白和m RNA表达明显增加,与L-PDS组比较,PDS+AG组VEGF蛋白和m RNA的表达明显减低,差异均有统计学意义(P<0.05)。结论 AG可拮抗乳酸盐腹膜透析液致人腹膜间皮细胞损伤并下调VEGF的表达。

Objective To investigate the effect of aminoguanidine（AG） on damage in human peritoneal mesothelial cells（HMr SV5） caused by lactate peritoneal dialysis solution（L-PDS） and expression of vascular endothelial growth factor（VEGF）. Methods Depending on the culture of the cell, the cultured HMr SV5 cells were divided to control group（cultured in DMEM）, L-PDS group（2.5% L-PDS）, L-PDS＋AG group（2.5% L-PDS＋10 mmol/L AG）, and AG group（at a final concentration of 10 mmol/L）. MTT assay was used to assess the viability and proliferation of the cells.VEGF protein was determined by western blot, and VEGF m RNA was determined by RT-PCR. Results The optical density（OD） value in L-PDS group was（0.120±0.019）, significantly lower than（0.298±0.031） in the control group（P〈0.05）. The OD value in L-PDS＋AG group was（0.289±0.022）, which was significantly higher than that in L-PDS group（P〈0.05）. Expression of VEGF protein and m RNA increased significantly in L-PDS group as compared with those in control group（P〈0.05）, while the expression decreased significantly in PDS ＋ AG group as compared with those in LPDS group（P〈0.05）. Conclusion AG can antagonize the injury of HMr SV5 caused by L-PDS and down-regulate the expression of VEGF.