摘要
目的研究肾毒血清对骨髓内皮祖细胞(EPCs)增生及血管内皮生长因子表达的影响。方法分离SD大鼠骨髓内皮祖细胞,将其分为对照组和肾毒血清组[后者按所含肾毒血清的不同体积浓度(1.25%、2.5%、5.0%)分3个亚组,肾毒血清取自成模12周后的5/6肾切除模型大鼠];利用MTT检测EPCs的增生率,ELISA检测培养上清VEGF的浓度,RT-PCR检测VEGFmRNA的表达。结果不同浓度的肾毒血清在不同时间点对培养EPCs均有不同程度的增生抑制作用(除1.25%的肾毒血清在12h时外,其他均P<0.01);同一浓度的肾毒血清对EPCs的增生抑制作用随着刺激时间的延长而增加,24h的增生率显著低于12h(均P<0.01),48h的增生率显著低于24h(均P<0.01);12h时,各肾毒血清组EPCs分泌的VEGF均显著高于对照组(均P<0.01),24h时,5.0%肾毒血清组EPCs分泌的VEGF则显著低于对照组(P<0.01);48h时,各肾毒血清组EPCs分泌的VEGF均低于对照组。对照组12h、24h和48h时VEGFmRNA表达分别为(0.55±0.03)、(0.53±0.02)和(0.49±0.04),2.5%肾毒血清组12h、24h和48h时VEGFmRNA表达分别为(0.50±0.02)、(0.34±0.02)和(0.15±0.01),2.5%肾毒血清组各时间点之间的VEGF差异有显著性(均P<0.05)。结论肾毒血清能抑制EPCs的增生可能是通过抑制EPCs的VEGFmRNA表达、进而减少VEGF分泌而实现的。
[Objective] To study the effect of renal toxic serum on bone marrow endothelial progenitor cell proliferation and the expression of vascular endothelial growth factor.[Methods] Bone marrow endothelial progenitor cells（EPCs） were separated from SD rats,which were divided into the control and renal toxic serum group（according to the volume concentration the latter was divided into 1.25%,2.5% and 5.0% three subgroups,and renal toxic serum was obtained from a 12 weeks 5/6 nephrectomy rat model）.The viability of EPCs was assessed by MTT assay.VEGF concentration of supernatant was determined by ELISA.A reverse transcription polymerase chain reaction（RT-PCR） was used to detect the expression of VEGF mRNA.[Results] Different concentrations of renal toxic serum inhibited EPCs proliferation at different time points（P 0.01,except the concentration of 1.25% on the point of 12 h）;and the same concentration of nephrotoxic serum induced a significant time-dependent loss of cell viability.EPCs proliferation rate in different concentrations of renal toxic serum group at 24 h was significantly lower than that at 12 h（all P 0.01）,EPCs proliferation rate at 48 h was significantly lower than that at 24 h and 12 h（all P 0.01）.VEGF concentrations in renal toxic serum group at 12 h were significantly higher than those in control group（all P 0.01）,VEGF concentration in 5% renal toxic serum group at 24 h was significantly lower than that in control group（all P 0.01）,VEGF concentrations in every renal toxic serum group at 48 h were significantly lower than those in control group（all P 0.01）.VEGF mRNA expression in control group at 12 h,24 h,48 h were [（0.55±0.03）,（0.53±0.02）,（0.49±0.04）] respectively,VEGF mRNA expression in 2.5% nephrotoxic serum group at 12 h,24 h,48 h were [（0.50±0.02）,（0.34±0.02）,（0.15±0.01）] respectively,which were significant different at different time point（all P 0.05）.[Conclusion] Nephrotoxic serum can inhibit the proliferation of EPCs by lowering the expression of VEGF mRNA,thereby reducing secretion of VEGF achieved.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2011年第16期1836-1840,共5页
China Journal of Modern Medicine
基金
国家自然科学基金项目(No:30500546)
关键词
内皮祖细胞
肾
血清
血管内皮生长因子
大鼠
endothelial progenitor cells
renal
serum
vascular endothelial growth factor
rats
