脂联素对高糖环境下大鼠肾小管上皮细胞血管内皮生长因子表达的影响

Effect of adiponectin on the expression of VEGF in NRK-52E cells induced by high glucose

目的探讨脂联素（adiponectin，ADPN）对高糖环境下大鼠肾小管上皮细胞（NRK-52E）血管内皮生长因子（vascular endothelialgrowthfactor，VEGF）表达的影响。方法用含不同浓度葡萄糖的DMEM培养基体外培养NRK-52E细胞，随机分6组（每组5个样本，重复5次）：A组，含5．5mmol／L葡萄糖培养基对照组；B组，含5．5mmol／L葡萄糖＋甘露醇19．5mmol／L培养基组；C组，含25mmol／L葡萄糖培养基组；D组，含25mmol／L葡萄糖培养基＋脂联素1mg／L组：E组，含25mmol／L葡萄糖培养基＋脂联素2．5mg／L组；F组，含25mmol／L葡萄糖培养基＋脂联素5mc／L组，培养24h。以逆转录一聚合酶链反应（RT-PCR）检测VEGF的表达，免疫荧光检测不同浓度干预组VEGF蛋白的表达影响。结果A组细胞VEGFmRNA表达量为（0．367±0．028）；B组为（0．346±0．045），与A组比较无统计学差异（P〉0．05）；C组为（0．692±0．053），显著高于A组（P〈0．01）；D组为（0．562±0．049），低于C组（P〈0．05）。E组为（0．381±0．035），与A组相近，但无统计学差异（P〉0．05）；F组为（0．295±0．031），低于A组（P〈0．01）。不同浓度脂联素各组间比较，有统计学差异（P〈0．05）。免疫荧光检测VEGF表达情况；B组与A组无差异；C组显著高于A组（P〈0．01）；D组低于C组（P〈0．05）。E组与A组相近，但无统计学差异（P〉0．05）；F组低于A组（P〈0．05）。不同浓度脂联素各组间比较有统计学差异（P〈0．05）。结论ADPN能通过减少肾小管上皮细胞VEGF的表达对肾小管间质起保护作用。

Objective To investigate the effect of adiponeetin （ADPN） on the expression of vascular endothelial growth factor （VEGF） induced by high glucose in kidney tubular epithelial cells （NRK-52E） of rats. Methods The NRK-52E cells were cultured with DMEM medium with different concentrations of glucose for 24 h in vitro in 6 groups （five samples in each group, and the experiment was repeated for 5 times） .. group A （DMEM medium with 5.5 mmol/L of glucose as control group） group B （DMEM medium with 5.5 mmol/L of glucose and 19. 5 mmol/L mannitol） group C （DMEM medium with 25 mmol/L of glucose）, group D （DMEM medium with 25 mmol/L of glucose medium and 1 mg/L of recombinant adiponeetin） ; group E （DMEM medium with 25 mmol/L of glucose medi- um and 2. 5 mg/L of recombinant adiponectin） ; group F （DMEM medium with 25 mmol/L of glucose medium and 5 mg/L of recombinant adiponectin）. The VEGF mRNA was evaluated by RT-PCR and the cellular VEGF was detected by immunofluorescence after 24 h. Results The expression of VEGF mRNA in groups A and B was （0. 367 ± 0. 028） and （0. 346 ±0. 045） respectively （P〈0. 05） ; （0. 692 ± 0. 053） in group C, significantly higher than that in group A （P〈0. 01） ; （0. 562 ± 0. 049） in group D, significantly lower than that in group C （P〈0. 05） ; no significant difference between group E and group A （P〈0. 05）; （0. 295 ±0. 031） in group F, significantly lower than that in group A （P〈0. 05）. There was statistically significant difference bettween different groups of different concentra tions of recombinant adiponectin （P〈0. 05）. The results of immunofluorescence revealed that there was no significant difference in the expression of VEGF between group B and group A. It was significantly higher in group C than in group A （P〈0. 01）, and that in group D was significantly higher than in group C （P〈0. 05）, and there was no significant difference between group E and group A （P〉 0. 05）. The expression of VEGF in group F was significantly lower than in group A （P〈0. 05）. There was statistically significant difference bettween different groups of different concentrations of recombi nant adiponectin （P〈0. 05）. Conclusions ADPN plays a protective role through the alleviation of VEGF expression in renal tubules.