肾缺血再灌注损伤后缺氧诱导因子-1aL及其靶基因miR-210、血管内皮生长因子的动态表达

Dynamic changes of the expression of hypoxia inducible factor-1a and the target genes miR-210, vascular endothelial growth factor in the kidney after ischemic-reperfusion injury

目的观察缺血再灌注损伤后小鼠肾脏缺氧诱导因子（HIF）-1a及其靶基因miR-210、血管内皮生长因子的表达变化，探讨。肾缺血再灌注损伤后血管新生的调控机制。方法将20只成年BALB／C系雄性小鼠随机分为假手术（Sham）组、肾缺血再灌注（IR）4h、1d、3d组，每组5只。各组分别在IR4h、1d、3d后收集血清检测肌酐（Cr）和尿素氮（BUN）的水平，取肾组织标本采用实时定量逆转录-聚合酶链反应（RT—PCR）检测HIF-1amRNA、miR-210、血管内皮生长因子（VEGF）mRNA的表达变化，采用Westernblot检测HIF-1a蛋白的表达变化。结果Cr、BUN和HIF-1amRNA在IR4h、1d组表达水平均明显高于Sham组，分别为（40．20±3．50）、（90．00±3．81）umol／L、（16．88±1．31）、（38．86±5．11）mmol／L，（4．57±0．52）、（3．15±0．46）倍（P〈0．05）；VEGFmRNA、miR-210和HIF—1a蛋白在IR4h、1d、3d组表达水平均明显高于Sham组，分别为[（4．33±0．22）、（3．35±0．22）、（2．51±0．20）倍，（1．29±0．06）、（2．55±0．15）、（1．80±0．13）倍，（1．54±0．08）、（3．08±0．23）、（1．69±0．08）倍，P〈0．05]。结论肾缺血再灌注损伤明显影响HIF—1a及其靶基因miR-210、VEGF的表达，该表达变化可能与肾缺血再灌注损伤血管新生的调控有关。

Objective To Observe the expression of hypoxia inducible factor （HIF）-1a and target genes miR-210, vascular endothelial growth factor （VEGF） and explore the regulatory mechanism of the angiogenesis after ischemia-reperfusion （IR） injury. Methods Twenty BALB/C mice were randomly divided into 4 groups： IR 4 h, 1 day, 3 days groups （IR groups） and sham operation group （sham group）. Four h, 1 day and 3 days after the operation, mice were sacrificed and blood samples were obtained to examine the levels of creatinine （Cr） and urea nitrogen （BUN）. Kidney samples were examined for HIF-1a mRNA, miR-210 and VEGF mRNA by quantitative Real-time reverse transcription-polymerase chain reaction （RT-PCR） , HIF-1 a protein by Western blotting. Results The levels of Cr, BUN and HIF-1 a mRNA in IR 4 h and 1 day groups were significantly increased compared to sham group [ （40. 20± 3.50） and （90.00±3.81） umol/L, （16.88±1.31） and （38.86±5.11） mmol/L, （4.57±0.52） and （3. 15± 0. 46） fold,P 〈 0.05 ]. The levels of miR-210, VEGF mRNA and HIF-1 a protein in IR 4 h, 1 day and 3 days groups were increased apparently compared to sham group [ （4. 33 ± 0. 22 ）, （3.35 ± 0. 22 ）, （2.51 ±0.20） fold, （1.29 ±0.06）, （2.55±0.15）, （1.80±0.13） fold, （1.54 ±0.08）, （3.08 ± 0. 23 ） , （ 1.69 ± 0. 08 ） fold, P 〈 0. 05 ]. Conclusion The renal IR injury can obviously influence the expression of HIF-1 a and target genes miR-210, VEGF, which may be associated with the regulation of the angiogenesis after renal IR injury.