缺血再灌注损伤对小鼠肾脏血管内皮生长因子受体3表达的影响

Expression of Vascular Endothelial Growth Factor Receptor3 in the Mice Kidneys with Ischemia Reperfusion Injury

研究肾脏缺血再灌注（ischemia．reperfusion，IR）对小鼠。肾脏血管内皮生长因子受体3（vascular endothelial growth factor receptor3，VEGFR3）表达的影响。方法30只雄性C57BL／6小鼠随即分为5组：假手术组（Sham组）、缺血再灌注0、6、12、24h组（IR0h组、IR6h组、IR12h组、IR24h组），每组6只。缺血再灌注组用无创性动脉夹夹闭左侧肾带，置于32℃温箱后1h松开血管夹，随后切除右肾。Sham组操作同上。但不夹闭左侧。肾蒂。再灌注0、6、12、24h后处死小鼠，收集肾脏及小鼠外周血标本。测定血肌酐（cr）和尿素氮（BUN）水平。PAS染色后显微镜下观察肾脏病理学变化，Western印迹和免疫组织化学法检测肾脏组织血管内皮生长因子受体3的表达。结果与Sham组相比较，再灌注0h时小鼠血肌酐和尿素氮无明显升高，但缺血再灌注6、12、24h后血肌酐和尿素氮呈显著性上升。IR各组肾组织病理损伤也随着再灌注时间的延长而逐渐加重，可见肾小管上皮细胞明显肿胀坏死，蛋白管型形成，刷状缘脱落。随着缺血再灌注时间的进展，VEGFR一3蛋白表达量增加，免疫组织化学染色结果显示VEGFR3主要分布在肾脏皮质髓质交界处的肾小管。结论VEGFR3在。肾脏缺血再灌注损伤后表达量上升，且表达部位主要分布于肾脏皮髓质交界处的肾小管，因此VEGFR3可能参与调控肾脏缺血再灌注损伤。

Objective To investigate the effect of renal ischemia-reperfusion （IR） on the ex- pression of vascular endothelial growth factor receptor 3 （ VEGFR3 ） . Methods Thirty male C57/ BI/5 mice were randomly divided into five groups： Sham group and IR groups （IR 0-h group, IR 6-h group, IR12-h group and IR 24-h group） with 6 mice in each group. The left renal artery of mice in IR groups was occluded with the noninvasive arterial clip and the mice were then kept in a 32~C incubator for 1 h, and had their kidneys removed thereafter. Mice in the sham group experienced the same procedure except that the left renal artery was not clamped. Animals were sacrificed after reperfusion for 0, 6, 12, or 24 h. The kidney and blood samples were collected. The levels of serum creatinine （Cr） and urea nitrogen （BUN） were detected. The pathological changes of renal tissues were observed by PAS staining. The expression of VEGFR3 was detected by Western blotting and immunohistochemistry, respectively. Results The levels of Cr and BUN in IR 0-h group were not significantly different from those in the sham group, but they were significantly increased in a time-dependent manner after IR for 6, 12 and 24 h. Renal pathological injury was significantly ag- gravated with prolonged reperfusion duration. There were obvious swelling and necrosis of renal epithelial cells. Proteincasts developed. The renal brush border dropped off. The expression of VEGFR3 was increased as the IR time increased. Immunohistochemistry revealed that VEGFR3 was predomi- nantly expressed in the renal tubule at the junction of the renal cortex and medulla. Conclusion The expression of VEGFR3, mainly distributed in the renal tubule at the junction of the renal cortex and medulla, is increased after renal IR injury. VEGFR3 may participate in the renal IR injury.