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目的研究血管内皮细胞生长因子(VEGF)作为促血管生长因子和内抑素(ENS)作为血管生成抑制因子,在糖尿病肾病(DN)血管生成中的作用。方法以正常大鼠为对照,30只雄性Wistar大鼠建成糖尿病模型,分别于糖尿病模型建立2、4、8周观察24小时尿白蛋白排泄率(UAER),用免疫组织化学染色显示大鼠肾组织VEGF和ENS的表达及半定量分析;用酶联免疫法测定血中VEGF和ENS的浓度,根据DN病理学改变确定DN的血管增生严重性,分析内皮细胞增生、肾小球中弥漫或结节病变及肾小球硬化与VEGF和ENS的关系;采用RT-PCR的方法观察大鼠肾脏VEGF和ENS mRNA的表达。结果 DN大鼠肾组织VEGF和ENS的mRNA和蛋白表达水平明显高于对照组(P<0.05);DN大鼠血清VEGF和ENS水平也明显高于对照组(P<0.05)。DN大鼠UAER与血浆VEGF和ENS相关(r=0.468,P<0.01;r=0.395,P<0.05),DN大鼠无论在血中还是肾组织免疫组化和RT-PCR都有VEGF和ENS的表达,VEGF的表达与DN血管增生程度密切相关(r=0.404～0.476,P<0.01～0.05),ENS的表达也与DN严重程度密切相关(r=0.409～0.61 7,P<0.05～0.01),血浆VEGF和ENS二者具有正相关性(r=0.484,P<0.01)。结论 VEGF和ENS与DN血管增生密切相关。
Objective To investigate the role of vascular endothelial growth factor (VEGF) as an angiogenic inhibitor of angiogenic growth factor and endostatin (ENS) in angiogenesis in diabetic nephropathy (DN). Methods Normal rats were used as control. Thirty male Wistar rats were used to establish a diabetic model. Urinary albumin excretion rate (UAER) was measured at 2,4 and 8 weeks after establishment of diabetic model. Immunohistochemical staining showed that rat kidney The expression of VEGF and ENS were detected by semi-quantitative analysis. The concentrations of VEGF and ENS in blood were determined by enzyme-linked immunosorbent assay (ELISA) and the severity of angiogenesis was determined according to the pathological changes of DN. Endothelial cell proliferation, diffuse or knot of glomerulus Pathological changes and glomerular sclerosis and VEGF and ENS; using RT-PCR method to observe the expression of VEGF and ENS mRNA in rat kidney. Results The mRNA and protein expressions of VEGF and ENS in DN rats were significantly higher than those in control rats (P <0.05). The levels of VEGF and ENS in DN rats were also significantly higher than those in control rats (P <0.05). DN rats were associated with plasma VEGF and ENS (r = 0.468, P <0.01; r = 0.395, P <0.05). There was VEGF and ENS in immunohistochemistry and RT- (R = 0.404-0.476, P <0.01-0.05). The expression of ENS was also closely related to the severity of DN (r = 0.409-0.661, P <0.05-0.01) ), There was a positive correlation between plasma VEGF and ENS (r = 0.484, P <0.01). Conclusion VEGF and ENS are closely related to DN hyperplasia.