重组人内皮抑素对尿毒症腹膜透析大鼠腹膜新生血管形成的影响

Effect of recombinant human edostatin on peritoneal angiogenesis in uremic peritoneal dialysis rats

目的 研究重组人内皮抑素对尿毒症腹膜透析（PD）大鼠腹膜新生血管形成的影响.方法 40只雄性SD大鼠,按随机数字表法分为正常对照组、肾衰竭非透析组、4.25%PD组、重组人内皮抑素10 mg/kg PD组、重组人内皮抑素40 mg/kg PD组,每组8只.对PD组规律PD 28 d.重组人内皮抑素干预组在行规律PD期间,隔天1次皮下注射重组人内皮抑素,至透析第28天结束.28 d后取各组大鼠新鲜腹膜组织,RT-PCR法检测腹膜组织血管内皮生长因子（VEGF）、碱性成纤维细胞生长因子（bFGF）mRNA表达;免疫组化染色检测VEGF、bFGF蛋白表达.CD34染色观察腹膜组织毛细血管密度（MVD）.结果 各组大鼠腹膜组织均表达VEGF和bFGF,肾衰竭非透析组、4.25%PD组VEGF及bFGF mRNA、蛋白表达均显著高于正常对照组（均P＜0.05）;重组人内皮抑素10 mg/kg PD组、40 mg/kg PD组VEGF及bFGF mRNA、蛋白表达均显著低于4.25%PD组（均P＜0.05）.肾衰竭非透析组、4.25%PD组腹膜组织MVD均显著高于正常对照组（均P＜0.05）;重组人内皮抑素10 mg/kg、40 mg/kg PD组腹膜组织MVD均显著低于正常对照组（均P＜0.05）.结论 重组人内皮抑素可以有效抑制PD大鼠腹膜新生血管的形成,下调VEGF、bFGF mRNA及蛋白表达可能是其抑制腹膜新生血管形成的机制之一.

Objective To study the effect of recombinant human edostatin on peritoneal angiogenesis in uremic peritoneal dialysis（PD） rats. Methods Forty male SD rats were randomly divided into 5 groups： normal control rats （group 1）, renal failure without PD rats （group 2）, rats dialyzed with 4.25% PD solution （group 3）, rats dialyzed with 4.25% PD solution and received subcutaneous injection of recombinant human endostatin 10 mg/kg （group 4）, rats dialyzed with 4.25% PD solution and received subcutaneous injection of recombinant human endostatin 40 mg/kg （group 5）. Recombinant human endostatin was given every other day during peritoneal dialysis period, total 14 times. After regular PD for 28 days, tissue immunohistochemical staining and RT-PCR were used to detect the mRNA and protein expressions of VEGF and bFGF in peritoneal tissues of each group rats. Microvessel density （MVD） of peritoneum was detected and quantified with anti-CD34 immunohistochemical staining. Results The mRNA and protein of VEGF and bFGF were expressed in each group. Compared to group 1, the mRNA and protein expression of VEGF and bFGF were significantly up-regulated in group 2 and group 3 （all P〈0.05）. Compared with group 3, the mRNA and protein expression of VEGF and bFGF were significantly downregulated in group 4 and group 5 （all P〈0.05）. Compared with group 4, the mRNA and protein expression of VEGF and bFGF were significantly down-regulated in group 5 （all P〈0.05）. The new microvascular vessels in group 1 showed little or none. Compared with group 1, MVD was significantly increased in group 2 and group 3 （P〈0.05）. Compared with group 3, MVD was significantly decreased in group 4 and group 5 （all P〈0.05）. Conclusions Recombinant human endostatin can effectively inhibit rat peritoneal neoangiogensis. Down-regulated expression of VEGF and bFGF in peritoneum may be one of the mechanisms of recombinant human endostatin inhibiting peritoneal angiogenesis.