Tristetraprolin和血管内皮生长因子家族的表达异常参与腹膜透析大鼠超滤衰竭的发生  

The abnormal expressions of tristetraprolin and vascular endothelial growth factor family which participate in ultrafiltration failure

作　　者：肖静[1] 高欢欢[1] 赵志红[1] 靳云凤[1] 王晓阳[1] 刘栋[1] 赵占正[1] 刘章锁[1] 

机构地区：[1]郑州大学第一附属医院肾脏内科, 450052

出　　处：《中华肾脏病杂志》2014年第7期512-518,共7页Chinese Journal of Nephrology

摘　　要：目的 观察尿毒症大鼠腹膜组织tristetraprolin （TTP）与血管内皮生长因子（VEGF）家族表达及淋巴管新生情况,探讨TTP与淋巴管新生是否参与了超滤衰竭（UFF）的发生.方法 选取清洁级健康SD大鼠40只（180 ～ 200 g）,采用随机区组法分为5组：正常对照组、假手术组、尿毒症组、PD2周组和PD4周组,每组8只.应用5/6大鼠肾脏切除法构建尿毒症大鼠模型,并在此基础上建立腹膜透析大鼠模型.PD组行规律腹膜透析2周或4周.正常对照组、PD2周组、PD4周组SD大鼠于处死前,行腹膜平衡实验（PET）,计算葡萄糖转运量和超滤量.RT-PCR法检测腹膜组织VEGF、VEGF-A、VEGF-B、VEGF-C、VEGF-D、TTP mRNA的表达,免疫组化或Western印迹法检测腹膜中VEGF、VEGF-C、TTP蛋白的表达,CD31、LYVE-1分别标记腹膜组织新生血管密度（MVD）和淋巴管密度（LVD）.结果 （1）与正常对照组相比,PD2周组和PD4周组葡萄糖转运量均增加（P＜0.05）,超滤量均减少（P＜0.05）,且PD4周组较PD2周组更显著（P＜0.05）.（2）与正常对照组相比,尿毒症组和PD4周组大鼠腹膜的CD31、LYVE-1蛋白表达均增加（P＜0.05）,且腹膜的MVD和LVD亦增加（P＜0.05）;与尿毒症组相比,PD4周组更显著（P＜0.05）.（3）与正常对照组相比,尿毒症组大鼠VEGF、VEGF-A、VEGF-B、VEGF-C、VEGF-D mRNA表达均增加（P＜0.05）,且PD4周组的表达量显著高于尿毒症组（P＜0.05）.其中PD4周组的VEGF和VEGF-C mRNA和蛋白表达均高于PD2周组（P＜0.05）.（4）与正常对照组相比,尿毒症组和PD2周组TTP蛋白表达减少（P＜0.05）,而与尿毒症组和PD2周组比较,PD4周组大鼠TTP蛋白表达显著减少（P＜0.05）.结论 在机体内尿毒症和高糖腹透液环境下TTP表达异常,使其对VEGF家族表达失调控,且具有腹透时间依赖性,进而促进超滤衰竭的发生.高糖腹透液导致血管和淋巴管的新生,二者均参与超滤衰竭的发生.Objective To observe the expression of tristetraprolin （TTP） and vascular endothelial growth factor （VEGF） family,to test and verify whether lymphangiogenesis was involved in the occurrence of ultrafiltration failure （UFF） as well as angiogenesis.Methods Forty male SD rats of clean grade were selected （180-200 g）.These rats were divided into five groups randomly：normal group （n=8）,sham operation group （n=8）,uremia group （n=8）,peritoneal dialysis （PD） 2-week group （n=8）,PD 4-week group （n=8）.The uremic rats model was established by 5/6 nephrectomy,and of which the PD rats model was established on the basis.The rats of PD2-week group and PD4-week group were given regular PD with 4.25% peritoneal dialysis fluid in dose of 3 ml/100 g body weight.PD2-week group received peritoneal dialysis for 2 weeks,PD4-week group for 4 weeks.Before the rats were sacrificed,peritoneal equilibration test （PET） was applied to calculate the mass transfer of glucose and peritoneal ultrafiltration volume.The protein expressions of VEGF,VEGF-C in each group of rats＇ parietal peritoneum were detected by immunohistochemical staining.Microvessel density （MVD） and lymphatic vessel density （LVD） of peritoneal tissue were marked and quantified with anti-CD31 antibody,anti-LYVE-1 antibody.RT-PCR was applied to detect the mRNA expressions of VEGF-A,VEGF-B,VEGF-C,VEGF-D,TTP.Western blotting was used to detect the protein expression of TTP.Results （1）PET revealed that,compared with normal group,the mass transport of glucose and the peritoneal ultrafiltration volume of both PD 2-week group and PD 4-week group elevated （P 〈 0.05）; and compared with PD 2-week group,PD 4-week group＇s elevated （P 〈 0.05）.（2） Compared with normal group,the protein expression of CD31,LYVE-1,the count of MVD and LVD were increased in uremia group and PD4-week group （P 〈 0.05）.Those of PD4-week groups likewise were increased compared to uremia group （P 〈 0.05）.（3） Compared with nor

关 键 词：Tristetraprolin蛋白 腹膜透析 超滤 新生血管化 病理性 淋巴管新生 血管内皮生长因子 

分 类 号：R692.5[医药卫生—泌尿科学]