CAJ | 학술논문

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.
목적 관찰뇨독증대서복막조직tristetraprolin (TTP)여혈관내피생장인자(VEGF)가족표체급림파관신생정황,탐토TTP여림파관신생시부삼여료초려쇠갈(UFF)적발생.방법 선취청길급건강SD대서40지(180 ～ 200 g),채용수궤구조법분위5조:정상대조조、가수술조、뇨독증조、PD2주조화PD4주조,매조8지.응용5/6대서신장절제법구건뇨독증대서모형,병재차기출상건립복막투석대서모형.PD조행규률복막투석2주혹4주.정상대조조、PD2주조、PD4주조SD대서우처사전,행복막평형실험(PET),계산포도당전운량화초려량.RT-PCR법검측복막조직VEGF、VEGF-A、VEGF-B、VEGF-C、VEGF-D、TTP mRNA적표체,면역조화혹Western인적법검측복막중VEGF、VEGF-C、TTP단백적표체,CD31、LYVE-1분별표기복막조직신생혈관밀도(MVD)화림파관밀도(LVD).결과 (1)여정상대조조상비,PD2주조화PD4주조포도당전운량균증가(P＜0.05),초려량균감소(P＜0.05),차PD4주조교PD2주조경현저(P＜0.05).(2)여정상대조조상비,뇨독증조화PD4주조대서복막적CD31、LYVE-1단백표체균증가(P＜0.05),차복막적MVD화LVD역증가(P＜0.05);여뇨독증조상비,PD4주조경현저(P＜0.05).(3)여정상대조조상비,뇨독증조대서VEGF、VEGF-A、VEGF-B、VEGF-C、VEGF-D mRNA표체균증가(P＜0.05),차PD4주조적표체량현저고우뇨독증조(P＜0.05).기중PD4주조적VEGF화VEGF-C mRNA화단백표체균고우PD2주조(P＜0.05).(4)여정상대조조상비,뇨독증조화PD2주조TTP단백표체감소(P＜0.05),이여뇨독증조화PD2주조비교,PD4주조대서TTP단백표체현저감소(P＜0.05).결론 재궤체내뇨독증화고당복투액배경하TTP표체이상,사기대VEGF가족표체실조공,차구유복투시간의뢰성,진이촉진초려쇠갈적발생.고당복투액도치혈관화림파관적신생,이자균삼여초려쇠갈적발생.
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 normal group,the mRNA expressions of VEGF,VEGF-A,VEGF-B,VEGF-C,VEGF-D were significantly increased in uremia group (P ＜ 0.05); Compared with uremia group,the expressions in PD4-week group were significantly increased (P ＜ 0.05).Compared with normal group,the mRNA and protein expressions of VEGF,VEGF-C were increased in PD 2-week group (P ＜ 0.05); Compared with PD 2-week group,the expressions were increased in PD 4-week group (P ＜ 0.05).(4) Compared with normal group,the expressions of TTP protein was decreased in uremia group and PD 2-week group (P ＜ 0.05).Compared with uremia and PD2-week group,the expressions of TTP protein was significantly decreased in PD4-week group (P ＜ 0.05).Conclusions High glucose peritoneal dialysis fluid and uremic circumstance result in the expression changes of TTP and VEGF family in a PD time-dependent manner.High glucose peritoneal dialysis liquid gives rise to angiogenesis and lymphangiogenesis,both of which lead to UFF.