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

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。     

血管内皮生长因子和内皮抑素在人腹膜组织的表达及其与腹膜毛细血管新生的关系

目的 检测血管内皮生长因子（VEGF）和血管内皮抑素（ES）在人腹膜组织表达,探讨两者与腹膜血管新生之间的关系。 方法 取健康对照者、尿毒症非透析患者以及腹透患者的腹膜标本,用反转录聚合酶链反应（RT-PCR）检测VEGF和ES mRNA的表达;组织免疫组化染色检测VEGF和ES蛋白质水平的表达;CD34染色计数腹膜组织毛细血管密度（MVD）。 结果 各组腹膜均有VEGF及ES表达;健康对照组、尿毒症非透析组、腹透组VEGF mRNA的相对表达量依次为0.47±0.01、0.62±0.02、0.74±0.02。VEGF免疫组化染色阳性区平均灰度值依次为95.673±2.01、117.126±2.07、140.184±2.25。ES免疫组化染色阳性区平均灰度值依次为94.902±2.38、113.380±2.33、145.489±3.05。尿毒症非透析组、腹透组VEGF mRNA和蛋白表达水平及ES蛋白表达水平表达均高于健康对照组,且腹透组升高更为明显,差异均具有统计学意义（均P < 0.05）。3组ES在mRNA水平表达量依次为0.42±0.02、0.43±0.03、0.43±0.02,各组表达差异无统计学意义（P > 0.05）。3组腹膜MVD依次为3.05±0.45、5.98±0.47、9.62±0.49,尿毒症非透析组、腹透组均高于健康对照组,且腹透组增高更为明显,差异均具有统计学意义（均P < 0.05）。 结论 腹膜透析患者腹膜组织VEGF mRNA和蛋白表达水平升高,ES蛋白表达水平也升高,这可能在长期透析所致腹膜组织新生毛细血管形成过程中发挥一定作用。     

血管生成素2与腹膜透析时腹膜血管新生的关系

目的 观察血管生成素2（Angpt-2）和腹膜透析（腹透）时腹膜血管新生的关系。 方法 5/6肾切除制作尿毒症大鼠模型,成模后在腹腔内植入腹透管,根据大鼠体质量每天经腹透管注入定量腹透液（4.25%,Dineal）。按腹透时间分为未腹透组、腹透10 d组、28 d组及56 d组。假手术非尿毒症非透析大鼠为对照组。大网膜抗CD31免疫组化染色后作血管计数,观察腹膜血管新生。用实时定量PCR和Western 印迹分别检测腹膜Angpt-2和血管内皮生长因子（VEGF）表达量变化,同时检测腹膜血管数和Angpt-2、VEGF表达量的关系。 结果 未腹透组、腹透10 d、28 d及56 d组腹膜血管数显著高于对照组[（5±3）、（10±5）、（17±5）及（19±4）比（1±1） 个/HP,均P < 0.05]。腹透28 d组腹膜血管数显著高于腹透10 d组（P < 0.05）,但与56 d组差异无统计学意义。未腹透组或腹透各组腹膜Angpt-2和VEGF表达显著高于对照组（均P < 0.01）,而Angpt-2和VEGF表达量并未随透析时间延长而增加。Angpt-2和VEGF表达量和腹膜血管数呈正相关（r = 0.7756,P < 0.01;r = 0.5223,P < 0.05）。 结论 尿毒症状态和腹透促进腹膜血管新生。腹膜Angpt-2表达增加和腹膜血管新生呈正相关。Angpt-2将可能成为治疗腹膜血管新生的另一靶点     

正常大鼠腹膜透析模型中腹膜血管新生与HIF-1α及VEGF的表达研究

目的：探讨正常大鼠腹膜透析模型中HIF-1α及VEGF对腹膜血管新生。方法：建立正常大鼠腹膜透析模型,随机分组为正常组、假手术组、1.5%腹透液组及4.25%腹透液组,采用RT—PCR、免疫组化技术研究HIF-1α及VEGF在腹膜组织中的表达,同时观察腹膜血管新生改变。结果：在两组腹透组中HIF-1α及VEGF基因表达明显增高,与正常组及假手术组相比较有差异,并且腹膜血管新生改变。结论：HIF-1α及VEGF在慢性肾衰竭腹膜透析模型腹膜组织中存在基因表达,可能通过促进腹膜血管新生,参与腹膜透析超滤改变。     

内皮抑素在大鼠腹膜的表达及其与腹膜血管新生的关系

Objective To investigate the expression of endostatin (ES) in rat peritoneum and its association with peritoneal neoangiogensis. Methods Thirty-two male SD rats were randomly divided into 4 groups: normal control rats (C group), renal failure without PD rats (non-PD group), rats dialyzed with 1.5% PD solution (1.5% PD group) and 4.25% PD solution (4.25% PD group). After regular PD for 28 days, mRNA and protein expression of ES in peritoneal tissues of each group were detected by RT-PCR and immunohistochemistry respectively. Microvessel density (MVD) of peritoneal tissue was assessed using immunohistochemistry with CD34 monoclonal antibody. Results ES mRNA was expressed in each group, 0.47±0.05 in C group, 0.45±0.04 in non-PD group, 0.46±0.04 in 1.5%PD group, 0.47±0.03 in 4.25%PD group, and no significant differences were found among groups. Score of ES protein expression was O in C group, 2 in non-PD group, 4 in 1.5%PD group, and 9 in 4.25%PD group. MVD was 3.13±1.13 in C group, 5.13±1.14 in non-PD group, 9.00±1.51 in 1.5%PD group, 10.75±1.83 in 4.25%PD group, and significant differences were found among groups. Conclusion Uremia circumstance and non-physiological compatibility peritoneal dialysate can increase ES protein expression and MVD, which may participate in and have effects on the course of peritoneal neoangiogensis.     

内皮抑素在大鼠腹膜的表达及其与腹膜血管新生的关系

Objective To investigate the expression of endostatin (ES) in rat peritoneum and its association with peritoneal neoangiogensis. Methods Thirty-two male SD rats were randomly divided into 4 groups: normal control rats (C group), renal failure without PD rats (non-PD group), rats dialyzed with 1.5% PD solution (1.5% PD group) and 4.25% PD solution (4.25% PD group). After regular PD for 28 days, mRNA and protein expression of ES in peritoneal tissues of each group were detected by RT-PCR and immunohistochemistry respectively. Microvessel density (MVD) of peritoneal tissue was assessed using immunohistochemistry with CD34 monoclonal antibody. Results ES mRNA was expressed in each group, 0.47±0.05 in C group, 0.45±0.04 in non-PD group, 0.46±0.04 in 1.5%PD group, 0.47±0.03 in 4.25%PD group, and no significant differences were found among groups. Score of ES protein expression was O in C group, 2 in non-PD group, 4 in 1.5%PD group, and 9 in 4.25%PD group. MVD was 3.13±1.13 in C group, 5.13±1.14 in non-PD group, 9.00±1.51 in 1.5%PD group, 10.75±1.83 in 4.25%PD group, and significant differences were found among groups. Conclusion Uremia circumstance and non-physiological compatibility peritoneal dialysate can increase ES protein expression and MVD, which may participate in and have effects on the course of peritoneal neoangiogensis.     

内皮抑素在大鼠腹膜的表达及其与腹膜血管新生的关系

Objective To investigate the expression of endostatin (ES) in rat peritoneum and its association with peritoneal neoangiogensis. Methods Thirty-two male SD rats were randomly divided into 4 groups: normal control rats (C group), renal failure without PD rats (non-PD group), rats dialyzed with 1.5% PD solution (1.5% PD group) and 4.25% PD solution (4.25% PD group). After regular PD for 28 days, mRNA and protein expression of ES in peritoneal tissues of each group were detected by RT-PCR and immunohistochemistry respectively. Microvessel density (MVD) of peritoneal tissue was assessed using immunohistochemistry with CD34 monoclonal antibody. Results ES mRNA was expressed in each group, 0.47±0.05 in C group, 0.45±0.04 in non-PD group, 0.46±0.04 in 1.5%PD group, 0.47±0.03 in 4.25%PD group, and no significant differences were found among groups. Score of ES protein expression was O in C group, 2 in non-PD group, 4 in 1.5%PD group, and 9 in 4.25%PD group. MVD was 3.13±1.13 in C group, 5.13±1.14 in non-PD group, 9.00±1.51 in 1.5%PD group, 10.75±1.83 in 4.25%PD group, and significant differences were found among groups. Conclusion Uremia circumstance and non-physiological compatibility peritoneal dialysate can increase ES protein expression and MVD, which may participate in and have effects on the course of peritoneal neoangiogensis.     

内皮抑素在大鼠腹膜的表达及其与腹膜血管新生的关系

Objective To investigate the expression of endostatin (ES) in rat peritoneum and its association with peritoneal neoangiogensis. Methods Thirty-two male SD rats were randomly divided into 4 groups: normal control rats (C group), renal failure without PD rats (non-PD group), rats dialyzed with 1.5% PD solution (1.5% PD group) and 4.25% PD solution (4.25% PD group). After regular PD for 28 days, mRNA and protein expression of ES in peritoneal tissues of each group were detected by RT-PCR and immunohistochemistry respectively. Microvessel density (MVD) of peritoneal tissue was assessed using immunohistochemistry with CD34 monoclonal antibody. Results ES mRNA was expressed in each group, 0.47±0.05 in C group, 0.45±0.04 in non-PD group, 0.46±0.04 in 1.5%PD group, 0.47±0.03 in 4.25%PD group, and no significant differences were found among groups. Score of ES protein expression was O in C group, 2 in non-PD group, 4 in 1.5%PD group, and 9 in 4.25%PD group. MVD was 3.13±1.13 in C group, 5.13±1.14 in non-PD group, 9.00±1.51 in 1.5%PD group, 10.75±1.83 in 4.25%PD group, and significant differences were found among groups. Conclusion Uremia circumstance and non-physiological compatibility peritoneal dialysate can increase ES protein expression and MVD, which may participate in and have effects on the course of peritoneal neoangiogensis.     

内皮抑素在大鼠腹膜的表达及其与腹膜血管新生的关系

Objective To investigate the expression of endostatin (ES) in rat peritoneum and its association with peritoneal neoangiogensis. Methods Thirty-two male SD rats were randomly divided into 4 groups: normal control rats (C group), renal failure without PD rats (non-PD group), rats dialyzed with 1.5% PD solution (1.5% PD group) and 4.25% PD solution (4.25% PD group). After regular PD for 28 days, mRNA and protein expression of ES in peritoneal tissues of each group were detected by RT-PCR and immunohistochemistry respectively. Microvessel density (MVD) of peritoneal tissue was assessed using immunohistochemistry with CD34 monoclonal antibody. Results ES mRNA was expressed in each group, 0.47±0.05 in C group, 0.45±0.04 in non-PD group, 0.46±0.04 in 1.5%PD group, 0.47±0.03 in 4.25%PD group, and no significant differences were found among groups. Score of ES protein expression was O in C group, 2 in non-PD group, 4 in 1.5%PD group, and 9 in 4.25%PD group. MVD was 3.13±1.13 in C group, 5.13±1.14 in non-PD group, 9.00±1.51 in 1.5%PD group, 10.75±1.83 in 4.25%PD group, and significant differences were found among groups. Conclusion Uremia circumstance and non-physiological compatibility peritoneal dialysate can increase ES protein expression and MVD, which may participate in and have effects on the course of peritoneal neoangiogensis.     

内皮抑素在大鼠腹膜的表达及其与腹膜血管新生的关系

Objective To investigate the expression of endostatin (ES) in rat peritoneum and its association with peritoneal neoangiogensis. Methods Thirty-two male SD rats were randomly divided into 4 groups: normal control rats (C group), renal failure without PD rats (non-PD group), rats dialyzed with 1.5% PD solution (1.5% PD group) and 4.25% PD solution (4.25% PD group). After regular PD for 28 days, mRNA and protein expression of ES in peritoneal tissues of each group were detected by RT-PCR and immunohistochemistry respectively. Microvessel density (MVD) of peritoneal tissue was assessed using immunohistochemistry with CD34 monoclonal antibody. Results ES mRNA was expressed in each group, 0.47±0.05 in C group, 0.45±0.04 in non-PD group, 0.46±0.04 in 1.5%PD group, 0.47±0.03 in 4.25%PD group, and no significant differences were found among groups. Score of ES protein expression was O in C group, 2 in non-PD group, 4 in 1.5%PD group, and 9 in 4.25%PD group. MVD was 3.13±1.13 in C group, 5.13±1.14 in non-PD group, 9.00±1.51 in 1.5%PD group, 10.75±1.83 in 4.25%PD group, and significant differences were found among groups. Conclusion Uremia circumstance and non-physiological compatibility peritoneal dialysate can increase ES protein expression and MVD, which may participate in and have effects on the course of peritoneal neoangiogensis.     

Effect of peritoneal dialysis on expression of vascular endothelial growth factor, basic fibroblast growth factor and endostatin of the peritoneum in peritoneal dialysis patients

Aim: The aim of this study is to investigate the expression of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF) and endostatin (ES) in human peritoneum and investigate the relationship between them and peritoneum neoangiogensis in the patients with uraemia and peritoneal dialysis (PD). Methods: Peritoneal biopsies were obtained from normal subjects (n = 8), uraemic predialysis patients (n = 12) and PD patients (n = 10). The mRNA expression of VEGF, bFGF and ES in peritoneal tissues were measured through real‐time polymerase chain reaction. The protein expression of VEGF, bFGF and ES in peritoneal tissues were determined through western blot. Microvessel density (MVD) of peritoneal tissue was assessed using immunohistochemistry with CD34 monoclonal antibody. Results: The mRNA and protein of VEGF, bFGF and ES were expressed in all peritoneal samples. Compared with the normal control group, the mRNA and protein expression of VEGF and bFGF in peritoneal tissues were all significantly upregulated in the uraemic predialysis and PD group (all P < 0.05). Compared with the normal control group, the protein expression of ES were significantly upregulated in the uraemic predialysis and PD group (all (P < 0.05), but the mRNA expression of ES did not have obvious differences in the uraemic predialysis and PD group as compared to the normal control group (P > 0.05). MVD of peritoneal tissue were increased in the uraemic predialysis and PD group compared with the normal group (all P < 0.05). A significant positive correlation was found between VEGF mRNA expression and MVD, bFGF mRNA expression and MVD. Conclusion: The mRNA expression of VEGF and bFGF, the protein expression of VEGF, bFGF, and ES and microvessel density (MVD) are increased both in the uraemic predialysis and PD patients. These results show that uraemia circumstances and non‐physiological compatibility of peritoneal dialysis solution might increase VEGF, bFGF and ES expression and MVD, which might participate in the increment of the peritoneum neoangiogensis and ultrafiltration failure in PD patients.     

可溶性Tie2融合蛋白对尿毒症腹膜透析大鼠血管新生的影响

目的 观察可溶性Tie2融合蛋白(sTie2/Fc)对尿毒症腹膜透析大鼠腹膜血管新生的影响.方法 48只雄性SD大鼠,按随机数字表法分为以下6组:正常对照组、假手术组、尿毒症非腹透组、4.25％腹透组、sTie2/Fc 2.5 μg/kg干预组、sTie2/Fc 5.0 μg/kg干预组.腹透组按照4.25％腹透液30...     

Effects of cyclooxygenase - 2 inhibitor on peritoneal function and angiogenesis in uremic peritoneal dialysis rats

Objective To investigate the effects of the cyclooxygenase-2 (COX-2) inhibitor (celecoxib) on angiogenesis and peritoneal function of uremic peritoneal dialysis rats. Methods Forty - eight male SD rats were selected, and they were randomly divided into five groups: normal control group(n=8), sham operation group(n=8), uremia group(5/6 nephrectomy, n=8), PD group [4.25% PD solution, 2 weeks PD model(n=8) and 4 weeks PD model(n=8)], PD + celecoxib intervention group[treated by celecoxib(20 mg/kg) via oral gavage, n=8].The peritoneum of uremic peritoneal dialysis rats was observed in different dialysis time from peritoneal structures, functions, peritoneal tissue capillary density (microvessel density, MVD) and COX-2, vascular endothelial growth factor (VEGF) expression level, and the impacts of celecoxib on uremic peritoneal dialysis rats peritoneal angiogenesis and peritoneal function were study. Results With the conduct of the peritoneal dialysis, peritoneal thickness increased, the inflammatory cells infiltrated, peritoneal equilibration test (PET) showed that ultrafiltration volume decreased significantly (P＜0.05), the amount of glucose transport rate rised significantly (P＜0.05), but the celecoxib could improve net ultrafiltration volume (P＜0.05), and reduce the glucose transport rate (P＜0.05). The peritoneal tissue MVD and COX - 2, VEGF expression were significantly increased in uremia group and PD group compared with that in the normal control group (all P＜0.05), were significantly lower in PD + Celecoxib intervention group than that in uremia group (P＜0.05). The correlation analysis showed that the level of COX-2 protein expression with MVD, VEGF protein expression was positively correlated (both P＜0.05), the level of VEGF protein expression and MVD was positively correlated (P＜0.05). Conclusions In vivo high glucose dialysate and uremia environmental can stimulate the COX-2 and VEGF expression raised, and the capillaries production increased in peritoneal tissue. Celecoxib can alleviate the change of peritoneal tissue morphology and function in long-term peritoneal dialysis rats. Celecoxib inhibits the peritoneal neovascularization of uremic peritoneal dialysis rats, possibly through inhibition of COX-2 expression to reduce the production of VEGF.     

扶肾颗粒对腹膜透析相关性腹膜纤维化的影响及其作用机制的实验研究

目的:通过对腹膜透析大鼠进行干预,观察扶肾颗粒对腹膜纤维化相关细胞因子的影响及其作用机制探讨。方法:SPF级健康雄性SD大鼠75只,体重180～200g,适应性饲养1周后,依大鼠体重分层,随机分为:正常对照组(A组,n=15),无任何处理,腹腔注射生理盐水,100ml.kg-1.d-1,连续4周;肾衰5/6切除+1.5%PD组(B组,n=15);肾衰5/6切除+1.5%PD+扶肾颗粒组(C组,n=15);肾衰5/6切除+4.25%PD组(D组,n=15);肾衰5/6切除+4.25%PD+扶肾颗粒组(E组,n=15),除A组外,各组均制成肾衰模型,其中B、C组予腹腔注射1.5%LPDS,100ml.kg-1.d-1,连续4周;D、E组予腹腔注射4.25%LPDS,100ml.kg-1.d-1,连续4周,自透析开始,对C组和E组予灌服扶肾颗粒,以200gSD大鼠计算,灌胃剂量1.8ml/d。其余各组予灌服2ml生理盐水,共灌服4周。观察各组实验动物的大体状态、体重变化、腹膜滤过功能及血清纤维化调控因子表达变化。结果:在治疗4周后,除正常对照组(A组)外,其余各组均体现为负超滤,但是,同浓度透析治疗组中,应用扶肾颗粒干预组其超滤情况明显优于未应用扶肾颗粒组(P<0.05),其作用显著,同时,葡萄糖转运量方面亦体现为相似结果。与正常对照组比较(A组),其余各模型组相关促纤维化因子表达水平均明显升高(P<0.01)。TGF-β1方面,D组的表达水平明显较其他各组升高(P<0.05)。CTGF及IL-6方面,结果与TGF-β1相似。VEGF方面,E组较D组明显降低(P<0.05)。随着透析治疗的进行,HGF与BMP-7表达水平均反应性升高,各模型组二者水平均较正常对照组明显升高(P<0.01)。HGF方面,C组较B组明显升高(P<0.01)。BMP-7方面,E组较D组明显升高(P<0.05)。结论:扶肾颗粒可减轻腹透大鼠的肾功能损伤,保护残余肾功能,改善腹透大鼠的超滤量及葡萄糖转运量,抑制促纤维化因子TGF-β1、CTGF、IL-6、CTGF等表达,调高抗纤维化因子HGF、BMP-7的表达,进而起到抑制腹膜透析相关腹膜纤维化的发生,改善腹膜透析效能。     

Effect of the JAK2/STAT3 signaling pathway on epithelial mesenchymal transition of the peritoneum in uremic peritoneal dialysis rats

Objective To investigate whether the JAK2/STAT3 signaling pathway is involved in the epithelial-mesenchymal transition (EMT) of peritoneal mesothelial cells in uremic peritoneal dialysis (PD) rats. Methods A total of 48 male Sprague-Dawley (SD) rats were randomly separated into six groups: normal control group (NC group, n=8), sham group (n=8), uremic group (n=8), PD group (n=8), S3I-201 control group (n=8) and S3I-201 group (n=8). Uremic model generated by 5/6 nephrectomy surgery in rats was established. The rats of PD group, S3I-201 control group and S3I-201 group received daily infusion of 4.25% glucose-based peritoneal dialysate fluid (3 ml/100 g) from PD catheters for 28 days. Rats of S3I-201 group were injected with STAT3 inhibitor S3I-201 (2.5 mg/kg) solution from the catheters every other day; the same dose of the solvent of S3I-201 was simultaneously given to S3I-201 control group rats. After PD for 28 days, peritoneal function, pathologic changes, and microvessel density (MVD) were evaluated. Creatinine, urea nitrogen and interleukin-6 (IL-6) concentration in blood and dialysate, and protein and mRNA levels of phospho-JAK2 (p-JAK2), phospho-STAT3 (p-STAT3), E-cadherin, alpha-smooth muscle actin (α-SMA) and vascular endothelial growth factor (VEGF) in peritoneum were determined. Results Uremia and peritoneal dialysate could aggravate the peritoneal function and elevate peritoneal thickness and MVD. They could also increased the concentration of IL-6 in blood and dialysate and the expression levels of α-SMA, VEGF, p-JAK2 and p-STAT3 in peritoneum, while lowering E-cadherin expression in peritoneum. These manifestations were even more remarkable in PD group compared to those in uremic group. There was no statistical difference between the S3I-201 control group and the PD group as regards all the index (all P＞0.05). Compared with the S3I-201 control group, the rats treated with S3I-201 showed better peritoneal function. S3I-201 could reduce peritoneal thickness (P＜0.05), MVD (P＜0.05), the concentration of IL-6 in blood and dialysate, the mRNA and protein expression of α-SMA, VEGF, p-JAK2 and p-STAT3 (all P＜0.05), while enhance the mRNA and protein expression of E-cadherin (all P＜0.05). Conclusions After STAT3 is inhibited, the peritoneal thickness, MVD and IL-6 concentration in PD rats are decreased, and EMT is also inhibited, while peritoneal function is improved. The JAK2/STAT3 signaling pathway may thus be involved in the process of EMT of peritoneum in uremic peritoneal dialysis rats by regulating the expression of IL-6.     

Effect of fluvastatin on transforming growth factor beta 1 expression in peritoneal dialysis rats model

ObjectiveTo investigate the effect of fluvastatin on TGF-β1 expression in a rat model of peritoneal dialysis(PD). MethodsA rat model of PD was built by intraperitoneal injection of 2.5% or 4.25% peritoneal dialysate. SD rats were randomly divided into 7 groups: (1) Normal control group; (2)Saline control group: saline 100 ml/kg intraperitoneal injection(IP) each day; (3) Fluvastatin treatment group: fluvastatin intragastrically administration 10 mg/kg each day; (4) 2.5% PD group: 2.5% peritoneal dialysate IP 100 ml/kg everyday; (5)4.25% PD group: 4.25% peritoneal dialysate IP 100 ml/kg everyday; (6)2.5% PD plus fluvastatin treatment group: 2.5% peritoneal dialysate IP 100 ml/kg plus fluvastatin 10 mg/kg everyday; (7)4.25% PD plus fluvastatin treatment group: 4.25% peritoneal dialysate IP 100 ml/kg plus fluvastatin 10 mg/kg everyday. The rats were sacrificed at 6 weeks and peritoneal tissues were dissected. The expressions of TGF-β1 and FN were examined by RT-PCR and immunohistochemical analysis. Masson staining was used for histological examination. ResultsMasson staining showed that the peritoneum thickened in 2.5% and 4.25% PD group than in normal control group and saline control group. The fluvastatin treatment ameliorated the thickening of peritoneum induced by PD. RT-PCR and immunohistochemical analysis showed that the mRNA and protein expression of TGF- β1 and FN increased in 2.5% and 4.25% PD group than in normal and saline control group (all P＜0.05). The fluvastatin treatment ameliorated the increased expression of TGF-β1 and FN induced by PD. There was no statistically significant difference among normal control group, saline control group and fluvastatin treatment group in both peritoneal thickness and the expression of TGF-β1 and FN. ConclusionFluvastatin can reduce the increased expressions of TGF -β1 and FN in rat peritoneum and ameliorate the thickening of peritoneum induced by PD.     

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

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.