摘要
目的:检测转化生长因子β1在腹膜透析大鼠腹膜内表达,并探讨其在腹膜纤维化中的意义。方法:实验于2005-06/2006-03在中南大学湘雅二医院肾内科实验室完成。①实验材料:雄性SD大鼠,体质量180～240g,由中南大学湘雅二医院动物实验中心提供。②实验方法:将28只大鼠按随机数字表随机分为4组,每组7只。正常对照组不予任何干预;生理盐水组腹腔注射20mL生理盐水;低糖透析液组腹腔注射20mL1.5%葡萄糖透析液;高糖透析液组腹腔注射20mL4.25%葡萄糖透析液,均为1次/d。4周后,向大鼠腹腔注射4.25%葡萄糖腹膜透析液20mL,4h后于大鼠右下腹缓慢插入带有多个侧孔的10号静脉留置针,缓慢低位引流腹透液,量取引流液。③实验评估:取大鼠壁层腹膜组织,以苏木素-伊红染色,镜下测量腹膜厚度,采用免疫组织化学方法检测大鼠腹膜中转化生长因子β1及纤连蛋白。结果:28只大鼠均进入结果分析。①高糖透析液组、低糖透析液组超滤量均明显低于正常对照组与生理盐水组,并且高糖透析液组超滤量明显少于低糖透析液组(P均<0.05)。②高糖透析液组腹膜明显增厚,表面粗糙,间皮细胞肿胀,脱落,间皮下有大量血管生成以及胶原纤维沉积,还可见单核细胞等炎症细胞浸润,与其他组比较,腹膜厚度明显增加(P<0.05)。③高糖透析液组转化生长因子β1、纤连蛋白表达量均明显高于其他组;低糖透析液组转化生长因子β1、纤连蛋白表达量均明显高于正常对照组与生理盐水组(P<0.05)。④大鼠腹膜组织转化生长因子β1蛋白与纤连蛋白表达量、腹膜厚度之间呈明显的正相关(r=0.86,0.83,P<0.05)。结论:葡萄糖透析液可诱导腹膜组织转化生长因子β1明显上调,腹膜转化生长因子β1高表达与腹膜透析腹膜纤维化密切相关。
AIM： To investigate the expression of transforming growth factor β1 （TGF-β1） in the peritoneum of peritoneal dialysis rat, and explore the potential role of TGF-β1 in the progress of peritoneal fibrosis. METHODS： The experiment was carried out in the laboratory of Nephrology Department at the Second Xiangya Hospital of Central South University from June 2005 to March 2006.①SD male rats weighed 180-240 g were offered by the Animal Experimental Center at the Second Xiangya Hospital of Central South University. ②Twenty-eight rats were divided into four groups at random （n =7）. Control group received no treatment; Physiological saline group received intraperitoneal injection of 20 mL saline; Low glucose （LG） group was treated with 20 mL glucose dialysate （1.5%） by intraperitoneal injection, while high glucose （HG） group with equal dialysate （4.25%）. All the injections were given once a day for 4 weeks. Then the rats were intraperitoneally injected with 20 mL HG dialysate, 4 hours later, No. 10 venous retaining needle with many lateral apertures was inserted gradually into right lower quadrant of rats to drain and obtain dialysate. ③The visceral peritoneum tissues of rats were harvested and stained by hematoxylin-eosin. The thickness of peritoneal membrane in peritoneum was measured. TGF-β1 and fibronectin （FN） were detected by immunohistochemistry. RESULTS： All of 28 rats were involved in the result analysis.①Ultrafiltration volume in LG and HG groups were obviously decreased, and HG group was significantly lower than LG group （P 〈 0.05）.②The peritoneal thickness in HG group increased significantly compared with other groups （P 〈 0.05）, the surface was tough, mesothelial cells swelled and ablated. There ware generous angiogenesis and collagen fiber deposition found under mesothelial cells, with inflammatory cells infiltrated. ③The expressions of TGF-β1 and FN up-regulated significantly in HG and LG groups, compared with control group and physiological saline group （P 〈 0.05）.④The expression of TGF-β1 protein in peritoneal tissues was positively correlated with FN protein＇s expression and the thickness of peritoneum （r =0.86, 0.83, P 〈 0.05）. CONCLUSION： Glucose-based dialysate can up-regulate the expression of TGF-β1 remarkably and contribute to peritoneal fibrosis.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2007年第36期7157-7160,共4页
Journal of Clinical Rehabilitative Tissue Engineering Research
