褪黑素通过调节TGF-β1/Smad通路抗腹膜透析相关性腹膜纤维化  

Effect of melatonin on anti peritoneal dialysis-related peritoneal fibrosis by regulating the TG-β1/Smad pathway

作　　者：史伟文 熊燕移 李敏芝 吴永贵[2] 卓华钦 齐向明[2] 赵珉[2] 黄丹丹[2] 

机构地区：[1]东莞市第三人民医院肾内科,广东东莞523326 [2]安徽医科大学第一附属医院肾内科,安徽合肥230000

出　　处：《中国临床研究》2017年第5期626-630,共5页Chinese Journal of Clinical Research

基　　金：广东省医学科学技术研究基金(A2014867)

摘　　要：目的探讨褪黑素(N-乙酰-5-甲氧色氨酸,MT)通过调节TGF-β1/Smad通路抗腹膜透析相关性腹膜纤维化的机制。方法将选取的60只大鼠随机分为正常对照组、模型对照组、低剂量药物组、中剂量药物组、高剂量药物组,每组12只。除正常对照组外,其余各组大鼠均采用腹腔注射4.25%的葡萄糖腹膜透析液制作腹膜纤维化模型。造模后,低剂量组、中剂量组、高剂量组分别给予MT 0.125、0.5、2.0 mg·kg-1·d-1灌胃;正常对照组及模型对照组仅给予等体积生理盐水灌胃。适宜条件下培育4周后,处死大鼠并取适量腹膜、肠道组织及血液标本。腹膜、肠道组织行HE及Masson三色染色,观察腹膜和肠道黏膜病理组织学及壁层腹膜厚度变化。免疫组织化学法检测腹膜组织中转化生长因子-β1(TGF-β1)、α平滑肌肌动蛋白(α-SMA)、单核细胞趋化蛋白-1(MCP-1)及肠黏膜Occludin蛋白相对表达量;凝胶电泳迁移分析法(EMSA)方法测定核因子(NF)-κB活性;双抗体夹心ELISA法检测样本中白介素-1(IL-1)、肿瘤坏死因子α(TNF-α)浓度。结果模型对照组的大鼠壁层腹膜厚度、IL-1、TNF-α水平显著高于正常对照组和低、中、高剂量组(P<0.05,P<0.01);高剂量组的大鼠壁层腹膜厚度、IL-1、TNF-α水平显著低于模型对照组和低、中剂量组(P<0.05,P<0.01)。模型对照组的TGF-β1、MCP-1、α-SMA、Occludin蛋白表达水平及核因子(NF)-κB活性显著高于正常对照组和低、中、高剂量组(P<0.05,P<0.01),高剂量组的TGF-β1、MCP-1、α-SMA、Occludin表达水平及NF-κB活性显著低于模型对照组和低、中剂量组(P<0.05,P<0.01),高剂量组的TNF-α、α-SMA表达水平与正常对照组无统计学差异(P均>0.05)。结论高糖腹腔注射可制作腹膜纤维化模型。MT可以适度调节TGF-β1/Smad通路、抑制NF-κB信号的传导,抑制引起腹膜纤维化的重要细胞内信号调节因子TGF-β1、NF-κB,缓解腹膜纤维化。Objective To investigate the mechanism of effect of melatonin （ N-acetyl-5-metyoxytryptamine, MT） on anti peritoneal dialysis-related peritoneal fibrosis via regulating the TG-β1/Smad pathway. Methods Sixty rats were randomly divided into normal control groupmodel control group,low-, middle-,high-dose drug groups（ n = 12 each）. Except normal control group, peritoneal fibrosis model was established by intraperitoneal injection of peritoneal dialysis solution containing 4.25 % glucose in rats of other groups. After modeling,0.125-,0.5-,2.0-mg ·kg - 1 ·d - 1 of MT were respectively given by gavage administration in low-, middle-, high-dose drug groups. In normal control group, equal volume of normal saline was given by intraperitonea] injection. After cultivation for 4 weeks under suitable conditions, the rats were killed, and appropriate amount of peritoneal, intestinal tissues and blood samples were taken. The changes of histopathology of peritoneal and intestinal mucosa and parietal peritoneum thickness were observed after HE and Masson trichrome staining. Immunohistochemical method was used to detect the expressions of transforming growth factor-β1 （ TGF-β1 ） , a-smooth muscle action （a-SMA）,monocyte ehemotaetic protein 1 （MCP-1）in peritoneal tissues and Occludin protein in intestinalmucosa. Electrophoretic mobility shift assay（EMSA） was used to detect nuclear factor-kappa B（NF-KB） activity. Enzyme linked immunosorbent assay （ELISA）-sandwich technique was used to detect the concentrations of interleukin -1 （IL -1 ）, tumor necrosis factor （TNF-α）. Results The thickness of parietal peritoneum, IL-1 and TNF-α levels in rats of model control group were significantly higher than those in rats of normal control group and low-, middle-, high-dose drug groups （P 〈 0.05, P 〈 0. 01 ）which in high-dose drug group were significantly lower than those in model control group and low-, middle-dose drug groups （ P 〈 0.05, P 〈 0. 01 ）. The expression levels of TGF-β1

关 键 词：褪黑素 腹膜纤维化 转化生长因子-Β1 核因子-ΚB 

分 类 号：R692.5[医药卫生—泌尿科学]