黄芪对高糖腹透液诱导大鼠腹膜间皮细胞EMT中TGF-β1/Smads信号通路的影响  

Effect of Radix Astragali on TGF-β1/Smads Signaling Pathway in Epithelial-mesenchymal Transition of rat Peritoneal Mesothelial Cells Induced by High-glucose Peritoneal Dialysate

作　　者：李正红[1] 盛梅笑[2] 张旭[3] 张露[2] 朱长乐[2] 杨劲松[1] 

机构地区：[1]南京中医药大学第一临床医学院,江苏南京210023 [2]南京中医药大学附属医院,江苏南京210029 [3]南京中医药大学研究生院,江苏南京210023

出　　处：《南京中医药大学学报》2013年第3期227-232,261,共7页Journal of Nanjing University of Traditional Chinese Medicine

基　　金：江苏省自然科学基金(BK2011869);江苏省中医药领军人才项目(LJ200904);江苏省高校优势学科建设工程资助项目(PAPD)

摘　　要：目的观察黄芪对高糖腹透液诱导大鼠腹膜间皮细胞EMT中TGF-β1/Smads信号蛋白的影响,探讨黄芪对腹膜纤维化的阻抑作用与机制。方法实验分为5组,每组10只。阴性对照组:每日腹腔注射生理盐水20mL/只,共35d;模型组:每日腹腔注射4.25%葡萄糖腹透液20mL/只,共35d;黄芪低、中、高剂量组:于造模第16天开始每日腹腔注射黄芪腹透液(腹透液中加入黄芪注射液,分别含黄芪生药终浓度10、20、40mg/mL)20mL/只,共20d。HE染色观察腹膜病理改变;快速免疫组化法检测腹膜组织TGF-β1、CollagenⅠ、CollagenⅢ、FN、E-cadherin、α-SMA的表达;Western blot检测腹膜组织TGF-β1、E-cadherin、α-SMA、Smad2/3、p-smad2/3、Smad7蛋白的表达;RT-PCR检测TGF-β1、E-cadherin、α-SMA、Smad7mRNA表达。结果①模型组PMCs呈圆形或柱形,有脱落,间皮下基质增生,大量成纤维样细胞及单核、巨噬细胞浸润,纤维素样物质沉积,腹膜组织TGF-β1、CollagenⅠ、CollagenⅢ、FN表达上调,黄芪各干预组上述变化有改善,以高剂量组作用较为明显;②模型组EMT标记蛋白α-SMA表达上调,E-cadherin表达下调,高、中剂量黄芪组能一定程度地逆转腹膜组织E-cadherin表达下降与α-SMA高表达;③模型组Samds信号蛋白p-smad2/3、Smad7表达上调,黄芪可不同程度地下调p-smad2/3高表达,增强Samd7的表达,高剂量组作用较为显著。结论高糖腹透析液可促使PMCs EMT的发生,黄芪很可能通过抑制TGF-β1,并作用于Smads信号转导通路正、负反馈环路中的Smad2/3、Smad7关键信号蛋白,阻抑PMCs EMT的发生。OBJECTIVE To observe the effect of Radio： Astragali on TGF-β1/Smads signaling pathway in epithelial-mesen- chymal transition（EMT） of rat peritoneal mesothelial cells induced by high-glucose peritoneal dialysis and explore the inhibitory influence and mechanism of Radix Astragali on peritoneal fibrosis. METHODS They were divided into 2 groups, 10 rats in each. The contrast group＇s rats were intraperitoneal injected 35days normal saline 20 mL/rat. The model group＇s rats were in- traperitoneal injected 35days lcodextrin 20 mL/rats. The low/middle and high doses astragalus mongholicus groups： After 16 days modeling, the rats were intraperitoneal injected 20 days astragalus mongholicus extraneal（put astragalus mongholicus in- jections into the externeal, and the concentation of crude drug is 10 mg/mL, 20 mg/mL and 40 mg/mL） 20 mL/rats. The peri- toneal pathological changes were observed through HE staining; the expressions of abdominal membrane TGF-β1, Collagen Ⅰ, Collagen Ⅲ, FN, E-cadherin, α-SMA wece examined through rapid immunohistochemical method; the expressions of proteins of peritoneal tissues TGF-β1, E-cadherin, α-SMA, Smad2/3, p-smad2/3, of Smad7 were detected through Westernblot; the expressions of TGF-β1, E-eadherin, α-SMA, SmadTmRNA were detected through RT-PCR. RESULTS ①PMCs in model group were round or oval with cells falling off, subcutaneous matrix proliferating, a large number of fibroblast-like cells and monocytes and macrophage infiltrating, fibrinoid material depositing and the expression of peritoneal tissues TGF-β1, Collagen Ⅰ, Collagen Ⅲ and FN increasing. The astragalus intervention groups showed improvement in the above conditions with the high-dose group the most obvious. ②The expression ofα -SMA in model group labeled by EMT showed an up-regulation whilethat of E-cadherin low-regulation. The middle and high doses of Radix Astragali groups could reverse the decrease in expres- sion of E-cadherin in peritoneal tissues and high expression of a-SMA to some e

关 键 词：黄芪注射液 腹膜纤维化 PMCs EMT TGF-β1 SMADS信号通路 

分 类 号：R285.5[医药卫生—中药学]