加味补阳还五汤对术后腹腔粘连TGF-β1/Smad3通路和腹膜间皮细胞上皮-间充质转化的影响  

Effect of Modified Buyang Huanwu Decoction(加味补阳还五汤)on TGF-β1/Smad3 Signal Path and Epithelial-Mesenchymal Transition in Postoperative Abdominal Adhesions

作　　者：郑敏麟[1] 范文江 王亚楠 詹倩倩 ZHENG Minlin;FAN Wenjiang;WANG Yanan;ZHAN Qianqian(School of Integnative Medicine,Fujian University of Traditional Chinese Medieine,F uzhou 350122,Fujian.China;Research Institute of Integnative Medicine,Fujian University of Tradtional Chinese Medicine,Fuhou 350122,Fujian,China)

机构地区：[1]福建中医药大学中西医结合学院,福建福州350122 [2]福建中医药大学中西医结合研究院,福建福州350122

出　　处：《中华中医药学刊》2024年第5期20-27,I0007-I0013,共15页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(82274516);福建省自然科学基金面上项目(2022J01842);福建中医药大学校管科研课题-重点项目(自然科学)(X2021003-重点)。

摘　　要：目的探讨加味补阳还五汤对术后腹腔粘连转化生长因子β1(transforming growth factor-beta1,TGF-β1)/Smad3通路和腹膜间皮细胞的上皮-间充质转化的影响。方法将108只雄性SD大鼠随机分为正常组、模型组、透明质酸钠组、加味补阳还五汤组。模型组、透明质酸钠组及加味补阳还五汤组进行术后腹腔粘连模型造模,透明质酸钠为阳性对照组,在手术造模后,一次性腹腔喷洒1%的透明质酸钠凝胶,0.5mL/kg。加味补阳还五汤组术后第2天开始灌胃,灌胃剂量为19.5g/(kg·d)。分别于术后7、14、28d分批将动物处死,每次每组处死9只大鼠。在损伤部位取材:应用免疫组织化学法观察粘连部位的纤维化通路相关蛋白TGF-β1、Smad3、Smad7的表达情况;以及其下游胶原沉积相关蛋白Ⅰ型胶原(CollagenⅠ,Col-Ⅰ)、Ⅲ型胶原(CollagenⅢ,Col-Ⅲ)的表达情况和上皮-间充质转化(Epithelialmesenchy-maltransition,EMT)相关的蛋白E-钙黏蛋白(E-Cadherin)、α-平滑肌肌动蛋白(alpha-smoothmuscleactin,α-SMA)的表达情况。结果①不同组别间,术后7、14、28d3个时间节点,TGF-β1/Smad3纤维化通路相关蛋白表达情况:与正常组比较,模型组TGF-β1蛋白表达增多(均为P<0.01);与模型组比较,透明质酸钠组和加味补阳还五汤组TGF-β1蛋白表达减少(均为P<0.05)。与正常组比较,模型组Smad3蛋白表达增多(均为P<0.05);与模型组比较,透明质酸钠组和加味补阳还五汤组Smad3蛋白表达减少(P<0.05)。与正常组比较,模型组Smad7蛋白表达减少(均为P<0.01);与模型组比较,透明质酸钠组和加味补阳还五汤组Smad7蛋白表达增多(均为P<0.05)。②胶原沉积情况:术后7、14、28d3个时间节点,Col-Ⅰ蛋白表达存在差异:与正常组比较,模型组Col-Ⅰ蛋白表达增多(均P<0.01);与模型组比较,透明质酸钠组和加味补阳还五汤组Col-Ⅰ蛋白表达减少(均P<0.01)。与正常组比较,模型组Col-Ⅲ蛋白表达增多(均P<0.01Objective To explore the possible mechanism of Modified Buyang Huanwu Decoction(加味补阳还五汤)in the treatment of postoperative abdominal adhesions related to transforming growth factor-β1(TGF-β1)/Smad3 pathway and epithelial-mesenchymal transition in peritoneal mesothelial cells.Methods One hundred and eight male SD rats were randomly divided into normal group,model group,sodium hyaluronate group and Modified Buyang Huanwu Decoction group.The model group,sodium hyaluronate group and Modified Buyang Huanwu Decoction group established the model of postoperative abdominal adhesion.The sodium hyaluronate group was as the positive control group.After surgical modeling,one-time intraperitoneal spraying of 1%sodium hyaluronate gel,0.5 mL/kg.The Modified Buyang Huanwu Decoction group was given the decoction by gavage with the dose of 19.5 g(kg·d)on the second day after operation.The animals were sacrificed 7,14 and 28 days after the operation,9 animals per group were killed each time,and the samples were taken out from the injury site.The immunohistochemistry method was used to observe the expressions of injury site fibrosis pathway-related proteins(TGF-β1,Smad3,Smad7),the expressions of downstream collagen deposition related proteins(Col-Ⅰand Col-Ⅲ)and the epithelial-mesenchymal transition-related proteins[E-cadherin andα-smooth muscle actin(α-SMA)]were all observed by immunohistochemistry.Results①Among different groups,there were 3 time nodes on the 7th,14th and 28th day after operation.TGF-β1/Smad3 fibrosis pathway-related protein expression:Compared with that of the normal group,the content of TGF-β1 protein in the model group was increased(all P<0.01).Compared with that of the model group,the contents of TGF-β1 protein in the sodium hyaluronate group and the Modified Buyang Huanwu Decoction group were decreased(all P<0.05).Compared with that of the normal group,the expression of Smad3 protein in the model group was increased(all P<0.05).Compared with that of the model group,the expressions of

关 键 词：术后腹腔粘连 加味补阳还五汤 纤维化 上皮-间充质转化 TGF-β1/Smad3 

分 类 号：R289.5[医药卫生—方剂学]