加味补阳还五汤抑制腹膜间皮细胞上皮-间充质转换防治术后腹腔粘连的实验研究

Study on the Prevention and Treatment of Postoperative Abdominal Adhesion by Inhibiting Epithelial-Mesenchymal Transition of Peritoneal Mesothelial Cells with Supplemented Buyang Huanwu Decoction

目的探讨加味补阳还五汤治疗术后腹腔粘连(Postoperative abdominal adhesions,PAA)的疗效和机制。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PAA模型造模,透明质酸钠阳性对照组。分别于术后7、14、28天分批处死,在损伤部位进行目视评分,取材并进行苏木素-伊红染色、马松染色并进行粘连评级,扫描电镜观察腹膜间皮细胞的超微结构。应用免疫组织化学法观察粘连部位的上皮-间充质转换(EMT)相关细胞标志蛋白E-cadherin及α-SMA的表达情况。结果①加味补阳还五汤防治PAA的疗效评价:加味补阳还五汤组Diamod目视粘连积分降低(P<0.05),HE染色和Masson染色粘连等级降低(P<0.05)。②腹膜间皮细胞损伤修复情况:扫描电镜显示加味补阳还五汤组视野内能见到铺路状腹膜间皮细胞,表明加味补阳还五汤组能改善PAA盲肠浆膜侧腹膜间皮细胞的损伤。③腹膜间皮细胞EMT相关指标表达情况:加味补阳还五汤组E-Cadherin蛋白表达增多(P<0.05),α-SMA蛋白表达减少(P<0.05)。结论加味补阳还五汤及透明质酸钠均能有效防治PAA,其疗效可能是通过减轻腹膜间皮细胞EMT实现的。

Objective To evaluated the efficacy of Supplemented Buyang Huanwu Decoction in preventing and treating postoperative abdominal adhesions.Methods One hundred and eight male SD rats were randomly divided into ordinary group,mold group,Sodium hyaluronate group and Supplemented Buyang Huanwu Decoction group.The animals were sacrificed 7,14 and 28 days after the operation,Visual scoring was performed,and samples were taken from the injury site for hematoxylin-eosin staining and Masson staining,and scanning electron microscope to observe the morphology of peritoneal mesothelial cells in the modeling site.Immunohistochemical method was used to observe the expression of epithelial mesenchymal transition(EMT)related cell marker proteins E-cadherin andα-SMA in the adhesive site.Results①Efficacy of Supplemented Buyang Huanwu Decoction in the prevention and treatment of postoperative abdominal adhesions:The Supplemented Buyang Huanwu Decoction group can reduce the Diamod visual adhesion score(P<0.05),as well as the adhesion levels of HE staining and Masson staining(P<0.05).②Degree of damage to peritoneal mesothelial cells:Scanning electron microscopy showed that in the field of view of the Supplemented Buyang Huanwu Decoction group,paving like peritoneal mesothelial cells could be seen,indicating that the Supplemented Buyang Huanwu Decoction can improve the damage of peritoneal mesothelial cells on the serosal side of the cecum in PAA.③Expression of Epithelial mesenchymal transition-related indicators:The expression of E-Cadherin protein in the Supplemented BuyangHuanwu Decoction group increased(P<0.05).The expression ofα-SMA protein in the Supplemented Buyang Huanwu Decoction group decreased(P<0.05).Conclusion The effect of Supplemented Buyang Huanwu Decoction in the prevention and treatment of postoperative abdominal adhesions is exact,which mechanism may be by reducing Epithelial mesenchymal transition of peritoneal mesothelial cells,reducing the deposition of extracellular matrix,thereby reducing peritoneal adhesion.