探讨皮粘散调控Wnt4/β-catenin信号通路促进慢性皮肤溃疡小鼠创面愈合的作用机制  

Mechanism of Pizhan Powder in regulating the Wnt4/β-catenin signaling pathway to promote wound healing in mice with chronic skin ulcers

作　　者：阙平鑫毅 肖湘 曾立 赵显斌 肖敏[3] 唐宋琪 QUE Pingxinyi;XIAO Xiang;ZENG Li;ZHAO Xianbin;XIAO Min;TANG Songqi(School of Pharmacy,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China;Hainan Medical University,Haikou5.71199,China;Chengdu University of Traditional Chinese Medicine,Chengdu 610075,China)

机构地区：[1]成都中医药大学药学院,成都611137 [2]海南医科大学 [3]成都中医药大学

出　　处：《北京中医药大学学报》2025年第2期205-215,共11页Journal of Beijing University of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(No.82004380)。

摘　　要：目的 探讨皮粘散调控Wnt4/β-连环蛋白(β-catenin)信号通路促进慢性皮肤溃疡小鼠创面愈合的作用机制。方法 雄性BALB/c小鼠按随机数字表法分为空白组、模型组、皮粘散组、皮粘散去皮组、皮粘散皮部组、抑制剂组、皮粘散+抑制剂组,每组6只。除空白组外,其余各组采用埋置异物法制备慢性皮肤溃疡模型。造模术后,模型组以呋喃西林溶液清理创面,皮粘散组、皮粘散去皮组、皮粘散皮部组分别在小鼠皮肤创面涂抹对应药物0.1 g;抑制剂组腹腔注射3-(4-甲基苯基磺酰胺基)苯甲酸甲酯(MSAB)10 mg/kg;皮粘散+抑制剂组在小鼠皮肤创面涂抹皮粘散0.1 g,并腹腔注射MSAB(10 mg/kg)。每日1次,连续14 d。空白组不作处理。于给药的第1、3、7、14天记录创面愈合情况;HE染色法观察创面组织病理变化;免疫荧光法检测创面角蛋白10(CK10)、角蛋白14(CK14)、细胞增殖核抗原(Ki-67)、α-平滑肌肌动蛋白(α-SMA)和β-catenin表达;蛋白质印迹法检测Wnt4及β-catenin蛋白表达情况。结果 与模型组比较,皮粘散组创面面积缩小,创面愈合率升高(P<0.05);CK10、CK14、Ki-67、α-SMA、β-catenin及Wnt4蛋白表达增加(P<0.05)。与皮粘散组比较,皮粘散皮部组和皮粘散去皮组创面愈合率降低(P<0.05)。皮粘散去皮组创面愈合率及CK10、CK14、Ki-67、α-SMA蛋白荧光表达均低于皮粘散皮部组(P<0.05)。与皮粘散组比较,皮粘散+抑制剂组创面愈合率降低,CK10、CK14、Ki-67、α-SMA、β-catenin及Wnt4蛋白表达降低(P<0.05)。结论 皮粘散可通过调节Wnt4/β-catenin信号通路促进小鼠慢性皮肤溃疡创面愈合,其中皮部药物(牛皮、桑白皮、地骨皮)在促进创面愈合的过程中发挥了关键作用,是中医学“以皮治皮”理论的具体应用。Objective We aimed to explore the mechanism of Pizhan Powder in regulating the Wnt4/β-catenin signaling pathway to promote wound healing in mice with chronic skin ulcer.Methods Male BALB/c mice were divided into blank,model,Pizhan Powder,Pizhan powder removed bark medications,bark medications,inhibitor,and Pizhan Powder + inhibitor groups using the random number table method,with six mice per group.Except for the blank group,chronic skin ulcer wound models were prepared in the other groups by implanting foreign bodies.The blank control group received no treatment,whereas the wounds of the model group were cleaned with furacilin solution.The Pizhan Powder,Pizhan Powder removed bark medications,and bark medications groups were each administered 0.1 g of the corresponding medication on the skin wounds.The inhibitor group received an intraperitoneal injection of 3-(4-methylphenylsulfonamido) benzoic acid methyl ester(MSAB) at a dosage of 10 mg/kg.The Pizhan Powder + inhibitor group was administered 0.1 g of Pizhan Powder on the skin wound,and an intraperitoneal injection of MSAB was also administered(10 mg/kg).These treatments were administered once a day for 14 consecutive days.Wound healing was observed on the first,third,seventh,and 14th day of treatment;hematoxylin and eosin staining was used to observe the pathological changes of ulcerated skin;keratin 10(CK10),keratin 14(CK14),cell proliferation nuclear antigen(Ki-67),α-smooth muscle actin(α-SMA),and β-catenin expression in wounds was observed through immunofluorescence;Western blotting was used to detect the expression of signaling pathway-related proteins(Wnt4 and β-catenin).Results Compared to the model group,the Pizhan Powder group showed a reduced wound area and an increased wound healing rate(P<0.05) and elevated CK10,CK14,Ki-67,α-SMA,β-catenin,and Wnt4 protein expressions(P<0.05).Compared to the Pizhan Powder group,the wound healing rate of the bark medications and Pizhan Powder removed bark medications groups was reduced(P<0.05).The wound healing

关 键 词：慢性皮肤溃疡 皮粘散 创面愈合 Wnt4/β-连环蛋白信号通路 法象药理 小鼠 

分 类 号：R285.5[医药卫生—中药学]