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目的探讨龟板提取物(Plastrum Testudinis extracts,PTE)对大鼠触须部皮肤创面修复中Wnt/β-catenin通路的调控。方法分离培养SD乳鼠毛囊干细胞(Hair follicle stem cells,HFSCs),免疫荧光检测细胞表面标志物CD34、CD71表达,CCK-8法检测PTE对HFSCs增殖的影响,Western-blot检测PTE对HFSCs中Gsk-3β、β-catenin蛋白表达的作用;制作SD大鼠触须部皮肤缺损模型,随机分为空白对照组、实验对照组、阳性药物对照组、PTE组,测算动物创面面积和愈合率,病理学及免疫荧光检测创面组织。结果培养的细胞形态饱满,折光性强,呈铺路石样,细胞表面CD34及CD71阳性。CCK-8显示PTE对HFSCs有促增殖作用(P<0.01)。PTE作用于HFSCs后,Gsk-3β蛋白(P<0.05)下降的同时β-catenin蛋白表达上升(P<0.001)。术后7 d各组创面基本愈合,PTE组及阳性药物对照组表皮较其他组薄,毛囊周围有新生表皮参与修复,表皮与真皮结合牢固,真皮无炎性细胞,组织结构排列规则,促修复效果优于其他各组;PTE组修复表皮内Gsk-3β蛋白下降而β-catenin蛋白表达上升。结论 PTE可通过激活HFSCs中Wnt/β-catenin通路促进SD大鼠触须部创面修复。
Objective To investigate the regulation of Wnt / β-catenin pathway in repair of the skin wounds in the antenna of rat sensilla by Plastrum Testudinis extracts (PTE). Methods Hair follicle stem cells (HFSCs) were isolated and cultured. The expression of cell surface markers CD34 and CD71 were detected by immunofluorescence. The effect of PTE on the proliferation of HFSCs was detected by CCK-8. The effect of PTE on the expression of Gsk 3β, β-catenin protein expression in the skin of SD rats were made. The rats were randomly divided into blank control group, experimental control group, positive drug control group and PTE group. The wound area and healing rate were calculated. Pathology And immunofluorescence detection of wound tissue. Results The cultured cells were full of shape, strong refraction and were paved with paving stones, and the cell surface was positive for CD34 and CD71. CCK-8 showed that PTE promoted proliferation of HFSCs (P <0.01). After PTE treatment, the expression of β-catenin increased (P <0.001) while Gsk-3β (P <0.05) decreased. The wounds of all the groups basically healed on the 7th day. The epidermis of the PTE group and the positive drug control group was thinner than the other groups. The epidermis around the hair follicles was involved in the repair. The epidermis and the dermis were firmly bonded. The dermis had no inflammatory cells. The effect was better than other groups. PTE group decreased Gsk-3β protein and β-catenin protein expression in the epidermis. Conclusion PTE can promote Wnt / β-catenin pathway in HFSCs and promote the repair of the wounds in the constrictors of the SD rats.