傣药咪多领(云南琵琶甲)对增生性瘢痕中表皮干细胞干性及Hedgehog信号通路的影响

目的探讨傣药咪多领(云南琵琶甲)对增生性瘢痕中表皮干细胞干性及Hedgehog信号通路的影响。方法构建新西兰大耳兔增生性瘢痕模型,并分离增生性瘢痕组织中表皮干细胞。将动物随机分为对照组、模型组、阳性对照组(PC组,用复方肝素钠尿囊素凝胶干预)及傣药咪多领0.667、1.334、2.668 g/kg组;表皮干细胞分为对照组,傣药咪多领0.025、0.05、0.1 g/mL组。RT-qPCR和Western blot测各组表皮干细胞标志物(Nanog和Oct4)及Hedgehog信号通路相关基因(SHH、Patch、Smo和Gli-1)mRNA和蛋白的表达水平,进一步通过免疫荧光检测SHH、Patch、Smo和Gli-1的表达水平。CCK-8法检测表皮干细胞增殖活力。流式细胞仪检测各组CD49F、CD29和CK19阳性细胞比率。结果傣药咪多领对增生性瘢痕具有良好的治疗效果,且显著抑制增生性瘢痕组织中Nanog和Oct4及SHH、Patch、Smo和Gli-1的表达水平,此外,2.668 g/kg剂量傣药咪多领与15 mg/kg剂量复方肝素钠尿囊素凝胶治疗效果相近。傣药咪多领限制抑制表皮干细胞中Nanog和Oct4及SHH、Patch、Smo和Gli-1的表达水平。结论增生性瘢痕中表皮干细胞干性增强,且Hedgehog信号通路相关蛋白表达上调,而傣药咪多领可逆转这一过程,并对增生性瘢痕具有良好的治疗效果。

The Influence Mechanism of the Blaps Rynchopetera Fairmaire on the Stemness of Epidermal Stem Cells and Hedgehog Signaling Pathway in Hypertrophic Scar

Objective To explore the effect of Blaps rynchopetera Fairmaire on the stemness of epidermal stem cells and Hedgehog signaling pathway in hypertrophic scar.Methods Model of hypertrophic scar was constructed,and the epidermal stem cells in the hypertrophic scar tissue were sorted.Animal were divided into control group,model group,positive control group(PC,intervention with compound heparin sodium and allantoin gel)and Blaps rynchopetera Fairmaire 0.667 g/kg,1.334 g/kg,2.668 g/kg groups.Epidermal stem cells were divided into control group and Blaps ryhchopetera Fairmaire 0.025 g/mL,0.05 g/mL,0.1 g/mL groups.Respectively,RT-qPCR and Western blot were used to detect the expression levels of mRNA and protein of epidermal stem cell markers(Nanog and Oct4)and Hedgehog signaling pathway related genes(SHH,Patch,Smo and Gli-1)in each group.Further,the expression levels of SHH,Patch,Smo and Gli-1 were detected by immunofluorescence.Flow cytometry detected the rate of CD49F,CD29 and CK19 positive cells in each group.Results The Blaps rynchopetera Fairmaire had a good therapeutic effect on hypertrophic scar,and significantly inhibited the expression levels of Nanog,Oct4,SHH,Patch,Smo and Gli-1 in hypertrophic scar tissues.In addition,the therapeutic effect of the Blaps rynchopetera Fairmaire at dose of 2.668 g/kg was similar to that of compound heparin sodium and allantoin gel at a dose of 15 mg/kg.Blaps rynchopetera Fairmaire inhibited the expression levels of Nanog and Oct4,SHH,Patch,Smo and Gli-1 in epidermal stem cells.Conclusion In hypertrophic scars,the stemness of epidermal stem cells was enhanced,and the expression of Hedgehog pathway-related proteins was upregulated.The Blaps rynchopetera Fairmaire reverses this process and had good therapeutic effect on hypertrophic scars.