Sonichedgehog(Shh)是Hedgehog信号通路的一个组成部分,在调节细胞增殖中起重要作用,分化,凋亡,以及受损生物的修复。为进一步阐明Shh基因在绒山羊次级毛囊生长周期中的表达规律及其对次级毛囊乳头细胞的作用机制,提高羊绒质量,在这项研究中,以内蒙古阿尔巴斯白绒山羊为研究对象,采集不同生长阶段的皮肤样本,通过RT-qPCR检测Shh及其基因表达,蛋白质印迹,免疫组织化学,和其他技术,同时我们还在体外培养了DPCs。构建Shh基因过表达和干扰载体,并通过细胞转染技术研究Shh基因对DPCs增殖和凋亡的影响。结果表明,绒山羊次级毛囊生长周期皮肤中Shh及其基因表达存在显著差异,在生长期中表达水平最高,紧随其后的是卡特金,和调音素的最低表达水平。Shh主要表现在根鞘内,外根鞘,和继发性毛囊乳头。Shh基因过表达后,与干扰组相比,毛乳头细胞的增殖和活力增强。Shh基因干扰后,细胞的凋亡率增加,表明Shh基因可以调节下游Ptch,Smo,和Gli2基因表达促进DPCs的增殖,从而形成其在绒山羊次级毛囊生长周期中的表达模式。
Sonic hedgehog (
Shh) is a component of the Hedgehog signaling pathway, playing an important role in regulating cell proliferation, differentiation, apoptosis, and the repair of damaged organisms. To further clarify the expression pattern of
Shh gene in the secondary hair follicle growth cycle of cashmere goats and its mechanism of action on secondary hair follicle papilla cells, and improve cashmere quality, in this study, we took Inner Mongolia Albas white cashmere goats as the research objects and collected skin samples at different growth stages to obtain secondary hair follicles, detected
Shh and its gene expression by RT-qPCR, Western blot, immunohistochemistry, and other techniques, while we also cultured DPCs in vitro.
Shh gene overexpression and interference vectors were constructed, and the effects of
Shh gene on the proliferation and apoptosis of DPCs were studied through cell transfection technology. The results showed that there are significant differences in Shh and its gene expression in the secondary hair follicle growth cycle skins of cashmere goats, with the highest expression level in anagen, followed by catagen, and the lowest expression level in telogen. Shh was mainly expressed in the inner root sheath, outer root sheath, and secondary hair follicle papilla. After the overexpression of Shh gene, the proliferation and vitality of the hair papilla cells were enhanced compared to the interference group. After
Shh gene interference, the apoptosis rate of the cells increased, indicating that
Shh gene can regulate downstream Ptch, Smo, and Gli2 gene expression to promote the proliferation of DPCs, and thus form its expression pattern in the secondary hair follicle growth cycle of cashmere goats.