褪黑素对绒山羊绒毛周期性生长相关基因CSDC2、FGF21、RORα表达的影响

Effect of Melatonin on Genes Including CSDC2,FGF21 and RORα Related to Hair Cycle Growth in Fibroblasts of Cashmere Goat Skin

绒山羊的绒毛生长具有周期性生长规律,埋置褪黑素可显著增加产绒量。以体外培养的绒山羊皮肤成纤维细胞为研究对象,探索褪黑素调控绒毛周期性生长的分子作用机制。采用组织块培养法培养绒山羊皮肤成纤维细胞,添加浓度为0 ng/L和500 ng/L的褪黑素处理72 h后经实时荧光定量PCR检测CSDC2、FGF21、RORα基因的表达量。结果表明,胎羊组织块自培养1 d后细胞沿边缘爬出,5 d后汇合度达80%;成年羊组织块于培养5~7 d后细胞爬出,14 d后细胞汇合度达70%,经鉴定为成纤维细胞,且胎羊成纤维细胞增殖能力更强,更适合后续功能实验的验证。经褪黑素处理后,CSDC2、FGF21表达水平显著高于对照组(P<0.05);RORα的表达量低于对照组但差异不显著(P>0.05)。说明褪黑素对绒毛周期性生长相关基因CSDC2、FGF21和RORα具有调控作用。

Cashmere growth in cashmere goats has a seasonal-cycle changes, and melatonin implant will increase the yield of cashmere. To explored how melatonin regulates the periodic growth of cashmere in vivo, this study identified the expression of key genes related to hair cycle growth for cashmere goat by melatonin induced in cashmere goat skin fibroblasts. The fibroblasts were cultured by tissue block culture method which was collected from cashmere goats, after cells identification, the fibroblasts were cultured with 0 ng/L and 500 ng/L concentration of melatonin, RT-qPCR were used to confirm the expression of CSDC2,FGF21 and RORα. The results showed that the fibroblasts of fetal goat skin grew from the tissue mass after 1 d, and the cells density reached 80% after 5 days;the adult goat fibroblasts grew after 5-7 days, and the confluence of cells reached 70% after 14days. Although both group of cells were identified as fibroblasts, the fetal fibroblasts demonstrate much better proliferational vitality and were more suitable for subsequent functional experiments. After treated melatonin, the mRNA expression level of CSDC2 and FGF21 was higher significantly than that in control group(P<0.05). Meanwhile, the expression level of RORα was lower than control group but had no significant difference(P>0.05). In conclusions, the melatonin can regulate the expression level of CSDC2,FGF21 and RORα.