摘要
旨在探索褪黑激素(melatonin, MT)对let-7家族成员及其靶基因在内蒙古绒山羊皮肤周期性表达的影响。本研究选取内蒙古绒山羊罕山型个体6只,分为埋植组(皮下埋植MT,n=3)和对照组(不埋植MT,n=3),连续12个月采集绒山羊个体皮肤组织为试验材料,利用qRT-PCR技术检测皮肤组织中11个let-7家族成员的表达变化,结合RNA-Seq技术分析let-7家族靶基因的表达变化。结果表明:1)在整个皮肤毛囊周期内,MT上调let-7b、let-7e、let-7a-3p的表达,下调let-7f、let-7g、let-7c、let-7a-5p、let-7d、miR-98的表达。2)在4月份,MT上调"let-7g、let-7i、miR-98"的表达,下调PTGS2、PLCG1等靶基因的表达;下调"let-7a-5p、let-7b、let-7c"的表达,上调TGIF2、RBL1等靶基因的表达。3)在6月份,MT下调"let-7g、let-7i、miR-98"的表达,上调PTGS2、PLCG1、SRC等靶基因的表达;上调"let-7a-5p、let-7b、let-7c"的表达,下调GDF5、TGIF2、RBL1等靶基因的表达。4)let-7家族成员的靶基因主要参与VEGF、TGF-β、Oxytocin和NF-kappa B信号通路,在细胞过程、激素调节和转录因子调控方面发挥重要作用。综上表明,MT可能通过改变let-7家族重要成员的表达水平,影响相应靶基因的表达,进而在皮肤毛囊的周期性生长过程中发挥重要作用,研究结果为进一步揭示MT介导miRNAs影响羊绒生长的分子机理提供了理论依据。
The study aimed to explore the effects of melatonin on the periodic expression of let-7 family and its target genes in skin of Inner Mongolia Cashmere goat. In this study, six Inner Mongolia Hanshan Cashmere goats were selected and divided equally and randomly into 2 groups, melatonin implantation group and non-implantation group. Taking the skin tissue of the goats as experimental material in every months of the year. The expression changes of 11 members in let-7 family were quantified by reverse transcription-real time polymerase chain reaction(qRT-PCR) in the skin tissue. The expression changes of the target genes of let-7 family were analyzed using the RNA-Seq technique. The results showed that: 1) During the whole hair follicle cycle, melatonin up-regulated the expression of let-7 b, let-7 e and let-7 a-3 p, down-regulated the expression of let-7 f, let-7 g, let-7 c, let-7 a-5 p, let-7 d and miR-98. 2) In April, melatonin up-regulated the expression of "let-7 g, let-7 i and miR-98", down-regulated the expression of target genes such as PTGS2 and PLCG1. It down-regulated the expression of "let-7 a-5 p, let-7 b and let-7 c", up-regulated the expression of target genes such as TGIF2 and RBL1. 3) In June, melatonin down-regulated the expression of "let-7 g, let-7 i and miR-98", up-regulated the expression of target genes such as PTGS2, PLCG1 and SRC; It up-regulated the expression of "let-7 a-5 p, let-7 b and let-7 c", down-regulated the expression of target genes such as GDF5, TGIF2 and RBL1. 4) The target genes of let-7 family were mainly involved in VEGF, TGF-β, Oxytocin and NF-kappa B signaling pathways, and played an important role in cell processes, hormone regulation and regulation of transcription factors. In summary, MT may affect the expression of the corresponding target genes by changing the expression level of members in the let-7 family. It plays an important role in the periodic growth of skin hair follicles. These results provided a theoretical basis for further revealing the molecular mechanism of melatonin mediating miRNAs to affect cashmere growth.
引文
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