Jin Mei, Qiu Xinyue, Piao Jing'ai, Zhang Lijuan, Piao Jun, Zhao Fengqin
Liaoning Provincial Key Laboratory of Biotechnology and Drug Discovery, Department of Life Sciences, Liaoning Normal University, Dalian, China.
Anim Biotechnol. 2022 Nov;33(6):1255-1267. doi: 10.1080/10495398.2021.1886940. Epub 2021 Mar 28.
In this study, the genes related to the Downy growth of Liaoning cashmere goats were screened for their expression with simultaneous melatonin administration, so as to investigate the effects of target genes on the proliferation of skin fibroblasts in this animal species. Genes related to the villus growth of skin fibroblasts were screened by in vitro transcriptome sequencing and verified by qPCR. In addition, gene overexpression and interference were used to study the effects of target genes on the proliferation of skin fibroblasts. Groups treated with M1_24H, M2_24H and M2_72H exhibited significant differences compared with the control group. Among them, the differentially expressed transcripts in the M2_72H group were significantly enriched in the TNF and NOD-like receptor signaling pathways, which are associated with the villus. In addition, eight differentially expressed genes were screened from the TNF and the NOD-like receptor signaling pathways. Verification by qPCR showed that the expression of TNF-α, IL-6, TNFAIP3, PYCARD and NFKBIA genes were significantly upregulated, which was consistent with the sequencing results. Melatonin treatments can significantly lead to an increase in the expression of IL-6 and TNF-α genes. Besides, melatonin treatments can affect cashmere growth in Liaoning cashmere goats by regulating several signaling pathways, including TNF, NOD-like receptor and NF-κB.
在本研究中,筛选了与辽宁绒山羊绒毛生长相关的基因,以研究褪黑素同时给药时它们的表达情况,从而探究目标基因对该物种皮肤成纤维细胞增殖的影响。通过体外转录组测序筛选与皮肤成纤维细胞绒毛生长相关的基因,并通过qPCR进行验证。此外,利用基因过表达和干扰来研究目标基因对皮肤成纤维细胞增殖的影响。与对照组相比,M1_24H、M2_24H和M2_72H处理组表现出显著差异。其中,M2_72H组中差异表达的转录本在与绒毛相关的TNF和NOD样受体信号通路中显著富集。此外,从TNF和NOD样受体信号通路中筛选出8个差异表达基因。qPCR验证表明,TNF-α、IL-6、TNFAIP3、PYCARD和NFKBIA基因的表达显著上调,这与测序结果一致。褪黑素处理可显著导致IL-6和TNF-α基因表达增加。此外,褪黑素处理可通过调节包括TNF、NOD样受体和NF-κB在内的多种信号通路来影响辽宁绒山羊的羊绒生长。