背景:软骨细胞是软骨中唯一的细胞成分,再生能力差。因此,修复受损的软骨仍然是一个巨大的挑战。梅花鹿鹿角主要由软骨组织组成,具有惊人的修复和更新能力。我们先前的研究已经证明转化生长因子β(TGF-β1)被认为是参与快速生长的关键分子,在软骨层中表现最强。然而,鹿TGF-β1是否具有与小鼠等其他物种明显不同的功能,以及调节软骨生长的分子机制。
方法:从新生小鼠肋软骨中收集原代软骨细胞。通过RNA测序(RNA-seq)技术结合定量实时聚合酶链反应(qRT-PCR)和免疫荧光测定(IFA)等验证方法,阐明了TGF-β1对原代软骨细胞活力的影响。使用DEGseq包鉴定差异表达基因。
结果:我们的结果表明,鹿TGF-β1的过表达可能促进软骨细胞增殖和细胞外基质(ECM)的合成,同时通过调节转录因子抑制软骨细胞分化,生长因子,ECM相关基因,增殖和分化标记基因,如Comp,Fgfr3、Atf4、Stat1等。,和信号通路,如MAPK信号通路,TRP通道等的炎症介质调节。此外,通过比较鹿TGF-β1和小鼠TGF-β1的氨基酸序列和结构,我们发现鹿TGF-β1和小鼠TGF-β1蛋白在臂结构域上主要在结构上不同,这是主要的功能域。表型鉴定结果表明,鹿TGF-β1可能比小鼠TGF-β1具有更强的功能。
结论:这些结果表明,鹿TGF-β1具有通过调节软骨细胞增殖来促进软骨形成的能力,分化和ECM合成。这项研究提供了有关鹿TGF-β1对软骨细胞活力影响的分子机制的见解。
BACKGROUND: Chondrocytes are the only cell components in the cartilage, which has the poor regeneration ability. Thus, repairing damaged cartilage remains a huge challenge. Sika deer antlers are mainly composed of cartilaginous tissues that have an astonishing capacity for repair and renewal. Our previous study has demonstrated the transforming growth factor β (TGF-β1) is considered to be a key molecule involved in rapid growth, with the strongest expression in the cartilage layer. However, it remains to be clarified whether deer TGF-β1 has significantly different function from other species such as mouse, and what is the molecular mechanism of regulating cartilage growth.
METHODS: Primary chondrocytes was collected from new born mouse rib cartilage. The effect of TGF-β1 on primary chondrocytes viability was elucidated by RNA sequencing (RNA-seq) technology combined with validation methods such as quantitative real-time polymerase chain reaction (qRT-PCR) and immunofluorescence assay (IFA). Differential expression genes were identified using the DEGseq package.
RESULTS: Our results demonstrated that the overexpression of deer TGF-β1 possibly promoted chondrocyte proliferation and extracellular matrix (ECM) synthesis, while simultaneously suppressing chondrocyte differentiation through regulating transcription factors, growth factors, ECM related genes, proliferation and differentiation marker genes, such as Comp, Fgfr3, Atf4, Stat1 etc., and signaling pathways such as the MAPK signaling pathway, inflammatory mediator regulation of TRP channels etc. In addition, by comparing the amino acid sequence and structures between the deer TGF-β1 and mouse TGF-β1, we found that deer TGF-β1 and mouse TGF-β1 proteins are mainly structurally different in arm domains, which is the main functional domain. Phenotypic identification results showed that deer TGF-β1 may has stronger function than mouse TGF-β1.
CONCLUSIONS: These results suggested that deer TGF-β1 has the ability to promote chondrogenesis by regulating chondrocyte proliferation, differentiation and ECM synthesis. This study provides insights into the molecular mechanisms underlying the effects of deer TGF-β1 on chondrocyte viability.