Division of Exercise Science and Sports Medicine, Department of Human Biology, University of Cape Town, South Africa; UCT Research Centre for Health through Physical Activity, Lifestyle and Sport, South Africa; International Federation of Sports Medicine (FIMS) Collaborative Centre of Sports Medicine, South Africa.
Division of Cell BiologyDepartment of Human Biology, University of Cape Town, South Africa.
J Sci Med Sport. 2020 Aug;23(8):695-700. doi: 10.1016/j.jsams.2020.02.007. Epub 2020 Feb 8.
To investigate the functional effect of implicated variants within BGN and COL5A1 on gene expression of components of the extracellular matrix (ECM) in a TGF-β-stimulated risk model for musculoskeletal soft tissue injuries.
Experimental research, laboratory study.
Skin biopsies were obtained from nine healthy participants with either a combined increased or reduced risk profile for COL5A1 rs12722 C>T and BGN rs1126499 C>T - rs1042103 G>A, and primary fibroblast cell lines were established. Total RNA was extracted at baseline (10% FBS), after serum starvation (1% FBS) and TGF-β1 treatment (1% FBS, 10ng/mL TGF-1β). Relative mRNA levels of BGN, COL5A1, DCN and VEGFA was quantified using Taqman® array pre-spotted plate assays (Applied Biosystems, Foster city, CA, USA).
At baseline, the reduced risk group had 2.5, 1.9 and 2 fold increases (p<0.001) in relative BGN, COL5A1 and VEGFA mRNA levels respectively. In the serum starved experiments, except for a significant 1.5 fold (p=0.017) increase in relative DCN mRNA expression in the reduced risk group, similar observations were noted for the other three genes. After TGF-1β treatment, the reduced risk group had 1.3 (p=0.011) and 1.4 fold (p=0.001) increases in the relative COL5A1 and VEGFA mRNA levels, respectively.
Altered mRNA levels associated with genetic risk profiles for musculoskeletal soft injury risk at baseline (BGN, COL5A1 and VEGFA), with serum starvation (DCN) and after TGF-β1 treatment (COL5A1 and VEGFA) provide additional functional evidence to support the association of implicated genetic loci with several musculoskeletal soft tissue injuries. Implication of altered gene expression profiles underpinning these genetic risk associated loci potentially highlight key therapeutic targets for management of these injuries.
研究 TGF-β刺激的肌肉骨骼软组织损伤风险模型中 BGN 和 COL5A1 内受影响的变异对细胞外基质 (ECM) 成分基因表达的功能影响。
实验研究，实验室研究。
从九名健康参与者中获取皮肤活检样本，这些参与者具有 COL5A1 rs12722 C>T 和 BGN rs1126499 C>T-rs1042103 G>A 联合增高或降低风险特征，并建立原代成纤维细胞系。在基线（10% FBS）、血清饥饿（1% FBS）和 TGF-β1 处理（1% FBS，10ng/mL TGF-β1）后提取总 RNA。使用 Taqman®array 预点样板检测试剂盒（Applied Biosystems，加利福尼亚州福斯特市）定量测定 BGN、COL5A1、DCN 和 VEGFA 的相对 mRNA 水平。
在基线时，低风险组的 BGN、COL5A1 和 VEGFA mRNA 水平分别增加了 2.5、1.9 和 2 倍（p<0.001）。在血清饥饿实验中，除了低风险组的 DCN mRNA 表达有显著的 1.5 倍（p=0.017）增加外，其他三个基因也观察到了类似的结果。TGF-β1 处理后，低风险组的 COL5A1 和 VEGFA mRNA 水平分别增加了 1.3（p=0.011）和 1.4 倍（p=0.001）。
与肌肉骨骼软组织损伤风险的遗传风险谱相关的基因表达水平的改变（BGN、COL5A1 和 VEGFA），在血清饥饿（DCN）和 TGF-β1 处理后（COL5A1 和 VEGFA）提供了额外的功能证据，支持了受影响遗传位点与几种肌肉骨骼软组织损伤的相关性。这些遗传风险相关位点下改变的基因表达谱的意义，可能突出了这些损伤管理的关键治疗靶点。
