Romagnoli Cecilia, Zonefrati Roberto, Lucattelli Elena, Innocenti Marco, Civinini Roberto, Iantomasi Teresa, Brandi Maria Luisa
Department of Experimental and Clinical Biomedical Sciences, University of Florence, 50139 Florence, Italy.
Italian Foundation for the Research on Bone Diseases (F.I.R.M.O.), 50129 Florence, Italy.
Biomedicines. 2023 Mar 27;11(4):1017. doi: 10.3390/biomedicines11041017.
Parathyroid hormone (PTH) is a hormone secreted by the parathyroid glands. Despite its well-known characterized anabolic and catabolic actions on the skeleton, the in vitro effects of PTH on skeletal muscle cells are limited and generally performed on animal models. The aim of this study was to evaluate the effects of a short impulse of PTH (1-84) on the proliferation and the differentiation of skeletal muscle satellite cells isolated from human biopsies. The cells were exposed for 30 min to different concentrations of PTH (1-84), from 10 mol/L to 10 mol/L. ELISA was used to assay cAMP and the myosin heavy-chain (MHC) protein. The proliferation was assayed by BrdU and the differentiation by RealTime-qPCR. A statistical analysis was performed by ANOVA followed by Bonferroni's test. No significant variations in cAMP and the proliferation were detected in the isolated cells treated with PTH. On the other hand, 10 mol/L PTH on differentiated myotubes has shown significant increases in cAMP ( ≤ 0.05), in the expression of myogenic differentiation genes ( ≤ 0.001), and in the MHC protein ( ≤ 0.01) vs. untreated controls. This work demonstrates for the first time the in vitro effects of PTH (1-84) on human skeletal muscle cells and it opens new fields of investigation in muscle pathophysiology.
甲状旁腺激素(PTH)是由甲状旁腺分泌的一种激素。尽管其对骨骼的合成代谢和分解代谢作用已广为人知,但PTH对骨骼肌细胞的体外作用有限,且通常在动物模型上进行研究。本研究的目的是评估甲状旁腺激素(1-84)短脉冲对从人体活检组织中分离出的骨骼肌卫星细胞增殖和分化的影响。将细胞暴露于浓度为10⁻¹¹mol/L至10⁻⁷mol/L的不同浓度甲状旁腺激素(1-84)中30分钟。采用酶联免疫吸附测定法(ELISA)检测环磷酸腺苷(cAMP)和肌球蛋白重链(MHC)蛋白。通过5-溴脱氧尿嘧啶核苷(BrdU)检测增殖情况,通过实时定量聚合酶链反应(RealTime-qPCR)检测分化情况。采用方差分析(ANOVA),随后进行邦费罗尼检验进行统计分析。在用甲状旁腺激素处理的分离细胞中,未检测到cAMP和增殖的显著变化。另一方面,与未处理对照相比,10⁻⁷mol/L甲状旁腺激素对分化的肌管显示出cAMP显著增加(P≤0.05)、生肌分化基因表达显著增加(P≤0.001)以及MHC蛋白显著增加(P≤0.01)。这项工作首次证明了甲状旁腺激素(1-84)对人骨骼肌细胞的体外作用,并为肌肉病理生理学开辟了新的研究领域。