Cuijpers Iris, Sthijns Mireille M J P E, van den Bogart Veerle A R, Katsburg Joey, Leenders Cliff F M, Troost Freddy J
Department of Human Biology, Institute of Nutrition and Translational Research in Metabolism (NUTRIM), Maastricht University, 6200 MD Maastricht, The Netherlands.
Food Innovation and Health, Centre for Healthy Eating and Food Innovation (HEFI), Maastricht University Campus Venlo, 5911 BV Venlo, The Netherlands.
Int J Mol Sci. 2025 May 27;26(11):5151. doi: 10.3390/ijms26115151.
In fibrotic skeletal muscles, excessive extracellular matrix (ECM) deposition is a result of increased activation and decreased apoptosis of myofibroblasts. The aim of this study is to investigate whether treatment with quercetin, kaempferol or capsaicin can reduce the transforming growth factor-beta 1 (TGF-β1)-induced myofibroblast differentiation and fibrotic ECM expression in differentiated C2C12 cells. Two-day-differentiated C2C12 cells were treated with TGF-β1 for 48 h to induce myofibroblast differentiation. Twenty-four hours before (pre-treatment) and for forty-eight hours with (co-treatment) TGF-β1 treatment, cells were exposed to quercetin (25, 50 µM), kaempferol (10, 25, 50 µM) or capsaicin (25, 50 µM). The immunofluorescence intensity of alpha smooth muscle actin (αSMA) and collagen type I/III gene expression were assessed as myofibroblast markers. MyoD immunofluorescence intensity was measured as a myogenic marker. Co-treatment of TGF-β1 with the phytochemicals was most effective, resulting in a decreased number of αSMA-positive cells (all three compounds), decreased collagen type I (kaempferol, capsaicin) and type III (kaempferol) gene expression, and increased MyoD (kaempferol, capsaicin) protein expression compared to TGF-β1 treatment. This study demonstrates that treatment with quercetin, kaempferol or capsaicin can reduce myofibroblast markers. This suggests a possible anti-fibrotic effect of the phytochemicals in skeletal muscle.
在纤维化骨骼肌中，细胞外基质（ECM）过度沉积是成肌纤维细胞活化增加和凋亡减少的结果。本研究的目的是调查用槲皮素、山奈酚或辣椒素处理是否可以减少转化生长因子-β1（TGF-β1）诱导的分化C2C12细胞中的成肌纤维细胞分化和纤维化ECM表达。将分化两天的C2C12细胞用TGF-β1处理48小时以诱导成肌纤维细胞分化。在TGF-β1处理前24小时（预处理）以及处理的48小时内（共处理），将细胞暴露于槲皮素（25、50μM）、山奈酚（10、25、50μM）或辣椒素（25、50μM）。评估α平滑肌肌动蛋白（αSMA）的免疫荧光强度和I/III型胶原基因表达作为成肌纤维细胞标志物。测量MyoD免疫荧光强度作为肌源性标志物。TGF-β1与这些植物化学物质共同处理最为有效，与TGF-β1处理相比，导致αSMA阳性细胞数量减少（所有三种化合物）、I型胶原（山奈酚、辣椒素）和III型胶原（山奈酚）基因表达降低以及MyoD（山奈酚、辣椒素）蛋白表达增加。本研究表明，用槲皮素、山奈酚或辣椒素处理可以减少成肌纤维细胞标志物。这表明这些植物化学物质在骨骼肌中可能具有抗纤维化作用。
