Academician Workstation, Jiangxi University of Chinese Medicine, Nanchang, 330004, P. R. China.
Department of Medicine, Zhejiang Academy of Traditional Chinese Medicine, Hangzhou, 310007, P. R., China.
Chem Biodivers. 2024 Feb;21(2):e202301383. doi: 10.1002/cbdv.202301383. Epub 2024 Jan 25.
Herba Epimedii is widely used to promote bone healing, and their active ingredients are total flavonoids of Epimedium (TFE). Ras homolog gene family member A / Rho-associated protein kinase (RhoA/Rock), an important pathway regulating the cytoskeleton, has been proven to affect bone formation. However, whether TFE promotes bone healing via this pathway remains unclear. In this study, the therapeutic effects of TFE were estimated using micro-computed tomography and hematoxylin and eosin staining of pathological sections. F-actin in osteoblasts was stained to investigate the protective effects of TFE on the cytoskeleton. Its regulatory effects on the RhoA/Rock1 pathway were explored using RT-qPCR and Western blot analysis. Besides, flow cytometry, alkaline phosphatase and nodule calcification staining were performed to evaluate the effects on osteogenesis. The bone healing in rats was improved, the cytoskeletal damage in osteoblasts was reduced, the RhoA/Rock1 pathway was downregulated, and osteogenesis was enhanced after TFE treatment. Thus, TFE can promote bone formation at least partially by regulating the expression of key genes and proteins in the cytoskeleton. The findings of this study provided evidence for clinical applications and would contribute to a better understanding of Epimedium's mechanisms in treating bone defects.
淫羊藿被广泛用于促进骨愈合,其有效成分是淫羊藿总黄酮(TFE)。Ras 同源基因家族成员 A/Rho 相关蛋白激酶(RhoA/Rock),一种调节细胞骨架的重要途径,已被证明影响骨形成。然而,TFE 是否通过该途径促进骨愈合尚不清楚。在这项研究中,通过 micro-CT 和苏木精-伊红染色的病理切片来评估 TFE 的治疗效果。用 F-肌动蛋白染色来研究 TFE 对细胞骨架的保护作用。通过 RT-qPCR 和 Western blot 分析探讨其对 RhoA/Rock1 通路的调节作用。此外,通过流式细胞术、碱性磷酸酶和结节钙化染色来评估其对成骨的影响。TFE 治疗可改善大鼠的骨愈合,减轻成骨细胞的细胞骨架损伤,下调 RhoA/Rock1 通路,增强成骨作用。因此,TFE 可以通过调节细胞骨架关键基因和蛋白的表达至少部分地促进骨形成。本研究结果为临床应用提供了证据,并有助于更好地理解淫羊藿在治疗骨缺损中的作用机制。