Department of Pharmacy, The Second Xiangya Hospital, Central South University, Changsha, 410011, PR China; Institute of Clinical Pharmacy, The Second Xiangya Hospital, Central South University, Changsha, 410011, PR China.
Department of Pharmacy, Yiyang Central Hospital, Yiyang, 41300, PR China.
Eur J Pharmacol. 2019 Sep 5;858:172480. doi: 10.1016/j.ejphar.2019.172480. Epub 2019 Jun 20.
Bone formation refers to a series of complex events related to the activities of osteoblasts. In this study, we evaluated the osteogenesis activity of a natural compound named isocoumarin A that was isolated from the rhizomes of Polygonum amplexicaule on the non-transformed preosteoblastic cell line MC3T3-E1 for an in vitro study, and the results revealed that it increased the proliferation and promoted the mineralization of the extracellular matrix of MC3T3-E1 cells after treatment for 3 d in a dose-dependent manner. The cell metabolic activity peaked at 169% at 10 μM, and the activity of alkaline phosphatase (ALP) tripled to 15.94 U/mg compared with the control group. The protein levels of morphogenetic protein 2 (BMP-2), runt-related transcription factor 2 (RUNX2), ALP, and the mRNA levels of ALP, type I collagen (COL-1), and osteocalcin (OCN) were also upregulated after isocoumarin A administration. The mechanism investigation revealed that these effects were associated with the activation of the p-Akt/p-Erk1/2-activated BMP/RUNX2 signaling pathway. Subsequently, the in vivo investigation on the zebrafish embryos model demonstrated that isocoumarin A (0.30 mM) increased the number of vertebrae (5.38 ± 2.07 pcs) and the vertebral area (433.25 ± 111.77 μm) in the development process of zebrafish embryos after a 7-day postfertilization (dpf) culture compared with the control group (2.50 ± 1.16 pcs and 209.75 ± 86.40 μm). Together, these results indicated that isocoumarin A could be viewed as a promising candidate in early drug discovery and development to promote the healing of fractures and postmenopausal osteoporosis.
骨形成是指与成骨细胞活性相关的一系列复杂事件。在这项研究中，我们评估了从虎杖的根茎中分离得到的天然化合物异香豆素 A 对非转化前成骨细胞系 MC3T3-E1 的成骨活性，结果表明，它在体外能剂量依赖性地增加细胞增殖，并促进 MC3T3-E1 细胞外基质的矿化。在 10 μM 时，细胞代谢活性达到峰值，为对照组的 169%；碱性磷酸酶 (ALP) 的活性增加了两倍，达到 15.94 U/mg。与对照组相比，骨形态发生蛋白 2 (BMP-2)、 runt 相关转录因子 2 (RUNX2)、ALP 的蛋白水平以及 ALP、I 型胶原 (COL-1) 和骨钙素 (OCN) 的 mRNA 水平也上调。机制研究表明，这些作用与 p-Akt/p-Erk1/2 激活的 BMP/RUNX2 信号通路的激活有关。随后，在斑马鱼胚胎模型中的体内研究表明，与对照组相比，异香豆素 A (0.30 mM) 在受精后 7 天的胚胎发育过程中增加了斑马鱼胚胎的椎骨数量 (5.38 ± 2.07 pcs) 和椎骨面积 (433.25 ± 111.77 μm)。总之，这些结果表明，异香豆素 A 可以作为一种有前途的候选药物，用于早期药物发现和开发，以促进骨折和绝经后骨质疏松症的愈合。
