2022, Vol. 26 ›› Issue (25): 3937-3943

Huo Xue Tong Luo capsule improves osteoblastogenesis of bone marrow mesenchymal stem cells through the ERα-Wnt/β-catenin signaling pathway

Wu Zhongshu1, Wei Yurou2, Chen Xiaojun1, Wuri Shana2, He Wei3, 4, Wei Qiushi3, 4   

Abstract: BACKGROUND: The Huo Xue Tong Luo capsule has been found to prevent the progression of asymptomatic osteonecrosis of the femoral head, but the mechanism is not clear.
OBJECTIVE: To investigate the effect of Huo Xue Tong Luo on the osteoblast differentiation of human bone marrow mesenchymal stem cells, and the effect of ERα-Wnt/β-catenin pathway.
METHODS: The third generation of human bone marrow mesenchymal stem cells was cultured in different concentrations of Huo Xue Tong Luo capsule (0, 1, 5, 10 mg/L) for 14 days. Alizarin red staining and alkaline phosphatase activity assay were used to determine the optimal concentration of Huo Xue Tong Luo in promoting osteogenic differentiation of human bone marrow mesenchymal stem cells. Human bone marrow mesenchymal stem cells were further divided into four groups: control group, Huo Xue Tong Luo group, Huo Xue Tong Luo + ICI 182780 group, and Huo Xue Tong Luo + ICI 182780 + DKK1 group. qRT-PCR, western blot assay, and immunofluorescence staining were used to investigate the effect of Huo Xue Tong Luo, ICI 182780 and recombinant human DKK1 protein on osteogenesis of human bone marrow mesenchymal stem cells.
RESULTS AND CONCLUSION: (1) Huo Xue Tong Luo capsule (10 mg/L) could significantly promote the proliferation and osteogenic differentiation of human bone marrow mesenchymal stem cells, and promote the mRNA expression of osteogenic genes Runx2, OPN, DLX5, BGLAP, and ERα, as well as the mRNA expression of Wnt/β-catenin pathway related targets β-catenin, TCF7, Lef1, C-MYC, CYCLIN and C-JUN. (2) The osteogenesis of Huo Xue Tong Luo capsule was inhibited by ICI 182780 and DKK1. (3) The results showed that ERα-Wnt/β-catenin signaling pathway was involved in the promoting effect of Huo Xue Tong Luo capsule on the proliferation and osteogenic differentiation of human bone marrow mesenchymal stem cells.
Key words: stem cells, bone marrow mesenchymal stem cells, hormone-induced femoral head necrosis, Huo Xue Tong Luo capsule, ERα, Wnt, β-catenin, pathway