Tang Yanghua, Mu Zhuosong, Pan Dong, Liu Renqi, Hong Shenghu, Xiong Zhenfei
Department of Orthopedics, Hangzhou Xiaoshan Hospital of Traditional Chinese Medicine, No. 156, Yucai Road, Xiaoshan District, Hangzhou, 311201, Zhejiang, China.
Third Clinical Medical College, Zhejiang Chinese Medical University, Binjiang District, Hangzhou, 310053, Zhejiang, China.
J Mol Histol. 2024 Oct;55(5):875-893. doi: 10.1007/s10735-024-10238-w. Epub 2024 Aug 6.
Osteoporosis is a metabolic bone disease. β-Catenin is associated with fractures. Jian-Pi-Bu-Shen (JPBS) can promote the healing of osteoporotic fractures (OPF). However, the mechanism of β-catenin-mediated skeletal muscle satellite cells (SMSCs) in OPF by the JPBS is unclear. SMSCs were isolated and divided into five groups. The results showed that the survival rate of SMSCs was significantly higher in the low, medium, and high dose JPBS-containing serum groups after 7 days of incubation. The ALP activity and the number of SMSCs mineralized in the JPBS-containing serum intervention group were elevated. Axin, GSK-3β, β-catenin siRNAs were constructed and transfected into cells. Transfection of siRNAs reduced Axin, GSK-3β, and β-catenin expressions, respectively. β-Catenin-siRNA reversed ALP activity, the number of SMSCs mineralized, and the expression of β-catenin, BMP2, Runx2, COL-I, SP7/Ostrix, Osteocalcin, and BMP-7. Transcriptomic results suggested that the TNF signaling pathway associated with OPF was enriched. SD rats were subjected to the construction of OPF model by removing the ovaries. JPBS decreased the levels of PINP, ALP, CTX, and NTX through β-catenin in OPF rats, while increasing Runx2, β-catenin expressions through β-catenin at the broken end of fractures. Moreover, JPBS decreased BMC, BMD, and BV/TV and improved pathological damage through β-catenin in OPF rats. JPBS decreased the expression of Axin, GSK-3β mRNA, and protein, but increased the expressions of β-catenin, Pax7, COL-II, COL-II, BMP2, and Runx2 through β-catenin in OPF rats. In conclusion, JPBS inhibits Axin/GSK-3β expression, activates the β-catenin signaling, and promotes the osteogenic differentiation of SMSCs.
骨质疏松症是一种代谢性骨病。β-连环蛋白与骨折相关。健脾补肾方（JPBS）可促进骨质疏松性骨折（OPF）的愈合。然而，JPBS通过β-连环蛋白介导骨骼肌卫星细胞（SMSCs）在OPF中的作用机制尚不清楚。分离SMSCs并将其分为五组。结果显示，孵育7天后，低、中、高剂量含JPBS血清组的SMSCs存活率显著更高。含JPBS血清干预组中碱性磷酸酶（ALP）活性及SMSCs矿化数量升高。构建Axin、糖原合成酶激酶-3β（GSK-3β）、β-连环蛋白的小干扰RNA（siRNAs）并转染至细胞中。siRNAs转染分别降低了Axin、GSK-3β和β-连环蛋白的表达。β-连环蛋白-siRNA逆转了ALP活性、SMSCs矿化数量以及β-连环蛋白、骨形态发生蛋白2（BMP2）、Runx2、I型胶原（COL-I）、SP7/成骨相关转录因子（Ostrix）、骨钙素和骨形态发生蛋白-7（BMP-7）的表达。转录组学结果表明，与OPF相关的肿瘤坏死因子（TNF）信号通路得到富集。通过切除卵巢构建OPF模型的SD大鼠。JPBS通过β-连环蛋白降低OPF大鼠中I型前胶原氨基端前肽（PINP）、ALP、I型胶原羧基端肽（CTX）和I型胶原氨基端肽（NTX）的水平，同时通过β-连环蛋白增加骨折断端处Runx2、β-连环蛋白的表达。此外，JPBS通过β-连环蛋白降低OPF大鼠的骨矿含量（BMC）、骨密度（BMD）和骨体积分数（BV/TV）并改善病理损伤。JPBS降低OPF大鼠中Axin、GSK-3β mRNA及蛋白的表达，但通过β-连环蛋白增加β-连环蛋白、配对盒基因7（Pax7）、II型胶原（COL-II）、BMP2和Runx2的表达。总之，JPBS抑制Axin/GSK-3β表达，激活β-连环蛋白信号通路，并促进SMSCs的成骨分化。
