关键词: TGFβ1基因, “健肾壮骨汤”, Wnt3a/β-catenin信号通路

Abstract: Objective To investigate the mechanism and effect of TGFβ1-mediated Jianshen Zhuanggu Tang （JSZGT） in regulating the Wnt3a/β-catenin signaling pathway on osteoblasts. Methods Osteoblast cell lines C518 and MC3T3-E1 were co-incubated with JSZGT and DKK1 （an inhibitor of the Wnt3a/β-catenin signaling pathway）. The differences in mRNA and protein levels of Wnt3a, TGFβ1, OPG, and RANKL in the cells were detected, and rescue experiments and RNA-seq analysis were conducted. Results After treating C518 and MC3T3-E1 cells with DKK1, the expression levels of TGFβ1, OPG, and RANKL were significantly downregulated （P<0.05）. DKK1 significantly reduced the mRNA and protein levels of Wnt3a, TGFβ1, OPG, and RANKL in both the sh TGFβ1 group and the TGFβ1 overexpression （TGFβ1OE） group （P<0.05）. In the C518 cell model, the knocked down expression of OPG gene inhibited the promoting effect of TGFβ1 on OPG expression; however, JSZGT treatment could restore this promoting effect of TGFβ1 on OPG expression. RNA-Seq results showed that TGFβ1, OPG, and RANKL were significantly differentially expressed genes. Conclusion The TGFβ1 gene mediates JSZGT to regulate the Wnt3a/β-catenin signaling pathway and thus modulate the expression of OPG/RANKL. The TGFβ1 gene may be an important target for the treatment of osteoporosis.

Key words: TGFβ1, Jianshen Zhuanggu Tang, Wnt3a/β-catenin signaling pathway