Fractions of Shen-Sui-Tong-Zhi Formula Enhance Osteogenesis Via Activation of β-Catenin Signaling in Growth Plate Chondrocytes

Abstract

Background: Shen-sui-tong-zhi formula (SSTZF) has been used to treat osteoporosis for decades and shows excellent clinical efficacy. This article aims to explore the optimal anti-osteoporotic ingredient and its precise mechanisms in mice models.Methods: In this study, we first screened the optimal anti-osteoporosis fraction of SSTZF extract in vivo, and then further explored the mechanism of its effects both in vivo and in vitro. Ten-week-old female C57BL/6J mice were administrated with each fraction of SSTZF. At 10 weeks after ovariectomy (OVX), femurs were collected for tissue analyses, including histology, micro-CT, biomechanical tests, and immunohistochemistry for ALP, FABP4, and β-catenin. Additionally, we also evaluated the mRNA expression level of ALP and FABP4 and the protein expression level of β-catenin after being treated with SSTZF extract in C3H10T1/2 cells. Moreover, we investigated the anti-osteoporosis effect of SSTZF extract on mice with β-catenin conditional knockout in growth plate chondrocytes (β-cateninGli1ER mice) through μCT, histology, and immunohistochemistry analyzes.Results: At 10 weeks after treatment, osteoporosis-like phenotype were significantly ameliorated in SSTZF n-butanol extract (SSTZF-NB) group mice, as indicated by increased trabecular bone area and ALP content, and decreased lipid droplet area and FABP4 content. No such improvements were observed after being treated with other extracts, demonstrating that SSTZF-NB is the optimal anti-osteoporosis fraction. Additionally, the elevated β-catenin was revealed in both OVX mice and C3H10T1/2 cells with SSTZF-NB administered. Furthermore, a significant osteoporosis-like phenotype was observed in β-cateninGli1ER mice as expected. However, SSTZF-NB failed to rescue the deterioration in β-cateninGli1ER mice, no significant re-upregulated ALP and downregulated FABP4 were observed after being treated with SSTZF-NB, demonstrating that SSTZF-NB prevents bone loss mainly via β-catenin signaling.Conclusion: SSTZF-NB enhances osteogenesis mainly via activation of β-catenin signaling in growth plate chondrocytes. SSTZF-NB is the optimal anti-osteoporosis fraction of SSTZF and it can be considered a salutary alternative therapeutic option for osteoporosis.

Integration of Network Pharmacology and Experimental Validation to Explore the Pharmacological Mechanisms of Zhuanggu Busui Formula Against Osteoporosis

“…After fixation with paraformaldehyde, the femurs were scanned using micro-CT (mCT) equipment (Skyscan 1176, Bruker mCT N.V., Kontich, Belgium) at a voltage of 50 kV with a current of 500 mA and a resolution of 9 mm per pixel. The NRecon v1.6 and CTAn v1.15 software were utilized to reconstruct the threedimensional (3D) structure of the distal femoral metaphysis as we previously described (34).…”

“…Traditional Chinese medicine (TCM) herb formulae have their advantages and characteristics in fewer side-effects and early intervention (12,13). "Kidney dominating bone, generating marrow" is the theoretical basis of TCM in the treatment of OP, and the deficiency of kidney function leads to OP (14). Zhuanggu Busui formula (ZGBSF) is a novel TCM formula composed of Epimedium (Yin Yang Huo), Dipsaci Radix (Xu Duan), Paeoniae Radix Alba (Bai Shao), Chuanxiong Rhizoma (Chuan Xiong), Hedysarum Multijugum Maxim.…”

The complex pathophysiology of bone fragility in obesity and type 2 diabetes mellitus: therapeutic targets to promote osteogenesis

“…Studies in knock-out mice ( 113 ) and other aging studies ( 114 ) also showed that Runx2 deficiency resulted in impairment in osteoblastogenesis and depletion for satellite cells. Thus, it is likely that satellite cells and its gene machinery, play significant role on mediating the process of bone repair and thus, can be used as strategy in treatment ( 115 ). The next section discusses on the targets to minimize/nullify the inflammatory oxidative stress and enhance osteogenesis.…”

N-Butanol Extract of Modified You-Gui-Yin Attenuates Osteoclastogenesis and Ameliorates Osteoporosis by Inhibiting RANKL-Mediated NF-κB Signaling

“…The MYGY consists of ten herbs: Persicae semen, Cinnamomi cortex, Monkshood root, Eucommiae cortex, Lycii fructus, Dioscoreae rhizome, Carthami flos, Corni fructus, Glycyrrhizae radix and Preparata rehmanniae radix (Table 1), and has been widely applied to treat various chronic diseases with fewer side effects (12,13). Our recent study has shown that MYGY-based therapies achieve significant clinical outcomes in PMOP management (14). In addition, several herbs of MYGY have been reported to ameliorate osteoporosis by inhibiting osteoclast activity, such as Glycyrrhizae radix, Dioscoreae rhizoma, Corni fructus (12,15,16).…”