The Interventional Effects of Tubson-2 Decoction on Ovariectomized Rats as Determined by a Combination of Network Pharmacology and Metabolomics

Abstract

Post-menopausal osteoporosis (PMOP) is associated with estrogen deficiency and worldwide, is becoming increasingly more prevalent in aging women. Various anti-PMOP drugs have been developed to reduce the burden of PMOP; generally, these drugs are efficacious, but with some adverse side effects. Tubson-2 decoction (TBD), a popular traditional Mongolian medicine, has been used to treat PMOP for centuries. However, the precise mechanisms underlying the action of TBD on PMOP have yet to be fully elucidated. Herein, we combined network pharmacology with untargeted metabolomics to identify the key targets and metabolic pathways associated with the interventional effects of TBD on ovariectomized (OVX) rats. Furthermore, we investigated the bone histomorphometry of eight different groups of rats to evaluate the therapeutic effect of TBD. First, we established a TBD-target/PMOP network via network pharmacology; this network identified three key protein targets—vitamin D receptor (VDR), cytochrome P450 19A1 (CYP19A1), and 11β-hydroxysteroid dehydrogenase type 1 (HSD11B1). Morphological analysis showed that severe impairment of the bone micro-architecture in OVX rats could be improved by TBD administration. The TBD-treated rats had a significantly lower bone surface-to-tissue volume (BS/TV) and a significantly smaller trabecular separation (Tb·Sp.) ( P <0.05) than the OVX rats; in contrast, bone volume fraction (BVF), trabecular thickness (Tb·Th.), trabecular number (Tb·N.), and bone mineral density (BMD) were significantly higher in the TBD-treated rats ( P <0.05). Multivariate and univariate analysis showed that OVX resulted in significant alterations in the concentrations of 105 metabolites and 11 metabolic pathways (P<0.05); in addition, 26 potential biomarkers were identified to investigate the progression of PMOP. Network pharmacology showed that major alterations in vitamin B6 metabolism were associated with the VDR target. Next, we validated the three crucial targets (VDR [P<0.01], HSD11B1 [P<0.01], and CYP19A1 [P<0.05]) by enzyme-linked immunosorbent assays (ELISAs) and demonstrated that the levels of these targets were elevated in the OVX group but reduced in the TBD-treatment group. Collectively, our results suggest that the interventional effects of TBD on OVX rats are likely to be associated with the down regulation of VDR. Our findings enhance our molecular understanding of the interventional effects of TBD on PMOP and will allow us to develop further TBD studies.

Integrated Strategy of Network Pharmacological Prediction and Experimental Validation Elucidate Possible Mechanism of Bu-Yang Herbs in Treating Postmenopausal Osteoporosis via ESR1

“…Integrated “herbs-compounds-targets” network ( Supplementary Figure S2 ) and KEGG pathway analysis ( Figure 2 ) predicted that BYD might possess its anti-osteoporosis effect on PMOP via the activation of ESR1 and estrogen signaling pathway. Estrogen deficiency was characterized as the pivotal mechanism of PMOP ( Shen et al, 2020 ; Yang et al, 2020 ). Estrogen replacement therapy had viewed as an effective but unsafe therapy for its severe side effects of up-regulating the potential of cancer ( Rachner et al, 2011 ).…”

Application of metabolomics in osteoporosis research

“…Network pharmacology suggested that major changes in vitamin B6 metabolism were related to vitamin D receptor targets. Thus, Tween-2 decoction on postmenopausal OP rats might be related to down-regulation of vitamin D receptor ( 142 ).…”

Reveal the Antimigraine Mechanism of Chuanxiong Rhizoma and Cyperi Rhizoma Based on the Integrated Analysis of Metabolomics and Network Pharmacology

“…There is growing interests in the network pharmacology, which has an advantage in elucidating the potential molecular mechanisms based on a drug–disease–target network ( Li et al, 2019 ). Recently, the integrated analysis of metabolomics and network pharmacology has been applied to elucidate the protein–metabolite interactions, which brings great inspiration to the mechanism research ( Wei et al, 2020 ; Yang et al, 2020 ; Zhang et al, 2020 ).…”