Preparative Purification of Bioactive Compounds from Flos Chrysanthemi Indici and Evaluation of Its Antiosteoporosis Effect.

Jia Li,Jiansheng Su,Jiquan Zhang,Weicai Liu,Xiaohui Mi,Xiaosheng Lin,Yuping Zhang

doi:10.1155/2016/2587201

Abstract

To understand the material basis and underlying molecular machinery of antiosteoporosis activity of the Flos Chrysanthemi Indici (FCI), the consequences of ethanol extract on the bone loss in mice induced due to ovariectomy (OVX) was evaluated. Also, the antiosteoporosis fraction obtained from the FCI ethanol extract was isolated and purified using a preparative high-speed countercurrent chromatography (HSCCC). The in vitro impact of the compounds was investigated on osteoblast proliferation and differentiation. The results revealed that ethyl acetate fraction with robust in vivo antiosteoporosis activity was obtained. The important compounds purified by HSCCC using gradient elution system included acacetin, apigenin, luteolin, and linarin. The four compounds enhanced the differentiation and proliferation of osteoblasts in MC3T3-E1 cells. They also augmented the mRNA levels of runt-related transcription factor 2 (Runx2), osteocalcin (OCN), osteopontin (OPN), and type I collagen (COL I). The AKT signaling pathway was also activated in MC3T3-E1 cells by the four compounds. The present study demonstrated that the antiosteoporosis effects of FCI did not depend on a single component, and HSCCC efficiently isolated and purified the antiosteoporosis bioactive compounds from FCI.

Highlights

Osteoporosis is characterized as a metabolic bone disease, wherein the bone microarchitecture is deteriorated due to disruption of bone formation and resorption [1, 2]
We reported that linarin extracted from Flos Chrysanthemi Indici (FCI) enhanced the osteoblast differentiation in MC3T3-E1 cells in a dosedependent manner [11]
As best known, we demonstrate for the first time that acacetin, apigenin, and luteolin extracted from FCI upregulated the gene expression of COL-I, OPN, and OCN, thereby promoting osteoblast differentiation

Summary

Introduction

Osteoporosis is characterized as a metabolic bone disease, wherein the bone microarchitecture is deteriorated due to disruption of bone formation and resorption [1, 2]. Hormone replacement therapy prevents osteoporosis or positively affects the bone formation during the treatment of the disease [5, 6]. Our previous study demonstrated that FCI extract treatment can effectively prevent ovariectomy- (OVX-) induced bone. Identification and purification of the bioactive fractions from FCI, for the treatment of osteoporosis, are vital for further investigation of its pharmacological mechanism. HSCCC could serve as an excellent method for rapid separation and purification of potentially applicative bioactive products from FCI. The present study describes the effect of the bioactive fractions from the ethanol extract of FCI on the bone loss induced by OVX in mice. A systematic evaluation of the ethanol extract of FCI against osteoporosis and HSCCC separation of antiosteoporosis active compounds has not been described

Materials and Methods

Solvent volume ratio

Results and Discussion

Conclusions