An MC3T3-E1 Cell Line Biomembrane Extraction and HPLC–ESI-MS n Method for Simultaneous Analysis of Potential Anti-Osteoporosis Components of Epimedium koreanum

Abstract

Aerial parts of Epimedium koreanum Nakai have been used as Herba Epimedii in China. Its anti-osteoporosis effect has attracted much attention in recent years. In this study, a method involving osteoblastic cell (MC3T3-E1 cell line) extraction and high-performance liquid chromatography-electrospray ionisation-mass spectrometry (HPLC-ESI-MS n ) was developed for screening of potential anti-osteoporosis agents in E. koreanum. Four compounds identified as epimedin A, epimedin B, epimedin C and icariin were found to interact with MC3T3-E1 cells and possessed potent osteoblast-stimulating activity as evaluated by cell proliferation [3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay] and differentiation [alkaline phosphatase (ALP) activity and Ca content] in vitro. The results suggest that these four flavonoids are the anti-osteoporosis constituents of Herba Epimedii and that the method combining MC3T3-E1 cell extraction with HPLC-ESI-MS n is rapid and effective in screening anti-osteoporosis agents from traditional Chinese medicines.

Chinese single herbs and active ingredients for postmenopausal osteoporosis: From preclinical evidence to action mechanism

“…Total flavones of Epimedium (TFE) inhibited reduction of procollagen type I N-terminal propeptide (PINP), and increased serum osteocalcin and type I collagen in OVX rats (13,14). Icariin, one of the major components of Herba Epimedium, decreased activities of serum tartrate-resistant acid phosphatase (TRAP) and bioactive components could stimulate the proliferation, differentiation and mineralization of osteoblasts (12,(28)(29)(30)(31)(32)(33)(34)(35)(36)(37)(38)(39)(40), suppress the adipogenesis of bone marrow-derived mesenchymal stem cells (BMSCs) (41)(42)(43)(44), inhibit the proliferation and differentiation of osteoclasts (45)(46)(47), and induce apoptosis and cell cycle arrest and suppress bone resorption of osteoclasts (48). In addition, icariin was found to significantly attenuate oxidative stress and apoptosis and preserve viability and osteogenic potential of osteoblasts exposed to hypoxia, which indicated that its anti-osteoporotic effect might be attributed to its anti-hypoxic activity (49).…”

Application progress of immobilized biomembrane in the discovery of active compounds of natural products

“…For example, HaCaT, HUVEC, and RAW 264.7 cell lines were chosen as the targets to screen the bioactive compounds from Fuzhengzhiyang decoction, and 14 types of potential active compounds were identified (Chen et al, 2020). Based on the specific binding reaction of osteoblast lines, the active ingredients have been successfully identified from Epimedium koreanum (Wang et al, 2012), Herba Epimedii (Jin et al, 2013), and Er‐Xian decoction (Wang, Xu, Wang, et al, 2019). Although living cells have a satisfactory number of targets, the isolation of living cells from NP extracts and preservation of their biological activity as much as possible are some of the challenges of this method.…”

Natural Products from Chinese Medicines with Potential Benefits to Bone Health

“…Besides icariin, a number of flavonol constituents of Epimedium are also osteo-active. Thus, epimedins A ( 40 , Figure 1), B ( 38 , Figure 1), C ( 39 , Figure 1) and icariin were demonstrated to interact with MC3T3-E1 cells in a biomembrane extraction model [211], and epimedin B and epimedin C promoted the proliferation of UMR-106 cells [178]. In a zebrafish model of anti-osteoporosis, epimedin A and baohuoside-1 (icarisinde II) ( 41 , Figure 1) were found to be active [212].…”