Enrichment and purification process of astragalosides and their anti-human gastric cancer MKN-74 cell proliferation effect.

Yang Ouyang,Jianhua Huang,Zingting Ouyang,Jin Kang

doi:10.4314/ahs.v14i1.5

Abstract

Radix astragali mainly contains saponins, polysaccharides, flavonoids, amino acids and other chemical constituents of which total astragalosides have immunomodulatory, anti-viral, hepatoprotective, and gastric mucosa protective effects. To investigate the process conditions for extraction, purification and enrichment of total astragalosides by macroporous adsorption resin, and to study the inhibitory effect of total astragalosides on growth of human gastric cancer cell line MKN-74. UV spectrophotometry was applied to determine the adsorption and desorption capacity of macroporous adsorption resin on total astragaloside content, MTT assay was used to determine the inhibition of MKN-74 cell growth by total astragalosides. The dynamic adsorption performance of DA201 adsorption resin was examined, and the dynamic adsorption curve of total astragalosides on DA201 resin column was plotted. Meanwhile, eluent and elution flow rate were investigated, the results showed that the choice of eluent of 80% ethanol, and a flow rate of 5 BV/h could maximize the yield of total astragalosides. MTT assay found that astragalosides could relatively pronouncedly inhibit the proliferation of MKN-74 cells, and the inhibitory effect was enhanced with the increase of astragaloside dose and the extension of processing time, which showed a dose-and time-dependence. DA201 resin can effectively enrich total astragalosides, total astragalosides have an inhibitory effect on growth of MKN-74 cells.

Highlights

MaterialsRadix Astragali is the dried root of perennial herba- Reagents and drugs ceous plants Astragalus membranaceus (Fisch.) Bunge Radix Astragali extract, self-prepared, containing var. mongholicus (Bunge) Hsiao or A. membranaceus 51.37% of total astragalosides; human gastric cancer (Fisch.) Bunge of family Leguminosae.[1]
Pretreatment of resin Newly purchased DA201 and D101 macroporous resins were soaked in 2 column bed volumes (BV) of 95% ethanol for 24 h, respectively; after fully swollen, the resins were eluted with ethanol, and with distilled water until there was no smell of ethanol and set aside
Macroporous adsorption resin is a new type of highmolecular polymer with large pore size, and relatively large specific surface area.[8,9]

Summary

Introduction

Radix Astragali is the dried root of perennial herba- Reagents and drugs ceous plants Astragalus membranaceus (Fisch.) Bunge Radix Astragali extract, self-prepared, containing var. mongholicus (Bunge) Hsiao or A. membranaceus 51.37% of total astragalosides; human gastric cancer (Fisch.) Bunge of family Leguminosae.[1]. Radix Astragali is the dried root of perennial herba- Reagents and drugs ceous plants Astragalus membranaceus (Fisch.) Bunge Radix Astragali extract, self-prepared, containing var. Mongholicus (Bunge) Hsiao or A. membranaceus 51.37% of total astragalosides; human gastric cancer (Fisch.) Bunge of family Leguminosae.[1] It mainly con- cell line MKN-74, purchased from the Cell Bank of tains saponins, polysaccharides, flavonoids, amino acids Beijing Institute of Cell Biology, CAS. Protective, and gastric mucosa protective effects.[4,5] In this paper, process conditions for extraction, purifi- Instruments cation and enrichment of total astragalosides by ma- Model 680 microplate reader (Bio-Rad, USA); General croporous adsorption resin were investigated, and the TV-1221 UV-vis spectrophotometer (General, Beijing); inhibitory effect of total astragalosides on growth of CO2 incubator (model CO-150, NBS, USA); SW-CJ-SF human gastric cancer cell line MKN-74 was studied. DMSO, MTT and other chemical constituents,[2,3] of whichtotal astra- (Sigma, USA), fetal calf serum (Gibco), RPMI 1640 galosides have immunomodulatory, anti-viral, hepato- medium (Invitrogen). protective, and gastric mucosa protective effects.[4,5] In this paper, process conditions for extraction, purifi- Instruments cation and enrichment of total astragalosides by ma- Model 680 microplate reader (Bio-Rad, USA); General croporous adsorption resin were investigated, and the TV-1221 UV-vis spectrophotometer (General, Beijing); inhibitory effect of total astragalosides on growth of CO2 incubator (model CO-150, NBS, USA); SW-CJ-SF human gastric cancer cell line MKN-74 was studied. clean bench (Suzhou Purification Equipment Factory, Sujing Group)

Methods

Results

Discussion