Macrophage biospecific extraction and HPLC-ESI-MSⁿ analysis for screening immunological active components in Smilacis Glabrae Rhizoma

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• 作者：Zhao-Guang Zheng^a ; ^c ; Ting-Ting Duan^a ; ^c ; Bao He^c ; Dan Tang^a ; ^c ; Xiao-Bin Jia^b ; Ru-Shang Wang^c ; Jia-Xiao Zhu^c ; You-Hua Xu^a ; ^c ; Quan Zhu^a ; ^c ; ^{qzhu40@yahoo.com.cn} ; ^{qzhu40@163.com} ; Liang Feng^a ; ^b ; ^c ; ^{wenmoxiushi@163.com}
• 关键词：Macrophage biospecific extraction ; HPLC&ndash ; ESI-MS ; Rhizoma Smilacis Glabrae ; Immunological active component ; Macrophage migration and adhesion
• 刊名：Journal of Pharmaceutical and Biomedical Analysis
• 出版年：2013
• 出版时间：15 April, 2013
• 年：2013
• 卷：77
• 期：Complete
• 页码：44-48
• 全文大小：574 K

文摘

A cell-permeable membrane, as typified by Transwell insert Permeable Supports, permit accurate repeatable invasion assays, has been developed as a tool for screening immunological active components in Smilacis Glabrae Rhizoma (SGR). In this research, components in the water extract of SGR (ESGR) might conjugate with the receptors or other targets on macrophages which invaded Transwell inserts, and then the eluate which contained components biospecific binding to macrophages was identified by HPLC-ESI-MSⁿ analysis. Six compounds, which could interact with macrophages, were detected and identified. Among these compounds, taxifolin (2) and astilbin (4) were identified by comparing with the chromatography of standards, while the four others including 5-O-caffeoylshikimic acid (1), neoastilbin (3), neoisoastilbin (5) and isoastilbin (6), were elucidated by their structure clearage characterizations of tandem mass spectrometry. Then compound 1 was isolated and purified from SGR, along with 2 and 4, was applied to the macrophage migration and adhesion assay in HUVEC (Human Umbilical Vein Endothelial Cells) -macrophages co-incultured Transwell system for immunological activity assessment. The results showed that compounds 1, 2 and 4 with concentration of 5 ¦ÌM (H), 500 nM (M) and 50 nM (L) could remarkably inhibit the macrophage migration and adhesion (Vs AGEs (Advanced Glycation End Produces) group, 1-L, 2-H and 4-L groups: p < 0.05; other groups: p < 0.01). Moreover, 1 and 4 showed satisfactory dose-effect relationship. In conclusion, the application of macrophage biospecific extraction coupled with HPLC-ESI-MSⁿ analysis is a rapid, simple and reliable method for screening immunological active components from Traditional Chinese Medicine.