Spectral-effect relationship of immune regulation of deer bone extract based upon Grey correlation degree
OBJECTIVE To explore the correlation between fingerprint spectrum of deer bone and its immunomodulatory function and identify the active components contributing to its immunomodulatory effect.METHODS Ten groups of deer bone samples were prepared at different extraction temperatures and their fingerprint spectra detected.Multivariate statistical analysis was performed for common peaks in fingerprint spectra.Liquid chromatography-mass spectrometry(LC-MS)and standard prod-uct comparisons were employed for identification.Immunomodulatory function of deer bone was evaluated by measuring cellular viability and nitric oxide(NO)concentration of RAW264.7 injury model with ten groups of deer bone extracts.Spectrum-effect relationship of deer bone was established through grey relational analysis.RESULTS The fingerprint spectrum of deer bone had ten common peaks.The structures of these ten peaks were identified by LC-MS and six compounds detected through standard product comparisons.The similarity of deer bone fingerprint spectrum was greater than 0.9.Cluster analysis and principal compo-nent analysis yielded the same results,classifying deer bone into six categories.Orthogonal partial least squares discriminant analy-sis(OPLS-DA)revealed five chromatographic peaks with variable importance in projection(VIP)greater than 1,namely peaks 4,7(uridine),10(guanosine),5 and 9(inosine).Grey relational analysis indicated that all ten chromatographic peaks had a cor-relation greater than 0.6,suggesting that deer bone might exert its immunomodulatory function through multiple components.Nine chromatographic peaks had a correlation greater than 0.7,indicating a strong correlation.The first six chromatographic peaks were 7(uridine),10(guanosine),9(inosine),2(cytidine),3(uracil)and 6(hypoxanthine).In summary,three nucleo-side components of uridine,guanosine and inosine might be the major immunomodulatory components in deer bone.CONCLUSION Immunomodulatory function of deer bone may be the result of synergistic action of multiple nucleoside compo-nents.And uridine,guanosine and inosine are probably the major active components contributing to immunomodulatory effect of deer bone.
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