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Spectrum-effect Relatsionship of Extract from Semen Sojae Praeparatrm Fermented by Bacillus natto on Proliferative Activity Toward Rat Osteoblasts

??OBJECTIVE To establish the fingerprint of the extract from Semen Sojae Praeparatrm fermented by different preparation, to investigate the effects on the proliferation in rat osteoblasts, and to analyze the spectrum-effect relasionship. METHODS The fingerprint of alcohol extract from Semen Sojae Praeparatrm fermented by one functional bacteria for 6, 12, 18 h and 1-7 d(H6, H12, H18 and D1~D7) were established by HPLC, and the proliferation of each extract on rat osteoblasts was measured by MTT. The spectrum-effect relasionship was analyzed by grey relation analysis. Qualitative analysis was performed by HPLC-DAD-MS for analysis of major active components in alcoholic extract. RESULTS Alcoholic extract of Semen Sojae Praeparatrm fermented by different preparation was demonstrated to show similar proliferative activity at the concentration of 100 ??g??L^-1. Semen Sojae Praeparatrm fermented for 4 d led to the osteoblasts proliferation rate of 43.6%. On the basis of UV and mass spectral analysis,sixteen flavonoids were tentatively characterized.The glycitein had the highest correlation with pharmacodynamics, genistin, glycitin and daidzin also show higher correlation than the other chromatography peaks. The proliferation were promoted significantly after the treatment with glycitein, genistin, and daidzin (10^-9-10^-6 mol??L^-1) by the verification experiment. CONCLUSION Fermentation time of 4 d is more appropriate. The results of grey relational analysis and the activity in vitro of glycitein, genistin and daidzin mutually support each other, which confirms the scientificness of the spectral-effect relationship analysis in this study. The substance basis of anti-osteoporosis effect in the pure bacterial fermented soybean and natural fermented soybean are basically the same.