Research On Extraction Process And Increase Bone Mineral Density Effect Of Luronggubao

Objectives: In order to develop a new kind of health-care food with fine processing that increases bone mineral density, this research focused on the extraction process and the effect of increasing bone mineral density of Chinese Medicine Luronggubao, which comprises of Cornu Cervi Pantotrichum, aceranthus sagittatus S.et Z, Cervus Nippon Temminck and Fructus Sophorae.Methods: 1. Extraction process research: orthogonal experiment L9(34) was used to optimize extraction process of Luronggubao. The content of icariin was determined by high-performance liquid chromatography(HPLC). The transfer rate of icariin was identified as the standard to inspect factors of water extraction of aceranthus sagittatus S.et Z and Fructus Sophorae. Factors of water extraction were water volume used to extraction, extraction frequency and extraction time. The total nitrogen content was determined by Kjeldahl and it was identified as the standard to inspect factors of water extraction of Cervus Nippon Temminck, determining the optimum extraction process. 2. Effective components and specific marker measurement: The content of total flavonoids of effective components in Luronggubao was determined by ultraviolet visible spectrophotometry. HPLC was used to detect the level of specific marker icariin. 3. Research on the effect on osteoblastic cell: The effect of Luronggubao on MC3T3-E1 proliferation and the protective effect of Luronggubao against H2O2 in MC3T3-E1 cell were determined by MTT assay. MC3T3-E1 differentiation was also detected. 4. Research on the effect on ovariectomised rats: Ovariectomised rats were fed on Luronggubao administration for 12 consecutive weeks and its impact on blood calcium, bone calcium, bone mineral density was detected. Bone mineral density was measured by Single Photon Absorptiometry, and Atomic Absorption Spectroscopy was used to measure bone calcium content.Results: 1. Extraction process research: The optimum extraction process for aceranthus sagittatus S.et Z and Fructus Sophorae: 10-fold amount of water, 3-time decoction, and 1.5 hours per decoction; the optimum extraction process for deer bone: 12-fold amount of water, 4-time decoction, 6 hours per decoction. 2. Effective components and specific marker: The total content of flavonoids was 21.08mg/g. Specific marker icariin content was 5.91mg/g. Data above indicated that this method is an acceptable and repeatable way for measuring effective components or specific marker. 3. Research on the effect on osteoblastic cell: MC3T3-E1 proliferation was promoted under exposure of Luronggubao, and the protective effect of Luronggubao against H2O2 injury in MC3T3-E1 cell was also observed. 4. Research on the effect on ovariectomised rats: Compared to the model group, Luronggubao administration increased the level of bone mineral density, bone calcium, blood calcium significantly.Conclusions: The Luronggubao prepared in this research boasts accessible process with remarkable repetition, explicit effective components, exclusive content detection methods and significant results of increasing bone mineral density in a safe and stable way.