| Traditional Chinese medicine believe that osteoporosis belongs to “heumatism,bone atrophy”,Kidney deficiency is the main etiology and pathogenesis.“Kidney stores vital essence”“Kidney mains bone marrow”“Kidney mains bone” as the theoretical basis clinically,traditional Chinese medicine that reinforce kidney and strengthen bone are used as therapy.It was found out that the polysaccharides of Achyranthes bidentata(AB),Curculigo orchioides(CO),Cibotium barometz(CB)and Humulus lupulus Linn(HL),which were the used clinically,had positive activity on anti-osteoporosis.Four polysaccharides of CO were used to study their remedies on osteoporosis of ovariectomized rats.The serum of ovariectomized rats was processed to metabolism analysis and to find the biomarkers.Also the anti-osteoporosis effects of four AB polysaccharides were evaluated based on the biomarkers.Five puried polysaccharides of CB and HL were applied to research their effects on the proliferation,the differentiation and the mineralization of MC3T3-E1 cells.And the mechaniams of pro-osteogenesis were studied preliminarily from the genes and proteins level.The polysaccharides of Curculigo orchioides Gaertn(CO)that including CO50,CO70,CO90 and COB were applied to ovariectomized rats to observe the remedial effect of osteoporosis.Bone histomorphometry was used to analyze the left tibia section after toluidine blue staining.The standard of assessment of anti-osteoporosis was the changes of trabecula and cortical bones.The results indicated that CO50,CO70 and COB couldimprove osteoporosis in different grade.The three polysaccharides increased number and thickness of trabecular bone,percentage and the minimum width of cortical bone,percent labeled perimeter of cortical bone significantly.The mineral apposition rate and bone formation rate of cortical bone were increased obviously.The separation of bone trabecula was decreased.The results above indicated that CO50,CO70 and COB had apparent effects on remedying osteoporosis,and the CO70 had the best anti-osteoporosis effects.CBB-1,CBB-2,CBB-3,HL70-2 and HL90-1 were the purified polysaccharides of Cibotium barometz(CB)and Humulus lupulus Linn(HL),and they were applied to discuss the osteogenesis effects on mouse osteoblast-like MC3T3-E1 cells.The proliferation was tested with CCK-8,and activity of ALP was the indicator to evaluate the ability of differentiation of cells.Alizarin red S and cetylpyridinium were evaluated the effects of polysaccharides on mineralization of cells qualitatively and quantitatively.Realtime fluorescence quantitative PCR were used to test the change of expression of related genes.The results showed that CBB-1,HL70-2 and HL90-1could promoted the proliferation of MC3T3-E1;CBB-1,HL90-1 and CBB-2 increased the activity of ALP in cells;CBB-1 and HL90-1 increased the mineralization rate of cells significantly.CBB-1 and HL90-1promoted the differentiation and mineralization of cells.And CBB-1 up-regulated the expression of Ost and Runx2,which were the key genes in early stage of differentiation,and Bsp and Ocn,which were the key genes in later stage of differentiation.HL90-1up-regulated the expression of gene Runx2 and Bsp and protein BMP2 and Runx2 in early stage obviously and then promoted the differentiation and mineralization of cells.The serum of ovariectomized rats were processed to metabolism analysis to find the biomarkers of osteoporosis and evaluate the effects of polysaccharides of Achyranthes bidentata(AB)on osteoporosis.The content of glutarylcarnitine,lysoPC(18:1)and9-cis-retinoic acid had significant difference compared with the model group,then they were identified as the biomarkers.There were 27 compunds that had significant difference between the model group and the Sham group.15 compounds of 27 were related with lipid metabolism,which showed that osteoporosis had obvious relationship with dyslipidemia.The metabolism analysis results suggested that AB50 and AB90 had good anti-osteoporosisbioactivity.And AB50 had better effects,which was identical with the results in vivo. |
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