ABSTRACT
Aim To investigate the effect of puerarin on proliferation, differentiation, and mineralization of MC3T3-E1 cells and the expression of TRPM3 mRNA.Methods Proliferation, differentiation, and mineralization of puerarin in MC3T3-E1 cells were determined using CCK-8 assay, alkaline phosphatase(ALP) activity assay, and Alizarin Red Staining, respectively.Effect of puerarin on cell cycle and intracellular calcium concentration of MC3T3-E1 cells was detected by flow cytometry.RT-PCR was used to detect the effect of puerarin on TRPM3 mRNA expression.Resuls 0.1, 1, 10 μmol·L-1 puerarin significantly promoted the proliferation of MC3T3-E1 cells, reduced the proportion of cells in G1 phase, increased the proportion of G2 and S phase, of which 0.1 μmol·L-1 concentration effect was the most significant.Compared with control group, the ALP activity and mineralized nodule area of 0.1 μmol·L-1 puerarin group were significantly increased.The expression of TRPM3 mRNA and the intracellular calcium concentration were significantly decreased in 0.1 μmol·L-1 puerarin group.Conclusion Puerarin can promote the proliferation, differentiation, and mineralization of MC3T3-E1 cells, and reduce the expression level of TRPM3 mRNA and intracellular calcium concentration.
ABSTRACT
Aim To study proliferation capacity of cell and the target relationship between microRNA and Runx2 after effect of puerarin on osteoblasts MC3 T3-E1 . Methods The proliferation capacity of cell was detected by MTT after effect of puerarin on osteoblasts MC3 T3-E1 . The vitality of osteoblasts was detected by activity of alkaline phosphatase. The expression level of mRNA and protein of Runx2 were detected by real-time quantitative PCR and Western blot. The result of miRNA expression spectrum was compared with the predicted result to determine the Runx2-targeting miR-NAs. The expression levels of miRNAs possiby targeted to Runx2 were detected by real-time quantitative PCR. The RhoE 3′UTR vector and RhoE mut 3′UTR vector were constructed. miRNA-204 mimics and miRNA-204 NC were synthetised. The target genes were verified by dual luciferase report gene assay. Results After osteo-blasts treated with puerarin, proliferation capacity and activity of cells were enhanced , expression levels of mRNA and protein of Runx2 were both increased , the expression levels of miRNA-204 and miRNA-344 f-5 p were declined, the expression levels of miRNA-2861 was increased,the expression levels of miRNA-23a-5p, miRNA-770-5 p and miRNA-871-5 p showed no obvious change. According to the results of dual luciferase re-porter gene method after cell transfection of 48 h, only set of 3′UTR Runx2+mimics the miRNA-204 of fluo-rescein protein expression level decreased significantly, showing only the miRNA-204 inhibits Runx2 3′UTR report gene expression. Conclusion Puerarin pro-motes the proliferation of osteoblasts and regulates the miRNAs which possibly target to Runx2 .
ABSTRACT
Flavonoids, which is composed of a C6-C3-C6 backbone, is a kind of secondary metabolites of Chinese materia medica (CMM). It has received extensive attention from all over the world and obtained a rapid develop-ment due to its widespread distribution, structural complexity and active diversity. This article has summarized the progress of flavonoids both in antioxidant activity and in mechanism, so as to provide some references for the re-search and development of new medicine of flavonoids with antioxidant activity.
ABSTRACT
With the modernization of traditional Chinese medicine (TCM), classic extraction methods such as sol-vent extraction, steam distillation, squeezing method, sublimation method have become much more difficult in sat-isfying needs of the development of the society as a result of low extraction rate of effective components, low per-centage impurity clearance. Therefore, in recent years the new extraction techniques with the advantages of green environmental protection , conditional stability , high extraction rate , strong biological activity , have been widely used in TCM extraction. These new extraction techniques have already shown each of its strength. The application progress on new extraction technology of Chinese medicine polysaccharide, both in standalone application and in collaboration application, were discussed in this article, so as to make new technologies be applied more reason-ably and effectively in Chinese medicine polysaccharide components.