ABSTRACT
ObjectiveMicroRNAs (miRNA) play an important role in the development and regression of osteoporosis. This study aims to screen for miRNAs and genes closely related to osteoporosis, and to complete the construction of a miRNA-mRNA regulatory network of osteoporosis.MethodsThe gene chip expression profile of osteoporosis was obtained through the GEO database, and the differentially expressed genes (DEGs) and miRNAs were analyzed and screened via GEO2R. We used volcanic maps to display differential genes and miRNAs, and completed the GO and KEGG pathway analysis through the David online database. The String online database is used to complete PPI protein network analysis. The TargetScan, miRTarBase and miRDB were used to predict the targeted genes. Finally, the PPI and miRNA-mRNA regulatory networks were visualized by Cytoscape software.ResultsWe obtained 15 differential miRNAs and 174 differentially expressed genes through screening. The GO enrichment analysis mainly focused on drug response, angiogenesis, ion transport, regulation of small GTPase mediated signal ransduction, adaptive immune response, etc. NF-κB signaling pathway and HIF-1 signal were obtained through KEGG enrichment analysis. We obtained 10 hub genes through the cytohubba plug-in. We also obtained 3065 targeted genes by processing of seven miRNAs, and then intersected them with 174 DEGs to obtain 44 intersection genes. Finally, we successfully constructed a regulation map of miRNA-mRNA network. The miR-194-5p is significantly up-regulated in osteoporosis.ConclusionThe miR-194-5p might play an important role in osteoporosis by regulating its target gene CDH2, which provides candidate targets for the diagnosis and treatment of osteoporosis.
ABSTRACT
Objective WNT signaling pathway plays an important role in the formation, differentiation and maturation of bone cells, it is a classical intracellular signaling pathway involved in bone metabolism. DKK1 and Sost play a negative regulatory role in regulating bone mass and osteoblast differentiation, and are negative regulators of WNT signaling pathway. Estrogen-related receptor alpha (ERRα) regulates the functional activity of osteoblasts. The aim of study was to investigate the effect of ERRα on the transfection of MG63 cells and related proteins by the WNT signaling pathway inhibitor Dickkopf (DKK)1 and sclerostin (SOST) adenovirus vectors.Methods The cultured MG63 cells were divided into blank control group, silencing DKK1 group, silencing SOST group, silencing (DKK1+SOST) group, ERRα intervention empty adenovirus group, ERRα intervention silencing DKK1 group, ERRα intervention silencing SOST group, ERRα intervention silencing (DKK1+SOST) group. MG63 cells were transfected with packaged silencing DKK1 and SOST adenovirus vectors according to different groups. The activity of MG63 cells was detected by MTT assay, the activity of ALP was detected by alkaline phosphatase kit, and the concentration of calcium ion was analyzed by flow cytometry. Western blot was used to detect the expressions of low density lipoprotein associated protein 5 (LRP5), bone morphogenetic protein 2 (BMP2), osteopontin (OPN), osteoprotegerin(OPG).Results (1) Compared with blank control group, silencing DKK1, SOST, DKK1+SOST group and ERRα overexpression in the empty adenovirus group could increase cell activity, ALP activity, and decrease calcium ion concentration and increase the expressions of LRP5, BMP2, OPN, and OPG. Differences between groups were statistically significant(P0.05).Conclusion ERRα Overexpression can increase the activity of MG63 cells, ALP activity, LRP5, BMP2, OPN, and OPG proteins, and decrease the calcium ion concentration in silencing DKK1 and SOST adenovirus-transfected cells.
ABSTRACT
BACKGROUND: Chinese herbs have been shown to affect osteoblasts through Wnt/β-catenin signaling pathway. Sclerostin (Sost) is an inhibitor of Wnt signaling pathway and an inhibitor of osteogenesis, and it is also an antagonistic factor that regulates the differentiation and proliferation of osteoblasts. Bushen Jianpi Huoxue decoction (BJHD) is the empirical formula in the prevention and treatment of osteoporosis at the Affiliated Orthopedics Hospital, Guangzhou University of Chinese Medicine. BJHD can promote osteogenesis and effectively treat osteoporosis. OBJECTIVE: To observe the effect of BJHD on the cell proliferation, alkaline phosphatase activity and expression levels of related proteins in Sost-overexpressed adenovirus transfected osteoblasts. METHODS: The osteoblasts were divided into two groups: blank control group (empty adenovirus transfection) and Sost overexpressed group (Sost overexpression and adenovirus transfection). Two groups were cultured with blank serum and medicine serum. The cell proliferation was tested by cell counting kit-8 assay, and alkaline phosphatase activity was tested by alkaline phosphatase kit. Expression levels of bone regulation-related proteins were tested by western blot assay. RESULTS AND CONCLUSION: Compared with the blank serum, medicine serum could increase the cell viability at 24, 48 and 72 hours in both groups (P ＜ 0.01). Compared with the blank serum, medicine serum increased the alkaline phosphatase activity (P ＜ 0.01). Compared with the blank serum, medicine serum in the blank control group decreased the expression of low-density lipoprotein receptor-related protein 5 and osteoprotegerin (P ＜ 0.01), but showed no effect on the expression of osteopontin and tumor necrosis factor α. Medicine serum in the Sost-overexpressed group decreased the expression of low-density lipoprotein receptor-related protein 5, osteoprotegerin and osteopontin, increased the expression of tumor necrosis factor α (P ＜ 0.01). These results indicate that BJHD can increase the proliferation of recombinant adenovirus transfected Sost-overexpressed osteoblasts and alkaline phosphatase activity, and regulate the expression of low-density lipoprotein receptor-related protein 5, osteoprotegerin, osteopontin, and tumor necrosis factor α.
ABSTRACT
Hedgehog signaling pathway is a conserved and important signaling pathway involved in proliferation and differentiation of many types of cells. Latest studies have found that Hedgehog signaling pathway may induce MSCs osteoblast differentiation by increasing the expression of the Runx2 and Osx and inhibit MSCs differentiate to adipocyte. Hedgehog signaling pathway may also promote osteoblast proliferation by regulating cyclin. This review summarizes the mechanism that Hedgehog signaling pathway regulates osteoblast differentiation and proliferation,and concludes that Hedgehog signaling pathway can regulate bone metabolism. It might provide new ideas for the treatment of osteoporosis.
Subject(s)
Humans , Cell Differentiation , Core Binding Factor Alpha 1 Subunit , Genetics , Physiology , Hedgehog Proteins , Physiology , Mesenchymal Stem Cells , Cell Biology , Osteoblasts , Cell Biology , Osteoporosis , Drug Therapy , Signal Transduction , Physiology , Sp7 Transcription Factor , Transcription Factors , Genetics , PhysiologyABSTRACT
<p><b>OBJECTIVE</b>To investigate the effects of Gukang on bone-source alkaline phosphatase (BALP) and insulin-like growth factor 1 (IGF-1) in serum of spaying rats and the mechanism of curative effect of Gukang on osteoporosis.</p><p><b>METHODS</b>Sixty-eight 6-month-old SD rats were chosen and randomly divided into blank control group (22 rats with sham operation) and operation group (46 rats with spaying operation). Three months after operation, 10 rats were randomly chosen from each group and tested with bone mineral density in order to determine models of osteoporosis made. After modeling, operation group was divided into 3 sub-groups: operation model group, estrogen group and Gukang group, 12 rars in each group. Twelve rats remained in blank control group. Every group were treated through intragastric administration therapy (volume 10 ml/kg). Blank control group and operation model group were irrigated with distilled water,estrogen group with estrogen and Gukang group with Gukang. Three months after treatment, serum of all groups were collected and tested for E2, BALP and IGF-1 with ELISA.</p><p><b>RESULTS</b>The concentration of serum E2, BALP in estrogen group and Gukang group were higher than operation model group, there were significant difference (P < 0.05), but no significant difference in serum E2 between estrogen group and Gukang group (P > 0.05). The concentration of serum IGF-1 in Gukang group was higher than operation model group and blank control group, there were significant difference (P < 0.05).</p><p><b>CONCLUSION</b>Gukang can increase the level of E2, BALP and IGF-1 in serum of spaying rats. Thus, it can indirectly promote reproduction of osteoblasts, inhibit activity of osteoclasts and promote bone formation.</p>