Objective Osteogenesis is vitally important for bone defect repair,and Zuo Gui Wan(ZGW)is a classic prescription in traditional Chinese medicine(TCM)for strengthening bones.However,the specific mechanism by which ZGW ...Objective Osteogenesis is vitally important for bone defect repair,and Zuo Gui Wan(ZGW)is a classic prescription in traditional Chinese medicine(TCM)for strengthening bones.However,the specific mechanism by which ZGW regulates osteogenesis is still unclear.The current study is based on a network pharmacology analysis to explore the potential mechanism of ZGW in promoting osteogenesis.Methods A network pharmacology analysis followed by experimental validation was applied to explore the potential mechanisms of ZGW in promoting the osteogenesis of bone marrow mesenchymal stem cells(BMSCs).Results In total,487 no-repeat targets corresponding to the bioactive components of ZGW were screened,and 175 target genes in the intersection of ZGW and osteogenesis were obtained.And 28 core target genes were then obtained from a PPI network analysis.A GO functional enrichment analysis showed that the relevant biological processes mainly involve the cellular response to chemical stress,metal ions,and lipopolysaccharide.Additionally,KEGG pathway enrichment analysis revealed that multiple signaling pathways,including the phosphatidylinositol-3-kinase/protein kinase B(PI3K/AKT)signaling pathway,were associated with ZGW-promoted osteogensis.Further experimental validation showed that ZGW could increase alkaline phosphatase(ALP)activity as well as the mRNA and protein levels of ALP,osteocalcin(OCN),and runt related transcription factor 2(Runx 2).What’s more,Western blot analysis results showed that ZGW significantly increased the protein levels of p-PI3K and p-AKT,and the increases of these protein levels significantly receded after the addition of the PI3K inhibitor LY294002.Finally,the upregulated osteogenic-related indicators were also suppressed by the addition of LY294002.Conclusion ZGW promotes the osteogenesis of BMSCs via PI3K/AKT signaling pathway.展开更多
A rapid method based on rapid resolution liquid chromatography（RRLC） coupled with a diode array detector（DAD） was developed for the simultaneous determination of six major constituents（magnoflorine,jatrorrhizine,...A rapid method based on rapid resolution liquid chromatography（RRLC） coupled with a diode array detector（DAD） was developed for the simultaneous determination of six major constituents（magnoflorine,jatrorrhizine,coptisine,palmatine,berberine and evodiamine） in traditional Chinese medicine（TCM） Zuo Jin Pill（ZJP）.The satisfactory chromatographic separation was carried on an Eclipse Plus C18 column（1.8 μm i.d.,150 mm×4.6 mm） by linear gradient elution with a mobile phase of acetonitrile-acetate buffer.All the calibration curves show good linearity（r2 0.9998）.The detection limits and quantification limits ranged in 1.4―12 ng and 4.8―30 ng,respectively.The intra-and inter-day precisions were less than 0.63% with accuracies 98.60%―100.78%,and the recoveries were from 99.45% to 100.46%.Furthermore,hierarchical cluster analysis（HCA） was used to evaluate the variation of the herbal prescription.The results demonstrate that this analytical method is simple,sensitive and reliable for rapidly analyzing six major bioactive compounds in ZJPs and is helpful to comprehensively evaluating the quality of this TCM.展开更多
文摘Objective Osteogenesis is vitally important for bone defect repair,and Zuo Gui Wan(ZGW)is a classic prescription in traditional Chinese medicine(TCM)for strengthening bones.However,the specific mechanism by which ZGW regulates osteogenesis is still unclear.The current study is based on a network pharmacology analysis to explore the potential mechanism of ZGW in promoting osteogenesis.Methods A network pharmacology analysis followed by experimental validation was applied to explore the potential mechanisms of ZGW in promoting the osteogenesis of bone marrow mesenchymal stem cells(BMSCs).Results In total,487 no-repeat targets corresponding to the bioactive components of ZGW were screened,and 175 target genes in the intersection of ZGW and osteogenesis were obtained.And 28 core target genes were then obtained from a PPI network analysis.A GO functional enrichment analysis showed that the relevant biological processes mainly involve the cellular response to chemical stress,metal ions,and lipopolysaccharide.Additionally,KEGG pathway enrichment analysis revealed that multiple signaling pathways,including the phosphatidylinositol-3-kinase/protein kinase B(PI3K/AKT)signaling pathway,were associated with ZGW-promoted osteogensis.Further experimental validation showed that ZGW could increase alkaline phosphatase(ALP)activity as well as the mRNA and protein levels of ALP,osteocalcin(OCN),and runt related transcription factor 2(Runx 2).What’s more,Western blot analysis results showed that ZGW significantly increased the protein levels of p-PI3K and p-AKT,and the increases of these protein levels significantly receded after the addition of the PI3K inhibitor LY294002.Finally,the upregulated osteogenic-related indicators were also suppressed by the addition of LY294002.Conclusion ZGW promotes the osteogenesis of BMSCs via PI3K/AKT signaling pathway.
基金Supported by the National Natural Science Foundation of China(No.30725045)the Special Program for New Drug Innovation of the Ministry of Science and Technology,China(Nos.2009ZX09311-001 and 2008ZX09101-Z-029)+2 种基金China Postdoctoral Science Foundation(No.20080441304)Shanghai Leading Academic Discipline Project,China(No.B906)the Scientific Foundation of Shanghai City,China(Nos.07DZ19728,09DZ1975700 and 09DZ1971500)
文摘A rapid method based on rapid resolution liquid chromatography（RRLC） coupled with a diode array detector（DAD） was developed for the simultaneous determination of six major constituents（magnoflorine,jatrorrhizine,coptisine,palmatine,berberine and evodiamine） in traditional Chinese medicine（TCM） Zuo Jin Pill（ZJP）.The satisfactory chromatographic separation was carried on an Eclipse Plus C18 column（1.8 μm i.d.,150 mm×4.6 mm） by linear gradient elution with a mobile phase of acetonitrile-acetate buffer.All the calibration curves show good linearity（r2 0.9998）.The detection limits and quantification limits ranged in 1.4―12 ng and 4.8―30 ng,respectively.The intra-and inter-day precisions were less than 0.63% with accuracies 98.60%―100.78%,and the recoveries were from 99.45% to 100.46%.Furthermore,hierarchical cluster analysis（HCA） was used to evaluate the variation of the herbal prescription.The results demonstrate that this analytical method is simple,sensitive and reliable for rapidly analyzing six major bioactive compounds in ZJPs and is helpful to comprehensively evaluating the quality of this TCM.
