Sanguinarine Regulates The Proliferation And Apoptosis Of Osteoblasts Through AMPK Signaling Pathway

Objective:To study the mechanism of sanguinarine regulating osteoblast proliferation and apoptosis.Methods: 1.Primary osteoblasts were isolated from newborn rats.ALP staining was used to identify the osteoblasts.2.5 different concentrations of sanguinarine treated primary rat osteoblasts and MC3T3-E1 cells in mice,CCK-8 test,ALP activity experimental determination of sanguinarine on osteoblast proliferation promoting effect,The expression of AMPKa1 and P-AMPKa1 was detected by Western blot;Osteoblast group: control group,empty vector group,AMPKa1 interference group,high dose sanguinarine + empty vector group,AMPKa1 interference group load + high sanguinarine group;CCK-8 assay was used to detect cell proliferation,ALP activity and cell apoptosis by flow cytometry;Western blot detection of AMPKa1,P-AMPKa1,BMP2,OSX,OPG,Bcl2,Bax,Smad1,P-smad1 gene expression;Result:1.The inverted microscope separation of primary osteoblasts were spindle shaped,triangular or irregular polygon,and have extended number of projections are consistent bone cells typical of alkaline phosphatase staining showed that most cells in the cytoplasm to form granular or lamellar precipitate purple black,positive reaction and according to the biological characteristics of osteoblasts.2.sanguinarine can stimulate the activity of alkaline phosphatase increased the expression of AMPKa1 and promote the proliferation of osteoblasts,and exhibited a concentration dependent manner.3,sanguinarine can reverse AMPKa1 siRNA on proliferation of osteoblasts and promote accelerated apoptosis;increase the expression of P-AMPKa1,OSX,BMP2 can regulate OPG,Bcl2 and P-smad1 genes were up-regulated and down regulated expression of Bax gene.Conclusion:1,sanguinarine is by increasing the activation of AMPKa1,thereby promoting the activity of osteoblast proliferation,stimulation of alkaline phosphatase.And showed concentration dependent.2,sanguinarine adjusted to the proliferation and apoptosis of bone cells through the AMPK pathway,which is controlled by P-AMPKa1 BMP2,OSX,OPG,Bcl2,P-smad1 gene expression and down-regulation of Bax implementation.