Effect of Astragalus polysaccharides on proliferation,apoptosis and autophagy of periodontal ligament fibroblasts by regulating AMPK-mTORC1 pathway
Objective To investigate the effects of Astragalus polysaccharide(APS)on proliferation,apoptosis,autophagy and AMPK/mTORC1 pathway in periodontal ligament fibroblasts(HPLF).Methods HPLF cells were primarily cultured;HPLF cells were identified by immunohistochemistry,and divided into the control group,0.1 mg·mL-1 group(0.1 mg·mL-1 APS),0.2 mg·mL-1 group(0.2 mg·mL-1 APS)and 0.4 mg·mL-1 group(0.4 mg·mL-1 APS).The cell counting Kit-8(CCK-8)method was used to detect the cell proliferation inhibition rate;Flow cytometry was used to detect the apoptosis rate;Autophagy of HPLF cells was observed by transmission electron microscope;Autophagy,apoptosis and expressions of pathway related proteins were detected by Western blot.Results Compared with the control group,the cell proliferation inhibition rate and apoptosis rate in 0.1 mg·mL-1 and 0.2 mg·mL-1 groups were significantly lower,compared with those in 0.2 mg·mL-1 group,the proliferation inhibition rate and apoptosis rate in 0.4 mg·mL-1 group were significantly higher.Compared with the control group,0.1 mg·mL-1 and 0.2 mg·mL-1 APS significantly inhibited autophagy,while 0.4 mg·mL-1 APS promoted autophagy;compared with those the control group,Beclin-1,LC3-Ⅱ/LC3-Ⅰ,cysteinyl aspartate specific proteinase(Caspase-3),bcl-2 associated X protein(Bax)and p-AMPK/AMPK in 0.1 mg·mL-1 and 0.2 mg·mL-1 groups were significantly lower,and the expression levels in 0.4 mg·mL-1 group were significantly higher than those in 0.2 mg·mL-1 group;The expression levels of p62,survivin,Bcl-2 and p-mTORC1/mTORC1 were opposite.Conclusion APS can promote the proliferation of HPLF cells and inhibit cell apoptosis,and may inhibit autophagy by regulating AMPK-mTORC1 pathway.
Astragalus polysaccharidesperiodontal ligament fibroblastsadenosine monophosphate protein kinase/mammalian target of rapamycin
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