Objective To investigate the impacts of sinomenine(SN)on interleukin-1β(IL-1β)induced autophagy and apoptosis in articular chondrocytes by regulating the adenosine monophosphate activated protein kinase(AMPK)/mam-malian target of rapamycin(mTOR)/UNC-51-like kinase 1(ULK1)signaling pathway.Methods Joint chondrocytes were grouped into control group(normal culture),IL-1βgroup(induced with 10 ng/mL IL-1β for 12 hours),L-SN,M-SN,H-SN groups(adding SNs of 25,50,and 100 μM on the basis of IL-1β induction),and SN+Compound C group(adding 10 μmol/L AMPK inhibitor Compound C pn the basis of the H-SN group).MTT assay,transmission electron microscopy(TEM),and flow cytometry were used to detect the effects of SN on proliferation,autophagy,and apoptosis of articular chondrocytes in each group.ELISA kit was applied to detect the expression of COX-2,TNF-α,MMP-3,and MMP-13 of cells in various groups.Western blotting(WB)was applied to detect the protein expression of p-AMPK,AMPK,p-mTOR,mTOR,p-ULK1,and ULK1 in the cells of each group.Results Compared with the Control group,the A490 values of articular chondro-cytes,the expression of p-AMPK/AMPK and p-ULK1/ULK1 proteins in the IL-1 β group were decreased,the autophagy vacuole count,apoptosis rate,the expression of COX-2,TNF-α,MMP-3,MMP-13,and p-mTOR/mTOR proteins were in-creased(P＜0.05).Compared with IL-1β group,the A490 value,number of autophagic vacuoles,the expression of p-AMPK/AMPK and p-ULK1/ULK1 proteins in the L-SN group,M-SN group,and H-SN group were increased,the apoptosis rate,the expression of COX-2,TNF-α,MMP-3,MMP-13,and p-mTOR/mTOR proteins were decreased(P＜0.05).Compared with H-SN group,the A490 value,number of autophagic vacuoles,the expression of p-AMPK/AMPK and p-ULK1/ULK1 proteins in the SN+Compound C group were decreased,the apoptosis rate,the expression of COX-2,TNF-α,MMP-3,MMP-13,and p-mTOR/mTOR proteins were increased(P＜0.05).Conclusion SN can promote IL-1β induced autophagy in articular chon-drocytes and inhibit cell apoptosis,which may be achieved by activating the AMPK/mTOR/ULK1 signaling pathway.
Joint chondrocytesSinomenineAdenosine monophosphate activated protein kinaseMammalian tar-get of rapamycinUNC-51 like kinase 1Interleukin-1βArticular chondrocyteAutophagyApoptosis
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