OM-MSCs Experimental Study Of Ages-induced Osteoarthritis Through Cx43 Mediated TLR4/MyD88/NF-κB Pathway

OBJECTIVE : To explore the role and molecular mechanism of connexin 43(CX43)in advanced glycation end products(AGEs)promoting osteoarthritis(OA),and to explore the possible mechanism of olfactory mucosal mesenchymal stem cells(OM-MSCs)in the treatment of OA.To provide theoretical basis and basis for finding effective new methods.MATERIALS AND METHODS :(1)To collect cartilage tissue samples from normal and OA subjects,Western Blot、RT-q PCR detection of Cx43 expression;(2)When human articular chondrocytes were treated with AGEs(200μg/ml),Western Blot、RT-q PCR detection of intracellular COL-Ⅱ、ACAN、IL-1β、MMP-13、ADAMTS-5、Cx43 expression;(3)After si-Cx43 treatment of human articular chondrocytes,Western Blot、RT-q PCR detection of intracellular IL-1β、MMP-13、TLR4、MyD88、NF-κB expression;(4)The AGEs induced OA chondrocytes were co-cultured with OM-MSCs using a Transwell device,To Western Blot、RT-q PCR the level of COL-Ⅱ、ACAN、IL-1β、MMP-13、ADAMTS-5、Cx43 、 TLR4 、 MyD88 、 NF-κB in the cell,And through CCK-8experiments to observe and detect the activity of human articular chondrocytes,Flow cytometry was used to observe and detect the apoptosis of human articular chondrocytes,The morphology of articular chondrocytes and the formation of autophagy lysosomes were observed by transmission electron microscope.RESULTS:(1)Compared with normal cartilage tissue,OA high expression of Cx43 in cartilage tissue(P<0.05);(2)AGEs treatment of human articular chondrocytes,The expression of COL-Ⅱ 、 ACAN decreased significantly(P<0.05),IL-1β 、 MMP-13 、 ADAMTS-5expression increased significantly(P<0.05);(3)AGEs treatment of human articular chondrocytes,Cx43 expression increased significantly(P<0.05);(4)si-Cx43 pretreatment of human articular chondrocytes,Treatment of human articular chondrocytes with AGEs h,24 Compare AGEs groups,IL-1β、MMP-13、Cx43、TLR4、MyD88、NF-κB expression decreased significantly(P<0.05);AGEs induced OA model cells were co-cultured with OM-MSCs,OM-MSCs significantly reduced the increase in IL-1β、MMP-13、ADAMTS-5、Cx43、TLR4、MyD88、NF-κB expression induced by AGEs(P<0.05),The expression of COL-Ⅱ、ACAN was increased(P<0.05);AGEs induced OA model cells were co-cultured with OM-MSCs,OM-MSCs significantly increased model cell activity(P<0.05),promoting its autophagosome production,And reduce its apoptosis rate(P<0.05).CONCLUSIONS:(1)Increasing Cx43 expression in knee cartilage tissue in OA patients;(2)AGEs can induce increased Cx43 expression in human chondrocytes,And to promote the increased IL-1β 、 MMP-13 expression in the chondrocytes through the activation of the TLR4/MyD88/NF‐κB signaling pathway;(3)OM-MSCs can inhibit the increase of Cx43 expression and the activation of the TLR4/MyD88/NF‐κB signaling pathway caused by AGEs;(4)After indirect coculture of model cells with OM-MSCs,Can mitigate the model cell inflammatory response,Enhance cell activity,Promote chondroblastic autophagy,And to reduce its apoptosis.