Effect of TMED10 on TGF-β/Smads Signaling Pathway in Osteoarthritis
Objective:To investigate the effect of transmembrane p24 trafficking protein 10(TMED10)on transforming growth factor-β(TGF-β)/Smads signaling pathway in osteoarthritis.Methods:The rat chondrocytes were divided into Control group,IL-1βgroup,NC-sh+IL-1β group,TMED10-sh+IL-1β group,NC-OE+IL-1β group and TMED10-OE+IL-1β group.Control group and IL-1βgroup were not transfected,and NC-sh+IL-1β group,TMED10-sh+IL-1β group,NC-OE+IL-1β group and TMED10-OE+IL-1β group were transfected with NC-sh,TMED10-sh,NC-OE and TMED10-OE,respectively.After transfection,chondrocytes in all groups except Control group were treated with interleukin-1β(IL-1β)(10 ng/mL)for 24 h to simulate the pathological microenvironment of OA chon-drocytes.Cell proliferation was detected by MTT assay.Apoptosis was detected by TUNEL method.The OA model was established by anterior cruciate ligament amputation plus medial meniscectomy.The OA model rats were divided into OA group,OA+NC-sh group and OA+TMED10-sh group(n=12).The rats in Sham group(n=12)were undergoing sham operation.The rats in Sham group and OA group were injected 40 μL normal saline into the joint cavity.The rats in OA+NC-sh group and OA+TMED10-sh group were injected with 40μL NC-sh and TMED10-sh(titer 1 × 109 TU/mL),respectively.The injection was given once a week for 4 weeks.The shape of knee car-tilage was evaluated by saffranine O/solid green staining.Chondrocyte apoptosis was detected by TUNEL method.The mRNA level of TMED10 in chondrocytes and rat articular cartilage was detected by RT-qPCR.Protein expression levels of TMED10,TGF-β1,Smad2,p-Smad2,Smad3,Collagen Ⅱ and matrix metalloproteinase-13(MMP-13)in chondrocytes and rat articular cartilage were detected by Western blot.Results:Compared with Control group,the positive rate of TUNEL,the TMED 10 mRNA,TMED 10 and MMP-13 protein levels of IL-1β group increased(P＜0.05),the relative cell viability and the protein levels of TGF-β1,p-Smad2/Smad2,p-Smad3/Smad3 and Collagen Ⅱ of IL-1 β group decreased(P＜0.05).Compared with IL-1 β group and NC-sh+IL-1β group,the positive rate of TUNEL,the TMED 10 mRNA,TMED 10 and MMP-13 protein levels of TMEDl0-sh+IL-1β group decreased(P＜0.05),the relative cell viability and the protein levels of TGF-β1,p-Smad2/Smad2,p-Smad3/Smad3 and Collagen Ⅱ of TMED10-sh+IL-1β group increased(P＜0.05).Compared with IL-1 β group and NC-OE+IL-1β group,the positive rate of TUNEL,the TMED 10 mRNA,TMED 10 and MMP-13 pro-tein levels of TMED10-OE+IL-1β group decreased(P＜0.05),the relative cell viability and the protein levels of TGF-β1,p-Smad2/Smad2,p-Smad3/Smad3 and Collagen Ⅱ of TMED10-OE+IL-1β group increased(P＜0.05).Compared with Sham group,the OARSI score,the positive rate of TUNEL,the TMED10 mRNA,TMED10 and MMP-13 protein levels of OA group increased(P＜0.05),the protein levels of TGF-β1,p-Smad2/Smad2,p-Smad3/Smad3 and Collagen Ⅱ protein in OA group decreased(P＜0.05).Compared with OA group and OA+NC-sh group,the OARSI score,the positive rate of TUNEL,the TMED 10 mRNA,TMED 10 and MMP-13 pro-tein levels of OA+TMED10-sh group decreased(P＜0.05),and the protein levels of TGF-β1,p-Smad2/Smad2,p-Smad3/Smad3 and Col-lagen Ⅱ in OA+TMED10-sh group increased(P＜0.05).Conclusions:This study suggests that TMED 10 may lead to chondrocyte loss and cartilage degeneration by inhibiting TGF-β/Smads signaling pathway,and TMED10/TGF-β/Smads signaling pathway may be a new tar-get for the treatment of OA.
OsteoarthritisTransmembrane p24 trafficking protein 10Transforming growth factor-βChondrocyteTGF-β/Smads signaling pathway
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