利用小鼠原代软骨细胞体外过表达模型探究核糖体蛋白S29在骨关节炎中的作用OA

Objective To investigate the role of ribosomal protein S29(Rps29)in osteoarthritis using an in vitro Rps29-overexpressing mouse primary chondrocyte model.Methods Rps29 expression in cartilage of meniscectomy(DMM)-induced osteoarthritis(OA)mice was detected by immunohistochemical staining.Through primary mouse chondrocyte isolation and culture,an Rps29-overexpressing model was established through cell transfection with control and Rps29-overexpressing vectors carrying the green fluorescence reporter gene.Expression of Rps29 mRNA and protein in the Rps29-overexpressing cell model was examined with reverse-transcription fluorescence quantitative PCR(RT-qPCR)and Western Blot.EdU incorporation was used to analyze Rps29-overexpressing chondrocyte proliferation.Immunofluorescence,RT-qPCR,and Western Blot were also utilized to measure expression levels of key molecules in OA,such as Collagen Ⅱ,Collagen Ⅹ,tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),nuclear factor-κB(NF-κB),and matrix metalloproteinase-13(MMP13).Results Rps29 expression in the knee joint of the DMM group was increased compared with the control group.Rps29 mRNA and protein expression were both significantly increased in Rps29-overexpressing primary chondrocytes compared with the control group.Decreased proliferation,decreased Collagen Ⅱ expression,and increased Collagen Ⅹ expression were detected in Rps29-overexpressing chondrocytes,andTNF-α,IL-6,and NF-κB expression was also significantly increased.Conclusions The molecular pathological changes of Rps29-overexpressing primary chondrocytes resembled an OA phenotype.Rps29 promoted OA by activating inflammatory responses and NF-κB signaling,providing new clues and a preliminary theoretical basis for understanding the role of Rps29 in OA.