Effect and mechanism of astaxanthin in regulating tRF-VaLAAC on the biological functions of Chondrocytes in osteoarthritis
Objective:To explore the effect and mechanism of astaxanthin in regulating tRF-ValAAC on the biological functions of chondrocytes in osteoarthritis(OA).Methods:Chondrocytes were isolated from the articular cartilage of 10 OA patients who underwent total knee arthroplasty and 10 control patients with knee trauma but without OA at the Second People's Hospital of Changzhou from December 2021 to September 2023.The cells were extracted using trypsinization and designated as the OA group and the control group,respectively.Chondrocytes from OA patients were treated with 20 μmol/L astaxanthin and designated as the OA+AST group.In the OA group,chondrocytes were transfected with tRF-ValAAC mimics or inhibitors,while in the OA+AST group,chondrocytes were transfected with tRF-ValAAC-inhibitors.After treatment,cell proliferation was assessed using the CCK-8 assay,and apoptosis was measured by flow cytometry.Levels of interleukin-6(IL-6),interleukin-1β(IL-1β)and tumor necrosis factor α(TNF-α)were measured using enzyme-linked immunosorbent assay(ELISA).The expression of tRF-ValAAC was detected by quantitative PCR(qPCR),and Western blot was performed to evaluate oxidative stress-related enzymes[superoxide dismutase 1(SOD1),glutathione peroxidase 4(GPX4)]and extracellular matrix(ECM)related proteins[type Ⅱ collagen(COL2),aggrecan(ACAN),and matrix metalloproteinase-13(MMP-13)].Results:Compared with the control group,tRF-ValAAC expression in OA chondrocytes was significantly decreased(P<0.05),while astaxanthin treatment significantly upregulated its expression(P<0.05),and OA chondrocytes showed decreased proliferation and increased apoptosis(all P<0.05).Compared with the OA group,the OA+AST group presented increased proliferation and decreased apoptosis(all P<0.05).Transfection with tRF-ValAAC mimics in OA chondrocytes increased proliferation and decreased apoptosis(all P<0.05).Transfection with tRF-ValAAC inhibitor in astaxanthin-treated OA chondrocytes increased proliferation and reduced apoptosis(all P<0.05).IL-6,IL-1β and TNF-α levels were increased in the OA group compared to the control group,while these cytokine levels were significantly reduced in the OA+AST group(all P<0.05).Transfection with tRF-ValAAC mimics reduced the levels of IL-6,IL-1β,and TNF-α in OA chondrocytes(all P<0.05),whereas transfection with tRF-ValAAC inhibitors in astaxanthin-treated OA chondrocytes increased these cytokine levels(all P<0.05).Compared with the control group,the OA group presented increased MMP-13 protein levels and decreased SOD1,GPX4,COL2,and ACAN protein levels(all P<0.05).Compared with the OA group,the OA+AST group presented decreased MMP-13 protein levels and increased SOD1,GPX4,COL2,and ACAN protein levels(all P<0.05).Transfection with tRF-ValAAC mimics reduced MMP-13 levels and increased SOD1,GPX4,COL2,and ACAN levels in OA chondrocytes(all P<0.05).Transfection with tRF-ValAAC inhibitors in astaxanthin-treated OA chondrocytes led to increased MMP-13 levels and decreased SOD1,GPX4,COL2,and ACAN levels in OA chondrocytes(all P<0.05).Conclusions:Overexpression of tRF-ValAAC promotes chondrocyte proliferation and inhibits cell apoptosis in OA.Astaxanthin may alleviate chondrocyte damage in OA by increasing the tRF-ValAAC,suggesting a protective role in OA joint chondrocytes.
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