P300 Inhibits Intervertebral Disc Degeneration through Nrf2/HO-1/NF-κB Signaling in Rat Scoliosis Model
Objective To investigate the effect and potential regulatory mechanism of histone acetyltransferase P300 on the degeneration of nucleus pulposus cells(NPCs)in the intervertebral discs in the rat scoliosis model.Methods NPCs were cultured and divided into 4 groups:the un-treated control group,the interleukin-1 β(IL-1 β)-induced degeneration group(IL-1 β group),the IL-1 β combined with P300 treatment group(IL-1 β+P300 group),and the P300 treatment a-lone group(P300 group).Cell proliferation activity was detected using the Cell Counting Kit-8(CCK-8)assay.The levels of TNF-α and IL-6 were measured using enzyme-linked immunosorbent assay(ELISA).Cell apoptosis was determined using flow cytometry.The expression levels of sex de-termining region Y-box 9(SOX9)、collagen type Ⅱ(COL-Ⅱ),matrix metalloproteinase-13(MMP-13),A disintegrin and metalloproteinase with thrombospondin motifs 5(ADAMTS-5),in-hibitor of nuclear factor-kappa B-α(IKBα),phosphorylated IκBα(p-IKBα),phosphorylated NF-κB P65(p-P65),nuclear factor erythroid 2-related factor 2(Nrf2),and heme oxygenase-1(HO-1)in the cells were analyzed using Western blotting.Additionally,40 adult specific patho-gen-free(SPF)female SD rats were divided into 4 groups:sham surgery group,scoliosis model group,scoliosis model+P300 group,and scoliosis model+P300+Nrf2-IN-3 group.The scoliosis model was established by removing the upper limbs and tail of the rats through surgery.In the scolio-sis model+P300 group,P300 was intravenously injected[15 mg/(kg·d),30 d]after model-ing.In the scoliosis model+P300+Nrf2-IN-3 group,P300[15 mg/(kg·d),30 d]and the Nrf2 inhibitor Nrf2-IN-3[23.5 mg/(kg·d),30 d]were intravenously injected after model-ing.After 30 days,the nucleus pulposus tissues from the T12-L1 intervertebral discs were collected,and the expression levels of SOX9,COL-Ⅱ,MMP-13,and ADAMTS-5 were determined using Western blotting.Results Compared with the control group,the IL-1 β group showed decreased cell viability,increased cell apoptosis,upregulated expression levels of TNF-α,IL-6,MMP-13,ADAMTS-5,p-P65,and p-IKBα,and downregulated expression levels of SOX9,COL-Ⅱ,IκBα,Nrf2,and HO-1(all P＜0.05).Compared with the IL-1 β group,the IL-1 β+P300 group showed increased cell viability,decreased cell apoptosis,decreased expression levels of TNF-α,IL-6,MMP-13,ADAMTS-5,p-P65,and p-IKBα,and increased expression levels of SOX9,COL-Ⅱ,IKBα,Nrf2,and HO-1(all P＜0.05).Compared with the sham surgery group,the scoliosis model group showed decreased expression levels of SOX9 and COL-Ⅱ,and increased ex-pression levels of MMP-13 and ADAMTS-5(all P＜0.05).Compared with the scoliosis model group,the scoliosis model+P300 group showed increased expression levels of SOX9 and COL-Ⅱ,and decreased expression levels of MMP-13 and ADAMTS-5(all P＜0.05).Compared with the scoliosis model+P300 group,the scoliosis model+P300+Nrf2-IN-3 group showed decreased ex-pression levels of SOX9 and COL-Ⅱ,and increased expression levels of MMP-13 and ADAMTS-5(all P＜0.05).Conclusion P300 inhibits intervertebral disc degeneration by regulating the Nrf2/HO-1/NF-κB signaling pathway in the rat scoliosis model.
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