Vorinostat promoted cartilage repair by the regulation of histone deacetylase-2
Objective To explore vorinostat,a histone deacetylase(HDAC)inhibitor,applied to promote cartilage repair and the underlying mechanisms.Methods Firstly,an in vivo experiment was conducted on rats,with a control group and a degeneration group.After successful modeling,cartilage samples were taken,and the rat cartilage was sliced for immunohistochemical HDAC-2 staining.Subse-quently,cDNA and protein were extracted for gene and protein detection of HDAC-2,respectively.Then,in the in vitro experiment section,rat chondrocytes were isolated and cultured in vitro,and set as control group,interleukin-1 group,and interleukin-1+vorinostat group.The gene expression of HDAC-2,type Ⅱcollagen(Col2a1),and proteoglycan(Acan)was detected by real-time quantitative reverse transcriptase-polymerase chain reaction(RT-qPCR),acetylation and total H3 histone protein expression were detected by Western blotting,and HDAC-2 protein was labeled by immunofluorescence.After determining the effect of vorinotat,a rat degeneration model was ultimately used,and a degeneration group and a degeneration+vorinotat group were set up.Samples were taken 7 days after injection and stained with safranin O and Ali-sin blue.The t-test was done for comparison between two groups.Results The distribution of HDAC-2 immunohistochemistry protein in the cartilage of the degenerative group rats was higher than that of the con-trol group,and its gene transcription activity was significantly increased(0.96±0.05 vs.1.34±0.12,t=5.83,P＜0.01).In vitro experiments showed that interleukin-1 induced a significant increase in the gene expression of HDAC-2(0.87±0.11 vs.1.88±0.64,t=3.12,P＜0.05)with increased immuno-fluorescence intensity.After treatment with vorinostat,the proportion of H3 histone acetylation in chondro-cytes increased(0.41±0.17 in the control group vs.1.09±0.12 in the vorinostat group,t=5.63,P＜0.01).Interleukin-1 could inhibit the transcriptional activity of Col2a1 and Acan in chondrocytes(4.86± 0.64 vs.2.28±0.49,t=5.53,P＜0.01;2.72±0.75 vs.1.29±0.31,t=3.06,P＜0.05),while the addition of vorinostat restored the transcriptional activity of Col2a1 and Acan in chondrocytes(2.28± 0.49 vs.4.05±0.72,t=3.53,P＜0.05;1.29±0.31 vs.2.58±0.58,t=3.38,P＜0.05).Subse-quent in vivo experiments were conducted to set up the degenerative group and the degenerative+vorinostat group,and it was observed that the cartilage defects,flatness,and collagen fiber distribution in the degen-erative+vorinostat group were superior to those in the degenerative group.Conclusion Vorinotat inhibits HDAC-2 activity,regulates histone deacetylation,increases the expression of type Ⅱ collagen and proteo-glycans,and promotes bone inflammation injury repair.
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