An in vitro study on the effect of histone deacetylation on type Ⅱ collage expression
Chinese Journal of Clinical Anatomy ›› 2015, Vol. 33 ›› Issue (6) : 667-671.
An in vitro study on the effect of histone deacetylation on type Ⅱ collage expression
Objective In the present study, we inhibited histone deacetylation of the chondrocytes by the treatment of TSA, aiming at investigating the effects of histone deacetylation on chondrocyte phenotype and related gene expression and the mechanisms，aiming at providing a new strategy to maintain the phenotype of chondrocytes from the perspective of epigenetics. Methods Human articular chondrocytes were cultured in vitro and a model of chondrocyte dedifferentiation was established first. Different concentrations of TSA were used for the stimulation and cells were collected at different time points for the extraction of total RNA. Quantitative real-time polymerase chain reaction (qRT-PCR) was performed for the detection of the expression of Wnt-5a, collagen typeⅠ(TypeⅠcollagen, COL-Ⅰ), collagen typeⅡ(TypeⅡcollagen, COL-Ⅱ) and SOX-9. Immunofluorescence and western blot were used to detect the protein expression of COL-Ⅱ. Results Comparing with control group, a concentration of 0.25~1.0 μmol/L TSA was sufficient to block protein and mRNA levels of typeⅠcollagen and typeⅡcollagen expression in primary culture chondrocytes, while it could promote the expression of Wnt-5a and SOX-9. Conclusions Inhibition of COL-Ⅱ expression which leads to the dedifferentiation of chondrocytes and the phenotype change might be mediated by the up-regulation of Wnt-5a and SOX-9. Therefore, histone deacetylation may elevate the expression level of COL-Ⅱ through down-regulation of Wnt-5a and SOX-9, which might play an important role in maintaining chondrocyte phenotype in vitro.
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