【摘要】：目的探討通過TSA抑制組蛋白去乙酰化水平,研究組蛋白去乙�；瘜�(duì)軟骨細(xì)胞表型相關(guān)基因的影響及其機(jī)制,從表觀遺傳學(xué)角度為維持軟骨細(xì)胞表型提供新的思路。方法體外培養(yǎng)人關(guān)節(jié)軟骨細(xì)胞,建立軟骨細(xì)胞去分化模型,采用不同濃度TSA刺激,在不同時(shí)間點(diǎn)收集細(xì)胞,提取總RNA,利用qRT-PCR檢測(cè)Wnt-5a、SOX-9、COL-Ⅱ、COL-Ⅰ的mRNA表達(dá)情況。利用免疫熒光及Westerblot進(jìn)行COL-Ⅱ蛋白表達(dá)水平的檢測(cè)。結(jié)果與對(duì)照組比較,TSA(0.25~1.0μmol/L)顯著降低COL-ⅡmRNA表達(dá)水平及蛋白表達(dá)水平,升高Wnt-5a和SOX-9 mRNA表達(dá)水平;抑制Wnt-5a信號(hào)通路減弱TSA對(duì)COL-Ⅱ的抑制效應(yīng)。結(jié)論 TSA通過激活Wnt-5a和SOX-9從而抑制COL-Ⅱ的表達(dá),導(dǎo)致軟骨細(xì)胞表型喪失。因此,組蛋白去乙酰化通過降低Wnt-5a和SOX-9,升高COL-Ⅱ的表達(dá)水平,發(fā)揮維持軟骨細(xì)胞表型的作用。
[Abstract]:Objective to investigate the effect of TSA inhibition histone deacetylation on the phenotypic related genes of cartilage cells and its mechanism, and to provide a new idea for maintaining the phenotype of cartilage cells from the perspective of epigenetics. Methods Human articular cartilage cells were cultured in vitro and the model of cartilage dedifferentiation was established. The cells were collected at different time points stimulated by different concentrations of TSA. The total RNA, was extracted and the mRNA expression of Wnt-5a,SOX-9,COL- 鈪，

本文編號(hào)：2522302