曲古菌素A抗骨关节炎作用的实验研究

发布时间：2019-01-07 14:14

【摘要】：目的研究曲古菌素A (TSA)对大鼠骨关节炎的影响,并初步探讨其作用机制。方法在体外实验中,利用白细胞介素-1β (IL-1β)诱导大鼠软骨细胞建立骨关节炎模型,以不同浓度TSA处理细胞,利用荧光定量PCR (qRT-PCR)检测实验组与对照组中MMP-1,3,13及TIMP-1的基因表达情况；利用Western Blot技术检测MMP-1,3,13和TIMP-1的蛋白水平及组蛋白H3、H4的乙酰化水平。在体内实验中,通过膝关节腔注射单碘乙酸钠(MIA)建立大鼠关节炎模型,分析膝关节标本形态学、组织学改变并检测软骨标本中目标基因和蛋白的表达水平。结果在体外实验中,TSA从基因和蛋白水平均可明显抑制IL-1p诱导的软骨细胞中MMP-1,3,13的表达并促进TIMP的表达,同时升高组蛋白乙酰化水平。而单独用TSA处理仅促进TIMP-1的表达,对MMPs无明显影响。大体标本的形态学和组织学结果提示TSA组的软骨病变较OA组明显减轻,标本的基因及蛋白表达检测结果与体外实验结果一致。结论TSA对骨关节炎有潜在的治疗作用,其抗炎机制可能是通过提高乙酰化水平来促进TIMP-1的表达,从而抑制MMPs的表达及活性来发挥抗骨关节炎作用。
[Abstract]:Objective to study the effect of trabeculin A (TSA) on osteoarthritis in rats and to explore its mechanism. Methods in vitro, osteoarthritis model was induced by interleukin-1 尾 (IL-1 尾) in rat chondrocytes. The cells were treated with different concentrations of TSA, and MMP-1,3, in experimental group and control group was detected by fluorescence quantitative PCR (qRT-PCR). 13 and TIMP-1 gene expression; The protein levels of MMP-1,3,13 and TIMP-1 and the acetylation level of histone H _ 3 H _ 4 were detected by Western Blot. In vivo, a rat model of arthritis was established by injecting monoiodoacetate (MIA) into knee joint cavity. The morphology, histological changes and expression level of target gene and protein in articular cartilage were analyzed. Results in vitro, TSA could significantly inhibit the expression of MMP-1,3,13 and promote the expression of TIMP in chondrocytes induced by IL-1p from both gene and protein levels, and increase the level of histone acetylation. TSA alone only promoted the expression of TIMP-1, but had no significant effect on MMPs. The morphological and histological results showed that the cartilage lesion in TSA group was significantly less than that in OA group, and the results of gene and protein expression were consistent with those in vitro. Conclusion TSA has a potential therapeutic effect on osteoarthritis and its anti-inflammatory mechanism may be to increase the level of acetylation to promote the expression of TIMP-1 and thus inhibit the expression and activity of MMPs to play the role of anti-osteoarthritis.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R684.3

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