糖皮质激素激活自噬对软骨细胞衰老的影响

发布时间：2018-12-25 12:50

【摘要】：目的:探讨糖皮质激素对关节软骨细胞衰老和自噬影响,并分析自噬在其中的作用及机制。方法:培养SD大鼠软骨细胞,阿利辛染色和collagen II免疫组化进行鉴定。使用不同浓度的地塞米松作用于软骨细胞不同的时间后,Lyso Tracker Red和MDC染色观察自噬囊泡的生成,Western blot检测LC3、beclin-1、P62、p70S6K和4EBP1蛋白表达的变化。β-半乳糖苷酶染色分析糖皮质激素对软骨细胞衰老的影响,并使用自噬抑制剂来探讨自噬对地塞米松作用下软骨细胞衰老的作用。结果:地塞米松可以显著提高软骨细胞的自噬水平,尤其是在作用4 d后,LC3-II表达随着地塞米松浓度的增加而升高,而P62和beclin-1的表达也验证自噬水平的升高。Lyso Tracker Red和MDC染色发现软骨细胞中自噬泡的生成随着地塞米松浓度的升高而增加。m TOR信号通路上的p70S6K和4EBP1的磷酸化均被地塞米松抑制。更重要的是,地塞米松可以引起软骨细胞的衰老,而使用3-MA抑制自噬后,细胞进一步衰老。结论:糖皮质激素激活软骨细胞中的自噬,并同时诱导软骨细胞的衰老,而自噬在这过程中对细胞的衰老可能具有代偿性的保护作用。同时糖皮质激素抑制m TOR信号通路,可能与自噬的激活相关。
[Abstract]:Aim: to investigate the effect of glucocorticoid on the aging and autophagy of articular chondrocytes, and to analyze the role and mechanism of autophagy. Methods: chondrocytes of SD rats were cultured and identified by alisin staining and collagen II immunohistochemistry. , Lyso Tracker Red and MDC staining of chondrocytes treated with dexamethasone at different concentrations for the detection of LC3,beclin-1,P62, by, Western blot The effect of glucocorticoid on chondrocyte senescence was analyzed by 尾 -galactosidase staining, and the effect of autophagy on chondrocyte senescence induced by dexamethasone was studied with autophagy inhibitor. Results: dexamethasone significantly increased the level of autophagy of chondrocytes, especially after 4 days of exposure, the expression of LC3-II increased with the increase of dexamethasone concentration. The expression of P62 and beclin-1 also confirmed the increase of autophagy level. Lyso Tracker Red and MDC staining showed that the formation of autophagy in chondrocytes increased the phosphorylation of p70S6K and 4EBP1 in. M TOR signaling pathway with the increase of dexamethasone concentration. All were inhibited by dexamethasone. More importantly, dexamethasone can cause chondrocyte senescence, while 3-MA inhibits autophagy and further senescence. Conclusion: glucocorticoid activates autophagy in chondrocytes and induces the aging of chondrocytes. Autophagy may play a compensatory role in protecting chondrocytes from aging. Meanwhile, glucocorticoid inhibits m TOR signaling pathway, which may be related to autophagy activation.
【作者单位】：温州医科大学附属第二医院关节外科;温州医科大学附属第二医院手术室;
【基金】：温州市科技局项目(No.Y20140582)
【分类号】：R363

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