香烟烟雾提取物下调小鼠C2C12成肌细胞组蛋白去乙酰化酶2表达并增强炎性细胞因子释放

发布时间：2018-05-02 10:15

本文选题：香烟烟雾提取物 + CC成肌细胞　；参考：《细胞与分子免疫学杂志》2015年03期

【摘要】：目的观察香烟烟雾暴露后,小鼠C2C12成肌细胞内炎症介质、组蛋白去乙酰化酶2(HDAC2)和核因子κB(NF-κB)的变化。方法培养小鼠C2C12成肌细胞,用香烟烟雾提取物(CSE)进行处理。采用MTT法观察CSE对细胞生长的影响,以确定作用于细胞的合适浓度。将培养6~7 d的C2C12细胞接种到6孔板,分为对照组和(6.25、12.50、25.0)mL/L CSE组,培养24h。ELISA测定各组细胞上清液白细胞介素8(IL-8)和肿瘤坏死因子α(TNF-α)的含量;实时定量PCR(qRT-PCR)检测各组细胞HDAC2 mRNA的表达;Western blot法检测各组细胞中HDAC2的蛋白相对表达量;免疫共沉淀技术检测细胞内HDAC2/NF-κB复合物。结果 MTT结果提示终浓度为50 mL/L的CSE抑制C2C12细胞的生长;CSE刺激24h后,C2C12细胞IL-8和TNF-α释放增加,HDAC2的mRNA及蛋白相对表达量减少,并在一定程度上具有浓度依赖性;免疫共沉淀技术证实存在HDAC2/NF-κB复合物。结论 CSE下调C2C12细胞HDAC2表达,引起炎症因子释放增加。
[Abstract]:Objective to observe the changes of inflammatory mediators, histone deacetylase 2 (HDAC2) and nuclear factor- 魏 B (NF- 魏 B) in mouse C2C12 myoblasts after cigarette smoke exposure. Methods Mouse C2C12 myoblasts were cultured and treated with cigarette smoke extract. The effect of CSE on cell growth was observed by MTT method to determine the appropriate concentration. C2C12 cells cultured for 6 days were inoculated into 6 well plates and divided into two groups: control group and CSE group. The contents of interleukin-8 (IL-8) and tumor necrosis factor- 伪 (TNF- 伪) in supernatant of each group were measured by 24h.ELISA. The expression of HDAC2 mRNA was detected by real-time quantitative PCR qRT-PCR. the relative expression of HDAC2 protein was detected by Western blot assay, and the HDAC2 / NF- 魏 B complex was detected by immunoprecipitation. Results MTT results showed that 50 mL/L CSE inhibited the release of IL-8 and TNF- 伪 from C2C12 cells for 24 h and increased the relative expression of mRNA and protein of HDAC2 in a concentration-dependent manner. The existence of HDAC _ 2 / NF- 魏 B complex was confirmed by immunoprecipitation technique. Conclusion CSE can down-regulate the expression of HDAC2 in C2C12 cells and increase the release of inflammatory factors.
【作者单位】：广西医科大学第一附属医院呼吸内科;
【基金】：国家自然科学基金(81260012)
【分类号】：R563.9

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