山姜素通过促进H3K9去乙酰化抑制鼠巨噬细胞IL-6分泌的分子机制

发布时间：2018-05-12 23:14

本文选题：山姜素 + IL-　；参考：《免疫学杂志》2017年06期

【摘要】：目的通过检测鼠巨噬细胞(RAW246.7)IL-6启动子部位H3K9乙酰化修饰状态以及转录因子P40和CREB含量,初步揭示H3K9去乙酰化在山姜素调节RAW246.7炎症因子表达过程中的分子机制。方法 RAW246.7按照不同干预分为对照组、山姜素组(终质量浓度分别为50、100、200μg/ml),山姜素+GW9662组,山姜素+HDAC1(组蛋白去乙酰化酶1)或Pgene(对照质粒)Si RNA组,培养完成后ELISA法检测培基中IL-6水平,Western blot检测胞核PPAR、P40和CREB表达水平,并采用染色质免疫共沉淀(Chip-q PCR)检测RAW246.7 IL-6启动子部位结合的去乙酰化H3K9、P40以及CREB的相对表达水平。结果与对照组相比,山姜素呈剂量依赖性促进RAW246.7核内PPAR表达并遏制IL-6合成,可促进去乙酰化H3K9表达,并下调核内以及IL-6启动子结合的转录因子水平;GW9662遏制山姜素诱导的核内PPAR表达,逆转山姜素诱导的去乙酰化H3K9表达,同时恢复启动子部位、核内P40和CREB含量以及IL-6合成;HDAC1干扰对山姜素诱导的PPAR表达以及核内P40和CREB合成下降无显著影响,但可遏制山姜素诱导的去乙酰化H3K9表达,从而恢复IL-6启动子结合P40和CREB含量以及IL-6合成,但表达水平低于GW9662组。结论 PPAR激动剂山姜素通过激活去乙酰化酶促进IL-6启动子部位H3K9去乙酰化,以上改变可能干扰转录因子P40和CREB结合于启动子部位,从而干预IL-6表达,而H3K9去乙酰化不影响P40和CREB合成。
[Abstract]:Objective to explore the molecular mechanism of H3K9 deacetylation in the regulation of the expression of RAW246.7 inflammatory factors by detecting the H3K9 acetylation modification state and the contents of transcription factors P40 and CREB in the IL-6 promoter of rat macrophage RAW246.7. Methods according to different interventions, RAW246.7 was divided into control group (final concentration of 50100200 渭 g / ml), GW9662 group, histone deacetylase 1 (histone deacetylase 1) group or control plasmid RNA group. ELISA assay was used to detect the level of IL-6 and the expression of PPARN P40 and CREB were detected by Western blot, and the relative expression levels of deacetylated H3K9P40 and CREB in RAW246.7 IL-6 promoter were detected by chromatin immunoprecipitation (Chip-q PCR). Results compared with the control group, curcumin enhanced the expression of PPAR in RAW246.7 nucleus and inhibited the synthesis of IL-6 in a dose-dependent manner, and promoted the expression of deacetylated H3K9 in a dose-dependent manner. GW9662 inhibited the expression of PPAR, reversed the expression of deacetylated H3K9 induced by curcumin, and restored the site of promoter. The content of P40 and CREB in nucleus and the interference of IL-6 synthesis with HDAC1 had no significant effect on the expression of PPAR induced by curcumin and the decrease of synthesis of P40 and CREB in nucleus, but it could inhibit the expression of deacetylated H3K9 induced by curcumin. Thus, the levels of P40 and CREB binding and IL-6 synthesis of IL-6 promoter were restored, but the expression level was lower than that of GW9662 group. Conclusion the PPAR agonist curcumin promotes H3K9 deacetylation in the IL-6 promoter by activating deacetylase. These changes may interfere with the binding of transcription factor P40 and CREB to the promoter site and thus interfere with the expression of IL-6. However, H3K9 deacetylation did not affect the synthesis of P40 and CREB.
【作者单位】：湖南医药学院临床医学院;
【基金】：湖南省自然科学基金(2010FJ3161)
【分类号】：R392

【相似文献】