论文部分内容阅读
目的通过检测山姜素作用下鼠巨噬细胞(RAW246.7)炎症基因启动子部位结合组蛋白乙酰化修饰状态以及炎症因子表达,初步揭示山姜素调节炎症因子表达的表观遗传学作用机制。方法采用不同质量浓度(10~200μg/ml)山姜素培养RAW246.7,培养结束后ELISA法检测养基中IL-6以及肿瘤坏死因子a(TNF-α)质量浓度,染色质免疫共沉淀(Chip-q PCR)法检测上述基因启动子片段结合的去乙酰化/乙酰化组蛋白(H3K9以及H3K14)表达比值,Pearson相关分析比值与炎症因子质量浓度的关联情况。结果山姜素呈剂量依赖性抑制RAW246.7细胞IL-6以及TNF-α表达,同时明显上调RAW246.7胞内IL-6以及TNF-α启动子片段结合的去乙酰化/乙酰化H3K9比值,且上述比值与炎症因子的表达水平呈负相关(P<0.05);山姜素作用下炎症基因启动子片断结合的去乙酰化/乙酰化H3K14比值无明显变化。结论山姜素可能通过诱导炎症基因的启动子部位组蛋白H3K9去乙酰化以调节炎症因子的合成,提示表观遗传学修饰很可能是山姜素干预炎症性疾病的重要机制。
OBJECTIVE: To investigate the epigenetic mechanism of alpinein-induced inflammatory cytokines by detecting the acetylation status of histone acetylation and the expression of inflammatory cytokines in the promoter region of inflammatory gene of rat macrophage RAW264.7 (RAW246.7) . Methods RAW246.7 was cultured with different concentrations of alpinin (10 ~ 200μg / ml). The concentrations of IL-6 and TNF-α in culture medium were detected by ELISA after culture. Chromatin co-immunoprecipitation The expression of deacetylated / acetylated histones (H3K9 and H3K14), Pearson correlation analysis and the mass concentration of inflammatory cytokines were detected by Chip-q PCR. Results Angauricine inhibited the expression of IL-6 and TNF-α in RAW246.7 cells in a dose-dependent manner and significantly increased the ratio of deacetylated / acetylated H3K9 bound by IL-6 and TNF-α promoter in RAW264.7 cells (P <0.05). There was no significant difference in the ratio of deacetylated / acetylated H3K14 between the combination of proinflammatory gene promoter and fragments of inflammatory gene promoter. Conclusion Pyrogallol may be the deacetylation of inflammatory cytokines through the induction of histone H3K9 at the promoter of inflammatory genes, suggesting that epigenetic modification may be an important mechanism of alpinein in inflammatory diseases.