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目的:本研究以RAW264.7细胞系为研究对象,烧伤血清刺激建立烧伤细胞模型,在体外研究HSF1的抗炎症机制。方法:Westernblot法检测烧伤血清刺激前后RAW264.7细胞中HSF1与NF-κB表达,凝胶阻滞法(EMSA)检测HSF1与炎症因子HMGB1和抗炎症因子IL-10基因启动子区相互作用,同时检测NF-κB是否参与上述的相互作用。结果:正常培养条件下的RAW264.7细胞几乎不表达HSF1,少量表达NF-κB,加入烧伤血清刺激后大量表达HSF1和NF-κB。HSF1与HMGB1和IL-10基因的启动子区HSE有相互作用。NF-κB不参与HSF1与HMGB1和IL-10基因启动子区HSE的结合。结论:HSF1与HMGB1和IL-10的启动子区HSE结合,抑制HMGB1的表达,促进IL-10的表达。在炎症反应中HMGB1和IL-10的表达不直接受NF-κB的调节。
OBJECTIVE: In this study, RAW264.7 cell line was used as the research object, and burn serum was used to stimulate the establishment of a burn cell model to study the anti-inflammatory mechanism of HSF1 in vitro. Methods: Western blotting was used to detect the expression of HSF1 and NF-κB in RAW264.7 cells before and after stimulation with burn serum. The interaction between HSF1 and HMGB1 and anti-inflammatory IL-10 gene promoter region was detected by EMSA. It is tested whether NF-κB is involved in the above interaction. Results: Under normal culture conditions, RAW264.7 cells expressed almost no HSF1 and a small amount of expression of NF-κB. After burn serum was added, HSF1 and NF-κB were abundantly expressed. HSF1 interacts with HSE in the promoter region of HMGB1 and IL-10 genes. NF-κB is not involved in the binding of HSF1 to HSE in the HMGB1 and IL-10 gene promoter regions. Conclusion: HSF1 binds to HSE in the promoter region of HMGB1 and IL-10, inhibits the expression of HMGB1 and promotes the expression of IL-10. The expression of HMGB1 and IL-10 in the inflammatory response is not directly regulated by NF-κB.