目的：探讨白藜芦醇对细菌脂多糖（LPS）耐受的人原单核细胞THP-1细胞中TNF-α启动子区的影响。方法：首先建立LPS耐受THP-1细胞模型后，然后用LPS分别刺激正常THP-1细胞（对照组）、LPS耐受THP-1细胞（耐受组）、白藜芦醇处理的LPS耐受THP-1细胞（耐受+白藜芦醇组），检测各组细胞TNF-α mRNA的表达及TNF-α启动子区各转录因子的结合情况。结果：LPS刺激后，TNF-α mRNA表达在对照组细胞迅速升高，耐受组缓慢升高，而耐受+白藜芦醇组略微升高，表达量明显低于前两组（均P<0.05）。染色体免疫沉淀（ChIP）分析显示，LPS刺激前，耐受组与耐受+白藜芦醇组p65及乙酰化p65（ace-p65）、RelB、G9a对TNF-p启动子区的结合量均明显高于对照组（均P<0.05），而p50的结合量各组细胞间无统计学差异（P>0.05）；LPS刺激后，p65和ace-p65对TNF-α启动子区的结合量在对照组明显增加，而在耐受+白藜芦醇组明显降低，G9a结合量在对照组量明显降低（均P<0.05），其余转录因子较刺激前无明显改变（均P>0.05）。结论：白藜芦醇可以抑制LPS耐受的THP-1细胞中TNF-α mRNA的表达，可能部分通过抑制p65/ace-p65对TNF-α启动子区的结合，因此可考虑将白藜芦醇用于治疗脓毒症的辅助治疗。
Effect of resveratrol on TNF-α promoter region in THP-1 cells tolerant to bacterial lipopolysaccharide
Objective: To investigate the effect of resveratrol on TNF-α promoter region in human monocyte THP-1 cells tolerated to bacterial lipopolysaccharide (LPS). Methods: The LPS-tolerated THP-1 cells were induced, next the untreated THP-1 cells (control group), LPS-tolerated THP-1 cells (tolerance group) and resveratrol treated LPS-tolerated THP-1 cells (tolerance plus resveratrol group) were stimulated with LPS respectively, and then, the TNF-α mRNA expression and the bindings of transcription factors to the promoter region of TNF-α were determined. Results: After LPS stimulation, the TNF-α mRNA level was increased rapidly in control group, and increased slowly in tolerance group, but were slightly decreased in tolerance plus resveratrol group, which was significantly lower than that in either former group (both P<0.05). Results of chromatin immunoprecipitation (ChIP) showed that before LPS simulation, the levels of p65 along with acetylated p65 (ace-p65), RelB and G9a binding to the promoter region of TNF-α in both tolerance group and tolerance plus resveratrol group were significantly higher than those in control group (all P<0.05), while the binding level of p50 had no significant difference among the three groups (P>0.05); after LPS simulation, the levels of p65 and ace-p65 binding to the promoter region of TNF-α were significantly increased in control group, but significantly decreased in tolerance plus resveratrol group, and level of G9a binding to the promoter region of TNF-α was decreased in control group (all P<0.05), and all other factors had no significant change compared with those before LPS stimulation (all P>0.05). Conclusion: Resveratrol can inhibit TNF-α mRNA expression in LPS-tolered THP-1 cells, which may be partially associated with its inhibiting the bindings of p65/ace-p65 to TNF-α promoter. So, Resveratrol may potentially be used in supplementary treatment of sepsis.