首页|绿原酸抑制LPS诱导的RAW264.7细胞炎症及机制探索

绿原酸抑制LPS诱导的RAW264.7细胞炎症及机制探索

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目的 明确绿原酸对脂多糖(lipopolysaccharide,LPS)诱导的巨噬细胞RAW264.7炎症的抗炎作用,揭示其可能的分子机制.方法 以CCK-8法检测不同浓度绿原酸干预后RAW264.7的细胞活性;采用LPS刺激巨噬细胞RAW264.7,预制细胞炎性状态,设置空白对照组(K组,n=3)、模型对照组(L组,n=3)和实验组(S组,n=3)分别给药,动态观察细胞形态;分别于24h及48h获取细胞沉淀及上清,以酶联免疫吸附(enzyme-linked immunosorbent assay,ELISA)法检测细胞上清中炎性细胞因子白细胞介素-1 β(interleukin-1 beta,IL-1β)、单核细胞趋化蛋白-1(monocyte chemoattractant protein-1,MCP-1)及精氨酸酶-1(arginase-1,Arg-1)的浓度;采用实时荧光定量 PCR(quantitative real-time polymerase chain reaction,RT-qPCR)法检测前列腺素内过氧化物合酶 2(prostaglandin-endoperoxide synthase 2,PTGS2)、转化生长因子 β1(transforming growth factor-beta1,TGF-β1)、高迁移率族蛋白 1(high mobility group box-1,HMGB1)及核转录因子-κB(nuclear transcription factor-kappaB,NF-κB)的mRNA表达.结果 1 μg/ml的LPS成功预制RAW264.7细胞炎性状态;12.5~200.0μg/ml的绿原酸对RAW264.7无明显毒性,50μg/ml及200μg/ml的绿原酸对RAW264.7细胞有明显的增殖作用(P<0.05);与模型对照组比较,实验组上清中IL-1β蛋白含量明显降低,Arg-1的含量明显升高(P<0.05);与模型对照组比较,50μg/ml的绿原酸显著降低LPS诱导的炎性细胞中NF-κB、TGF-β1、PTGS2及HMGB1的mRNA表达(P<0.05).结论 绿原酸对LPS诱导的RAW264.7炎性反应有明确抑制作用,可能与调控HMGB1介导的NF-κB相关通路分子表达有关.
Chlorogenic Acid Inhibits LPS-induced RAW264.7 Cell Inflammation and its Mechanism
Objective To clarify the anti-inflammatory effect of chlorogenic acid on lipopolysaccharide(LPS)-induced inflamma-tion in macrophage RAW264.7,and to reveal its possible molecular mechanism.Methods The cellular activity of RAW264.7 after the intervention of different concentrations of chlorogenic acid was detected by CCK-8 method;the macrophage RAW264.7 was stimulated by LPS to preset cellular inflammatory state.The blank control group(K group,n=3),the model control group(L group,n=3),and the experimental group(S group,n=3)were administered separately,and the cell morphology was dynamically observed;cell precipita-tion and supernatants were obtained at 24 h and 48h,respectively.The concentrations of inflammatory cytokine interleukin(IL)-1β,monocyte chemoattractant protein-1(MCP-1),and arginase-1(Arg-1)in the cell supernatants were detected by enzyme-linked immunosorbent assay(ELISA);the mRNA expression of prostaglandin endoperoxide synthase 2(PTGS2),transforming growth factor-β1(TGF-β1),high mobility histone 1(HMGB1),and nuclear transcription factor-κB(NF-κB)were detected by quantitative real-time PCR(RT-qPCR).Results RAW264.7 cellular inflammatory state was successfully preset with 1 μg/ml LPS;12.5~200.0μg/ml chlorogenic acid had no distinct toxicity on RAW264.7.Chlorogenic acid at 50μg/ml and 200μg/ml had obvious proliferation effects on RAW264.7 cells(P<0.05).Compared with the model control group,the content of IL-1 β protein in the supernatant of the experimental group was significantly decreased,while the content of Arg-1 was significantly increased(P<0.05).Compared with the model control group,50μg/ml of chlorogenic acid significantly decreased the mRNA expressions of NF-κB,TGF-β1,PTGS2,and HMGB1 in LPS-induced inflammatory cells(P<0.05).Conclusion Chlorogenic acid has a distinct inhibitory effect on LPS-induced RAW264.7 inflammatory response,which may be achieved by regulating molecular expression of the HMGB1-mediated NF-κB pathway.

Chlorogenic acidLipopolysaccharideRAW264.7Anti-inflammatory

周云鑫、刘柯、丁军颖

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100010 首都医科大学附属北京中医医院、北京市中医药研究所、中医感染性疾病基础研究北京市重点实验室

绿原酸 脂多糖 RAW264.7 抗炎

北京市自然科学基金北京市高层次公共卫生技术人才建设项目

7182071学科骨干-01-009

2024

医学研究杂志
中国医学科学院

医学研究杂志

CSTPCD
影响因子:0.702
ISSN:1673-548X
年,卷(期):2024.53(1)
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