Inhibition of chloroquine on TLR4-MyD88-independent signaling pathway in lipopolysaccharide/endotoxin-induced RAW264.7 cells

王艳艳; 刘鑫; 曹红卫; 王宁; 鲁永玲; 郑江

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Inhibition of chloroquine on TLR4-MyD88-independent signaling pathway in lipopolysaccharide/endotoxin-induced RAW264.7 cells

更新时间：2026-01-09

- Inhibition of chloroquine on TLR4-MyD88-independent signaling pathway in lipopolysaccharide/endotoxin-induced RAW264.7 cells
- Journal of Army Medical University Vol. 32, Issue 9, Pages: 869-873(2010)
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  CLC： R392.32;R631.202;R978.61
- Published：2010
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王艳艳,刘鑫,曹红卫,王宁,鲁永玲,郑江.氯喹对脂多糖/内毒素诱导的RAW264.7细胞TLR4-MyD88非依赖信号转导途径的抑制作用[J].陆军军医大学学报(原第三军医大学学报),2010,32(09):869-873.

　Wang Yanyan,Liu Xin,Cao Hongwei,Wang Ning,Lu Yongling,Zheng Jiang.Inhibition of chloroquine on TLR4-MyD88-independent signaling pathway in lipopolysaccharide/endotoxin-induced RAW264.7 cells[J].J Amry Med Univ (J Third Mil Med Univ),2010,32(09):869-873.
王艳艳,刘鑫,曹红卫,王宁,鲁永玲,郑江.氯喹对脂多糖/内毒素诱导的RAW264.7细胞TLR4-MyD88非依赖信号转导途径的抑制作用[J].陆军军医大学学报(原第三军医大学学报),2010,32(09):869-873. DOI：

　Wang Yanyan,Liu Xin,Cao Hongwei,Wang Ning,Lu Yongling,Zheng Jiang.Inhibition of chloroquine on TLR4-MyD88-independent signaling pathway in lipopolysaccharide/endotoxin-induced RAW264.7 cells[J].J Amry Med Univ (J Third Mil Med Univ),2010,32(09):869-873. DOI：

摘要

目的探讨氯喹对脂多糖/内毒素(lipopolysaccharide/endotoxin

LPS)诱导TLR4-MyD88非依赖途径的抑制作用。方法 MTT法检测不同浓度氯喹(5、10、20、40、60 μg/ml)对RAW264.7细胞活性的影响，ELISA检测培养体系中TNF-α、IL-6在2、6、12、18、24 h的释放情况，RT-PCR检测TNF-α、IL-6、TLR4、TRAM mRNA的表达，EMSA检测NF-κB的活性，Western blot检测IκBα磷酸化水平和IRF3的降解情况。结果氯喹浓度小于40 μg/ml时对RAW264.7细胞活性无影响（P0.05）；应用氯喹预处理后LPS诱导的RAW264.7细胞TNF-α、IL-6在蛋白水平和核酸水平均受到明显抑制（P0.05），TLR4和TRAM在核酸水平均受到显著抑制（P0.01）；EMSA结果显示NF-κB的活化受到抑制，而Western blot检测显示IκBα磷酸化及IRF3的降解也受到抑制。结论氯喹对LPS诱导的RAW264.7细胞TLR4-MyD88非依赖信号转导途径具有抑制作用。

Abstract

Objective To study the inhibitory effect of chloroquine on TLR4-MyD88-independent signaling pathway lipopolysaccharide/endotoxin (LPS)-induced RAW264.7 cells. Methods Effect of various chloroquine (5、10、20、40、60 μg/ml) concentrations on viability of RAW264.7 cells was detected by MTT test. TNF-α and IL-6 levels in culture supernatant were measured at 2

24 h by ELISA

and RT-PCR for the detection of expressions of TNF-α

IL-6，TLR4 mRNA and TRAM mRNA. EMSA was used to detect NF-κB activity. IκBα phosphorylation and IRF3 degradation were assayed by Western blotting. Results Chloroquine had no effect on the viability of RAW264.7 cells at a concentration of less than 40 μg/ml （P0.05）. The expression of TNF-α

IL-6

TLR4 and TRAM in LES-induced RAW264.7 cells at protein and mRNA level was significantly inhibited (P0.05

P0.01). EMSA showed that LPS inhibited the activity of NF-κB and Western blotting showed that LPS could suppress the phosporylation of IκBα and the degradation of IRF3. Conclusion Chloroquine inhibits TLR4-MyD88-independent signaling pathway in LPS-induced RAW264.7 cells.

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