雷公藤甲素对TLR7激动剂Resiquimod诱导RAW264.7巨噬细胞自噬的影响  

The effect of triptolide on RAW264.7 macrophage autophagy induced by TLR7 agonist resiquimod

作　　者：吴艮艮 时乐[1] 沈星星 梁涛[1] WU Gengen;SHI Le;SHEN Xingxing;LIANG Tao(College of Pharmacy,Nanjing University of Chinese Medicine,Nanjing 210046,China)

机构地区：[1]南京中医药大学药学院,210046

出　　处：《免疫学杂志》2018年第10期880-885,共6页Immunological Journal

摘　　要：目的考察TLR7激动剂Resiquimod(R848)能否诱导RAW264.7细胞自噬以及雷公藤甲素对R848诱导的自噬的影响。方法用0.01、0.1、1μg/ml的TLR7激动剂R848诱导RAW264.7细胞12、24、36 h,Western blot检测自噬蛋白LC3II/I、P62、Beclin1和Atg5表达情况;RAW264.7细胞采用0.1μg/ml R848处理24 h诱导自噬后,给予5 ng/ml、10 ng/ml雷公藤甲素12、24 h对自噬相关蛋白LC3II/I、P62及通路蛋白AKT检测。结果 RAW264.7细胞采用0.1μg/ml R848处理24 h后自噬相关蛋白LC3II/I、P62、Beclin1和Atg5表达显著升高;加入不同质量浓度的TP后,LC3II/I、P62显著表达减少,通路蛋白AKT表达升高。结论TLR7激动剂R848能够诱导RAW264.7细胞自噬,而TP能够抑制R848诱导的自噬,并且该作用可能与PI3K/AKT信号通路有关。This study was designed to explore whether TLR7 agonist resiquimod （R848） induces autophagy in RAW264.7 cells and the effect of triptolide （TP） on R848-induced RAW264.7 cell autophagy. Firstly, RAW264.7 cells were induced with R848 （0.01, 0.1, 1μg/ml） for 12, 24 or 36 h, then the expression of autophagic proteins LC3Ⅱ/Ⅰ, P62, Beclinl and Atg5 was detected by Western blotting. Secondly, RAW264.7 cells stimulated with 0.1 μg/ ml R848 for 24 h were treated with 5, 10 ng/ml TP for 12 h or 24 h, and autophagy-related proteins LC3Ⅱ/Ⅰ, P62 and pathway protein AKT were detected. Data showed that the expression levels of autophagy-related proteins LC3Ⅱ/Ⅰ, P62, Beclin and Atg5 were significantly increased in RAW264.7 cells treated with 0.1μg/ml R848 for 24 h. After adding different concentrations of TP, the expression of LC3Ⅱ/Ⅰ and P62 was decreased, while AKT expression was elevated. In conclusion, TLR7 agonist R848 is capable of inducing autophagy in RAW264.7 cells, whereas TP is able to inhibit R848-induced autophagy, and this effect may be related to the PI3K/AKT signaling pathway.

关 键 词：TLR7 系统性红斑狼疮 自噬 雷公藤甲素 

分 类 号：R965.1[医药卫生—药理学]