AMPK通过抑制STING调节干扰素免疫应答通路的研究

AMP-activated kinase inhibits IFN pathway through suppression of STING signaling

目的探索腺苷酸活化蛋白激酶(AMPK)与cGAS-STING通路之间的联系及其在先天免疫中扮演的角色。方法利用CRISPR/Cas9技术、蛋白质印迹、RT-qPCR等方法,探究AMPK对DNA相关免疫通路的调控机制。结果在HT-DNA和cGAMP刺激下,AMPK-/-细胞株的IFN-β的表达量明显高于野生型细胞株,但这种变化在RNA信号通路中并不明显;激活AMPK可以抑制细胞内的DNA信号通路;在DNA信号通路中,AMPK-/-细胞株相较于野生型细胞株,STING在RNA和蛋白水平上都明显升高,即AMPK对cGAS-STING通路的抑制很可能是通过抑制STING起作用。结论AMPK在调节cGAS-STING介导的干扰素免疫应答中起重要作用。

Objective AMP-activated protein kinase(AMPK)is a major regulator of cellular energy metabolism and plays a very key role in cell proliferation,differentiation and apoptosis.Immune responses to invasion of foreign pathogenic microorganisms is a very energy-intensive process.However,the link between AMPK and the cGAS-STING pathway,and its role in innate immunity remains unclear.Methods We used CRISPR/Cas9 technology,Western blot and RT-qPCR to explore the AMPK regulatory role involved in DNA-related innate immune pathway.Results In the stimulation of HT-DNA and cGAMP,the expression level of IFN-β in the AMPK^-/- cell lines was significantly higher than that of the wild type cell line,but this change was not obvious in the RNA signaling pathway.Activation of AMPK can also inhibit DNA signaling pathway.Compared with wild type cell lines,AMPK^-/- cell lines showed the increases in STING expression at both RNA and protein levels,suggesting that AMPK inhibited the cGAS-STING pathway by inhibiting STING.Conclusion These findings provide new insights into the mechanism of AMPK function in the innate immune reponse.