寨卡病毒(Zika virus，ZIKV)为蚊媒传播病原体，感染该病毒可引起寨卡病毒病，目前尚无特异治疗药物。自噬是细胞维持自身稳态、促进胞内物质能量再利用的过程，但常被一些病毒劫持以促进自身复制。本研究以人肝癌细胞系Huh7为靶细胞，利用mTagRFP-mWasabi-LC3双荧光报告系统检测自噬体的形成；蛋白免疫印迹法检测细胞感染ZIKV不同时间后自噬标记蛋白LC3和P62的表达情况；用自噬抑制剂(渥曼青霉素、氯喹)、Beclin-1特异的短发夹RNA(short hairpin RNA，shRNA)和自噬激动剂雷帕霉素预处理细胞，再采用蛋白免疫印迹法和病毒空斑法检测其对病毒蛋白表达和胞外子代病毒含量的影响；采用蛋白免疫印迹法检测参与ZIKV感染自噬调节的信号通路。结果显示：ZIKV感染Huh7细胞后，LC3阳性自噬体的数量明显增多；自12 h开始，LC3-II的表达水平显著升高，P62蛋白表达水平在感染48 h后显著降低；两种自噬抑制剂处理和干扰Beclin-1表达均能有效抑制ZIKV感染和子代病毒的产生，而雷帕霉素处理增加了胞外子代病毒含量(P<0.05)；ZIKV感染增强了腺苷酸活化蛋白激酶(adenosine 5’-monophosphate-activated protein kinase，AMPK)和TSC2发生磷酸化，而哺乳动物雷帕霉素靶标(mammalian target of rapamycin，mTOR)的磷酸化水平下调。结果表明，ZIKV感染通过激活Huh7细胞中的AMPK/TSC2/mTOR信号通路诱导自噬，并利用自噬促进病毒复制和子代病毒的产生，这为研制以自噬相关分子为靶点的抗寨卡病毒药物提供了理论和实验依据。

Zika virus (ZIKV) is a kind of mosquito-borne pathogen. Its infection can cause Zika virus disease. There is currently no specific treatment. Autophagy is a process in which cells maintain their own homeostasis and promote the reuse of intracellular substances and energy, but it is often hijacked by some viruses to promote their own replication. In this study, human hepatoma cell line (Huh7) was used as the target cell, and the mTagRFP-mWasabi-LC3 dual fluorescent reporter system was used to detect the formation of autophagosomes. Western blotting was used to detect the expression of autophagy markers LC3 and P62 proteins after ZIKV infection at different time points. Cells were pretreated with autophagy inhibitors (wortmannin and chloroquine), Beclin-1 specific short hairpin RNA (shRNA) and autophagy inducer rapamycin. Then their effects on viral protein expression and extracellular progeny virus titer were detected by Western blotting and virus plaque assay. The signaling pathways involved in the regulation of autophagy in ZIKV infection were detected by Western blotting. The results showed that the number of LC3-positive autophagosomes increased significantly in Huh7 cells after infection by ZIKV. The expression of LC3-II increased significantly from 12 h post infection, and P62 protein was significantly down-regulated at 48 h post infection. Both autophagy inhibitor treatments and interference of Beclin-1 expression could effectively inhibited ZIKV infection and the production of progeny viruses. Rapamycin treatment increased the content of extracellular progeny viruses (P<0.05). ZIKV infection enhanced the phosphorylation of adenosine 5’-monophosphate-activated protein kinase (AMPK) and TSC2, and reduced the phosphorylation of mammalian target of rapamycin (mTOR). The results demonstrate that ZIKV infection induces autophagy by activating the AMPK/TSC2/mTOR signaling pathway in Huh7 cells, and utilizes autophagy to promote virus replication and progeny virus production. It provides theoretical and experimental basis for the development of anti-ZIKV drugs based on autophagy-related molecules.