宿主细胞的脂筏在问号钩端螺旋体的进入和存活中起关键作用  

Host cell lipid raft plays a key role in L.interrogans entry and survival

作　　者：吕天宝 王佳琪 谢旭峰 宋宁 关钰颖 吴殿君 张文龙 曹永国 LYU Tianbao;WANG Jiaqi;XIE Xufeng;SONG Ning;GUAN Yuying;WU Dianjun;ZHANG Wenlong;CAO Yongguo(College of Veterinary Medicine,Jilin University,Changchun 130062,China)

机构地区：[1]吉林大学动物医学学院,吉林长春130062

出　　处：《中国兽医学报》2021年第5期962-967,共6页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31802261);中国博士后科学基金资助项目(2020M670860,BX20190141)。

摘　　要：钩端螺旋体病(钩体病)是由致病性钩端螺旋体(钩体)感染引起的一种全球广泛传播的自然疫源性人兽共患传染病。被钩体感染的人或动物可能会发展为严重的全身性疾病,严重者会导致死亡,但目前钩体进入细胞的机制尚不清楚。本试验探究了钩体(56601)进入细胞是否与脂筏相关。结果表明,J774A.1细胞和Vero细胞的脂筏被破坏后,其细胞内的钩体量明显降低,表明钩体进入细胞的过程与脂筏相关,并且钩体进入细胞依赖脂筏而与巨噬细胞的吞噬作用无关。透射电子显微镜、qPCR和Western blot结果显示,脂筏被破坏后加速了细胞内钩体的清除并减少了胞内钩体量,并且激活了细胞自噬。为了探究细胞清除钩体能力的增强是否与细胞自噬有关,采用雷帕霉素和甲基腺嘌呤(3-methyladenine, 3MA)分别刺激细胞,钩体培养计数结果显示,雷帕霉素处理组胞内钩体量明显降低,表明细胞自噬水平的提高有助于杀灭钩体。综上所述,钩体进入细胞需要脂筏,脂筏被破坏后,进入细胞的钩体量明显减少,并且细胞清除钩体的能力明显增高,这可能与细胞自噬水平的升高有关。Leptospirosis, caused by pathogenic Leptospira,is a widespread zoonotic disease.Human and animals infected with leptospires may develop severe systemic diseases, which can lead to death in severe cases, but the mechanism of leptospires entering macrophages is unclear.Our experiment explored whether the entry of pathogenic Leptospira(56601) into cells is related to lipid rafts.The results showed that the burden of leptospires in J774 A.1 cells and Vero cells significantly reduced after lipid rafts were disrupted, indicating that the process of leptospires entering cells was related to lipid rafts, moreover, the dependence of leptospires entering cells on lipid rafts was not related to the phagocytosis of macrophages.Transmission electron microscopy, qPCR and Western blot results showed that the disruption of lipid rafts accelerated the clearance of intracellular leptospires, reduced the burden of leptospires, and activated autophagy.In order to explore whether the enhancement of the ability of cells to remove leptospires is related to autophagy, the cells were treated with rapamycin and 3 MA respectively.The qPCR results showed that the burden of leptospires in the rapamycin treatment group was significantly reduced, indicating that the increase of autophagy level helps to kill leptospires.In summary, Leptospira requires lipid rafts to enter cells.After lipid rafts are disrupted, the burden of leptospires entering cells is significantly reduced, and the ability of cells to kill leptospires is significantly enhanced, which may be related to the improvement of autophagy.

关 键 词：钩端螺旋体 脂筏 细胞自噬 

分 类 号：S852.61[农业科学—基础兽医学] R535[农业科学—兽医学]