Helicobacter pylori Tipα protein induces IL-1β and IL-18 production in THP-1-drived macrophages through activation of NLRP3 inflammasome
XIAO Ling-qiao1, WANG Chao2, XIE Cheng-yuan2, LIU Shuo1, CHEN Guo-dong1, LI Jie3, YU Min-jun1, ZHAO Lan-hua1, HAN Liang1, ZHANG Yan1
1.Institute of Pathogenic Biology, University of South China, Hengyang 421001, China;
2.Class 1 of Distinguished Physicians Grade 2015, University of South China, Hengyang 421001, China;
3.Department of Biochemistry, Yongzhou Vocational Technical College, Yongzhou 425100, China
Abstract：We preliminarily investigated the role of NLRP3 inflammasome in Helicobacter pylori (H. pylori) Tipα-induced pro-inflammatory cytokines secretion in PMA-differentiated human acute monocytic leukemia cell line THP-1. PMA-differentiated THP-1 cells were treated with pure recombinant Tipα protein. The secretion levels of TNF-α, IL-1β and IL-18 in supernatant of culture medium were detected by ELISA. Then we blocked MyD88/NF-κB and NLRP3/Caspase-1 pathways by PDTC or the general ROS scavenger, NAC, respectively, and determined the secretion levels of proinflammatory cytokines and the expression levels of NLRP3 and Caspase-1. The results showed that Tipα protein can significantly induced the secretion of TNF-α, IL-1β and IL-18 in THP-1 cells in a time and dose-dependent manner. Levels of TNF-α, IL-1β and IL-18 approached their peaks at 6 h post-treatment by 40 μg/mL of Tipα protein (P<0.05). Moreover, the blockade of NF-κB signaling pathway by PDTC can inhibit the secretion of proinflammatory cytokines and the expression of NLRP3 and Caspase-1. When THP-1 cells were pre-treated with ROS scavenger NAC, the Tipα-induced increased IL-1β and IL-18 secretion was obviously eliminated (P<0.05), while TNF-α level had no significant difference, and the expression levels of Caspase-1 and NLRP3 also have a significant decrease. Our results demonstrated that Tipα can promote THP-1-drived macrophages to secrete proinflammatory cytokines TNF-α, IL-1β and IL-18, and the NLRP3/Caspase-1 pathway may be involved in the Tipα protein-induced IL-1β and IL-18 secretion.
肖玲巧, 王超, 谢成元, 刘硕, 陈国栋, 李杰, 余敏君, 赵兰华, 韩亮, 张艳. 幽门螺杆菌Tipα蛋白激活NLRP3炎性小体诱导THP-1细胞分泌IL-1β和IL-18[J]. 中国人兽共患病学报, 2017, 33(12): 1071-1076.
XIAO Ling-qiao, WANG Chao, XIE Cheng-yuan, LIU Shuo, CHEN Guo-dong, LI Jie, YU Min-jun, ZHAO Lan-hua, HAN Liang, ZHANG Yan. Helicobacter pylori Tipα protein induces IL-1β and IL-18 production in THP-1-drived macrophages through activation of NLRP3 inflammasome. Chinese Journal of Zoonoses, 2017, 33(12): 1071-1076.
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2017, Vol. 33 
