Effects of caspase-11 non-canonical inflammasome on Leptospira interrogans-induced secretion of inflammatory cytokines in J774A.1 cells

Q4 Immunology and Microbiology
Xiaojuan Wang, Ziyu Xiao, Y. Liu, M. Wang, Junfei Huang, Q. Ma, Yue Wang, Xu Chen, H. Yong, F. Hong, Shijun Li
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引用次数: 0

Abstract

Objective To investigate the effects of caspase-11 non-canonical inflammasome on the Leptospira interrogans (L.interrogans)-induced secretion of inflammatory cytokines in J774A.1 cells. Methods Murine mononuclear macrophage cells (J774A.1) were infected with L. interrogans strain 56601. Expression of caspase-11, IL-1β, IL-1α and IL-18 at mRNA level in J774A.1 cells were detected by real-time RT-PCR. The levels of caspase-11, IL-1β, IL-1α and IL-18 in the culture supernatants of J774A.1 cells were detected by ELISA. Results Real-time RT-PCR showed that caspase-11 expression at mRNA level was 5.12, 14.21, 8.94, 14.06, 18.58 and 0.93 times of that in uninfected cells after 1, 2, 4, 8, 12 and 24 h of L. interrogans infection, and respectively decreased to 0.10, 0.07, 0.10, 0.09, 0.07 and 0.45 times after caspase-11 inhibitor intervention (P<0.05). Expression of IL-1β, IL-1α and IL-18 at mRNA level was significantly increased after infection (P<0.05). After the intervention with caspase-11 inhibitor, IL-1β mRNA decreased to 0.05, 0.03, 0.02, 0.05, 0.06 and 0.02 times (P<0.05); IL-1α mRNA decreased to 0.14, 0.07, 0.15, 0.10, 0.03 and 0.06 times (P<0.05); IL-18 mRNA decreased to 0.08, 0.10, 0.16, 0.18, 0.10 and 0.07 times (P<0.05). ELISA results showed that the expression of caspase-11, IL-1β, IL-1α and IL-18 at protein level was significantly increased. After the intervention with caspase-11 inhibitor, caspase-11 level decreased to 43.07, 41.64, 51.96, 86.56, 105.36, and 129.95 pg/ml (P<0.05); IL-1β level decreased to 15.01, 14.19, 68.02, 31.20, 173.13 and 104.98 pg/ml (P<0.05); IL-1α level decreased to 12.14, 15.40, 38.01, 21.97, 24.48 and 27.09 pg/ml (P<0.05); IL-18 level decreased to 96.27, 102.21, 85.34, 116.28, 155.36 and 114.03 pg/ml (P<0.05). Conclusions Caspase-11 non-canonical inflammasome was involved in the mediation of IL-1β, IL-1α and IL-18 secretion in mouse mononuclear macrophages after L. interrogans infection. Key words: Leptospira interrogans; Caspase-11 non-canonical inflammasome; Inflammatory cytokine; Macrophage; Gene silencing
caspase-11非规范炎性小体对钩端螺旋体诱导的J774A炎性细胞因子分泌的影响。1细胞
目的探讨胱天蛋白酶-11非典型炎症小体对钩端螺旋体诱导J774A1细胞分泌炎性细胞因子的影响。方法用小白鼠J774A1单核巨噬细胞感染询问乳杆菌56601株。实时RT-PCR检测J774A1细胞中半胱氨酸天冬氨酸蛋白酶11、IL-1β、IL-1α和IL-18的mRNA表达。用ELISA法检测J774A1细胞培养上清液中胱天蛋白酶11、白细胞介素1β、白细胞因子1α和白细胞介因子18的水平。结果实时RT-PCR结果显示,在询问乳杆菌感染1、2、4、8、12和24小时后,胱天蛋白酶-11的mRNA表达水平分别是未感染细胞的5.12、14.21、8.94、14.06、18.58和0.93倍,在胱天蛋白酶11抑制剂干预后分别降至0.10、0.07、0.10、0.09、0.07和0.45倍(P<0.05),感染后IL-1α和IL-18 mRNA水平显著升高(P<0.05),经胱天蛋白酶11抑制剂干预后,IL-1βmRNA水平分别下降至0.05、0.03、0.02、0.05、0.06和0.02倍(P<0.01);IL-1αmRNA分别下降到0.14、0.07、0.15、0.10、0.03和0.06倍(P<0.05);IL-18mRNA分别下降到0.08、0.10、0.16、0.18、0.10和0.07倍(P<0.05)。ELISA结果显示,胱天蛋白酶11、IL-1β、IL-1α和IL-18在蛋白水平上的表达显著增加。胱天蛋白酶-11抑制剂干预后,胱天蛋白酶11水平分别降至43.07、41.64、51.96、86.56、105.36和129.95 pg/ml(P<0.05);IL-1β水平分别降至15.01、14.19、68.02、31.20、173.13和104.98 pg/ml(P<0.05);IL-1α水平分别降至12.14、15.40、38.01、21.97、24.48和27.09 pg/ml(P<0.05);白细胞介素-18水平分别降至96.27、102.21、85.34、116.28、155.36和114.03pg/ml(P<0.05)。关键词:钩端螺旋体;Caspase-11非典型炎症小体;炎症细胞因子;巨噬细胞;基因沉默
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来源期刊
中华微生物学和免疫学杂志
中华微生物学和免疫学杂志 Immunology and Microbiology-Virology
CiteScore
0.50
自引率
0.00%
发文量
6906
期刊介绍: Chinese Journal of Microbiology and Immunology established in 1981. It is one of the series of journal sponsored by Chinese Medical Association. The aim of this journal is to spread and exchange the scientific achievements and practical experience in order to promote the development of medical microbiology and immunology. Its main contents comprise academic thesis, brief reports, reviews, summaries, news of meetings, book reviews and trends of home and abroad in this field. The distinguishing feature of the journal is to give the priority to the reports on the research of basic theory, and take account of the reports on clinical and practical skills.
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