OmpR对伤寒沙门菌巨噬细胞内生存能力的影响

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摘要目的: 研究OmpR对伤寒沙门菌巨噬细胞内生存能力的影响及相关分子机制。方法: 利用空载体对照株（WT-pBAD33）、ompR缺陷株（ΔompRpBAD33）和 ompR回补株（C-ΔompR）进行巨噬细胞THP1胞内生存能力实验，探究伤寒沙门菌OmpR对于巨噬细胞内生存能力的影响。进一步用实时荧光定量PCR（qRTPCR）、β半乳糖苷酶活性、凝胶阻滞实验（EMSA）分析伤寒沙门菌OmpR对ssrA、ssrB和spiC的调控作用。结果： ΔompRpBAD33巨噬细胞内生存能力显著低于WTpBAD33，C-ΔompR部分恢复了其生存能力；qRT-PCR和β-半乳糖苷酶活性实验结果表明，ΔompR-pBAD33中巨噬细胞内生存相关基因ssrA、ssrB和spiC的转录水平较WT-pBAD33明显下降; 体外表达纯化His-OmpR蛋白后进行的EMSA表明，OmpR可直接与ssrA、ssrB和spiC基因启动子区域结合。结论： OmpR通过直接调节巨噬细胞内生存相关基因ssrA、ssrB和spiC的表达，从而增强伤寒沙门菌巨噬细胞内生存能力。

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	魏丹丹
	谢中艺
	杨帆帆
	林立萍
	黄新祥
	张盈

关键词 ：伤寒沙门菌, OmpR, 巨噬细胞内生存能力

Abstract：Objective: To study the effect of OmpR on the viability of Salmonella enterica serovar Typhi (S. Typhi) in macrophages and its related molecular mechanisms. Methods: To investigate the effect of OmpR on the viability of S. Typhi in macrophages, the empty vector strain (WT-pBAD33), the ompR deletion strain (ΔompRpBAD33) and the ompR complementary strain (C-ΔompR) were used to carry out the THP1 intracellular viability assay. In addition, qRTPCR, βgalactosidase activity and EMSA were used to analyze the rol- of OmpR in the regulation of ssrA, ssrB and spiC expression. Results: The viability of ΔompRpBAD33 in macrophages was significantly lower than that of WTpBAD33, and C-ΔompR partially restored its viability. The results of qRT-PCR and βgalactosidase activity showed that the transcript levels of ssrA, ssrB and spiC in ΔompR-pBAD33 were significantly lower than those in WT-pBAD33. The EMSA showed that the purified HisOmpR bound in vitro directly to the promoter regions of ssrA, ssrB and spiC. Conclusion: OmpR enhances the viability of S. Typhi in macrophages by directly regulating the expression of ssrA, ssrB and spiC.

收稿日期: 2019-11-12

基金资助:中国博士后科学基金资助项目（2018M642186）

作者简介: 魏丹丹（1990—），女，硕士研究生

引用本文:

魏丹丹，谢中艺，杨帆帆，林立萍，黄新祥，张盈. OmpR对伤寒沙门菌巨噬细胞内生存能力的影响[J]. 江苏大学学报:医学版, 2020, 30(02): 149-153. WEI Dan-dan, XIE Zhong-yi, YANG Fan-fan, LIN Li-ping, HUANG Xin-xiang, ZHANG Ying.

Effect of OmpR on the viability of Salmonella enterica serovar Typhi in macrophages

. Journal of Jiangsu University(Medicine Edition), 2020, 30(02): 149-153.

链接本文:

http://zzs.ujs.edu.cn/xbyxb/CN/ 或 http://zzs.ujs.edu.cn/xbyxb/CN/Y2020/V30/I02/149

［1］Besser JM. Salmonella epidemiology: A whirlwind of change\[J\]. Food Microbiol, 2018, 71:55-59.

［2］Crump JA, Sj lundKarlsson M, Gordon MA, et al. Epidemiology, clinical presentation, laboratory diagnosis, antimicrobial resistance, and antimicrobial management of invasive Salmonella infections\[J\]. Clin Microbiol Rev, 2015, 28(4):901-937.

［3］Chakraborty S, Winardhi RS, Morgan LK, et al. Noncanonical activation of OmpR drives acid and osmotic stress responses in single bacterial cells\[J\]. Nat Commun, 2017, 8(1):1587.

［4］林立萍, 张连璐, 易周易, 等. OmpR对伤寒沙门菌侵袭上皮细胞能力的影响\[J\]. 江苏大学学报：医学版, 2019, 29(2):128-132.

［5］Zhao Z, Peng T, Oh JI, et al. A response regulator of the OmpR family is part of the regulatory network controlling the oxidative stress response of Rhodobacter sphaeroides\[J\]. Environ Microbiol Rep, 2019, 11(2):118-128.

［6］Tipton KA, Rather PN. An ompR/envZ twocomponent system ortholog regulates phase variation, osmotic tolerance, motility, and virulence in Acinetobacter baumannii strain AB5075\[J\]. J Bacteriol, 2017, 199(3):e00705-e00716.

［7］孙兰清, 左玲莉, 杨斯迪, 等. 沙门菌致病的奥秘\[J\]. 科学, 2019, 71(1):46-50.

［8］Pang X, Zhang R, Cheng G. Progress towards understanding the pathogenesis of dengue hemorrhagic fever\[J\]. Virol Sin, 2017, 32(1):16-22.

［9］殷俊磊, 李求春, 焦新安. 沙门菌致病岛2 Ⅲ型分泌系统研究进展\[J\]. 微生物学报, 2016, 56(4):561-569.

［10］Tsai CN, Coombes BK. The role of the host in driving phenotypic heterogeneity in Salmonella\[J\]. Trends Microbiol, 2019, 27(6):508-523.

［11］Shotland Y, Kr mer H, Groisman EA. The Salmonella SpiC protein targets the mammalian Hook3 protein function to alter cellular trafficking\[J\]. Mol Microbiol, 2003, 49(6):1565-1576.

［12］Chakraborty S, Mizusaki H, Kenney LJ. A FRETbased DNA biosensor tracks OmpRdependent acidification of Salmonella during macrophage infection\[J\]. PloS Biol, 2015, 13(4):e1002116.