肝实质细胞抑制细菌增殖作用及机制研究

马娓; 舒俊傑; 陈地友; 王丽; 杨霞; 高强国

doi:10.16016/j.2097-0927.202306111

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肝实质细胞抑制细菌增殖作用及机制研究

基础医学 | 更新时间：2026-01-09

- 肝实质细胞抑制细菌增殖作用及机制研究
- Inhibitory effect and underlying mechanism of hepatocytes on bacterial proliferation
- 陆军军医大学学报 2023年45卷第21期页码：2206-2213
- 作者机构：
  
  1.400038 重庆，陆军军医大学(第三军医大学)基础医学院细胞生物学教研室
  2.400042 重庆，陆军特色医学中心野战外科研究部战伤感染与特需药品研究室
  3.400042 重庆，陆军特色医学中心放射科
  4.400042 重庆，陆军特色医学中心野战外科研究部军事交通伤防治研究室
- 作者简介：
  
  [杨霞，E-mail: oceanyx@126.com YANG Xia, E-mail: oceanyx@126.com
  [高强国，E-mail: qiangguogao@163.com GAO Qiangguo, E-mail: qiangguogao@163.com
- 基金信息：
  
  国家自然科学基金面上项目(81772717);国家自然科学基金面上项目(82172134);作者贡献声明马娓和舒俊傑负责方法设计、实验试剂材料准备、实验操作、数据整理及分析、实验结果可视化、初稿撰写等；陈地友、王丽负责实验操作、数据整理、实验结果可视化等；高强国、杨霞负责实验构思与设计、结果的审核、实验监督管理、稿件审阅和修改等
- DOI：10.16016/j.2097-0927.202306111
  中图分类号： R322.47;R329.28;R378
- 收稿：2023-06-28，
  
  修回：2023-08-03，
  
  纸质出版：2023-11-15
- 稿件说明：
移动端阅览
马娓, 舒俊傑, 陈地友, 等. 肝实质细胞抑制细菌增殖作用及机制研究[J]. 陆军军医大学学报, 2023,45(21):2206-2213.

Wei MA, Junjie SHU, Diyou CHEN, et al. Inhibitory effect and underlying mechanism of hepatocytes on bacterial proliferation[J]. Journal of Army Medical University, 2023, 45(21): 2206-2213.
马娓, 舒俊傑, 陈地友, 等. 肝实质细胞抑制细菌增殖作用及机制研究[J]. 陆军军医大学学报, 2023,45(21):2206-2213. DOI： 10.16016/j.2097-0927.202306111.

Wei MA, Junjie SHU, Diyou CHEN, et al. Inhibitory effect and underlying mechanism of hepatocytes on bacterial proliferation[J]. Journal of Army Medical University, 2023, 45(21): 2206-2213. DOI： 10.16016/j.2097-0927.202306111.

摘要

目的

探讨肝细胞通过直接吞噬和抗菌蛋白介导抑制细菌增殖的作用及其机制。

方法

利用正常小鼠FL83B肝细胞系建立大肠杆菌感染模型，以小鼠Kupffer细胞系为对照，观察肝细胞对大肠杆菌的增殖影响；建立细菌-细胞和(或)细胞上清共培养体系，通过Kupffer/FL83B细胞及其6、12、24 h的上清与大肠杆菌的共培养，检测大肠杆菌的致死情况，明确肝细胞直接抑菌效应；通过吞噬功能检测肝细胞的吞噬能力，qRT-PCR、Western blot检测相关基因的表达。

结果

将大肠杆菌与细胞共孵育，发现FL83B细胞与Kupffer细胞类似，可直接抑制大肠杆菌的增殖(

＜0.05)，且肝细胞自身保持高存活率；通过吞噬功能检测，明确了FL83B细胞可直接吞噬细菌颗粒；与对照组相比，感染后肝细胞内吞噬相关基因Fcgr1、Marco、Cdc42、RhoB mRNA上调，抗原递呈相关基因β2M mRNA上调，NF-κB、MAPK、STAT3通路活化，且介导抗菌作用的相关蛋白iNOS、NOX2、RhoB表达上调(

＜0.05)。

结论

小鼠FL83B肝细胞可直接通过吞噬作用和抗菌相关蛋白介导抑制细菌增殖。

Abstract

Objective

To explore the role and mechanism of hepatocytes in inhibiting bacterial proliferation by phagocytizing bacteria directly and mediating the expression of antibacterial proteins indirectly.

Methods

Cellular model of

Escherichia coli

(

E.coli

) infection was established in normal murine hepatocyte line FL83B and mouse Kupffer cells to determine the effects of the cells on the proliferation of bacteria. After the bacteria were co-cultured with Kupffer/FL83B cells and the 6-

12-

and 24-hour supernatants

respectively

the lethality of

E. coli

was determined for t

he direct bacteriostatic effect of the hepatocytes. The phagocytizing ability of FL83B cells were measured with phagocytosis reagent kit. qRT-PCR and Western blotting were employed to detect the expression of related molecules at mRNA and protein levels.

Results

When the hepatocyte were incubated with

E. coli

FL83B cells

as Kupffer cells

could directly inhibit the proliferation of the bacteria (

0.05) and maintained a high survival rate at same time. Phagocytic function test showed that FL83B cells could directly phagocytize bacterial particles. Compared with the FL83B cells without

E. coli

infection

the mRNA levels of phagocytosis-related genes Fcgr1

Marco

Cdc42

and RhoB were increased

that of antigen presentation related gene β2M was also upregulated

the NF-κB

MAPK

and STAT3 signaling pathways were activated

and the protein levels of antibacterial-related proteins iNOS

NOX2

and RhoB were also enhanced in the cells after infection (all

0.05).

Conclusion

Mouse FL83B hepatocytes possess the ability to inhibit bacteria through direct phagocytosis and indirect upregulation of antibacterial-related proteins.

关键词

Keywords

references

SON G, KREMER M, HINES I N. Contribution of gut bacteria to liver pathobiology[J]. Gastroenterol Res Pract, 2010, 2010: 453563. DOI: 10.1155/2010/453563.

BERG R D. Bacterial translocation from the gastrointestinal tract[J]. Trends Microbiol, 1995, 3(4): 149-154. DOI: 10.1016/S0966-842X(00)88906-4.

VAN DER HEIDE D, WEISKIRCHEN R, BANSAL R. Therapeutic targeting of hepatic macrophages for the treatment of liver diseases[J]. Front Immunol, 2019, 10: 2852. DOI: 10.3389/fimmu.2019.02852.

BILZER M, ROGGEL F, GERBES A L. Role of Kupffer cells in host defense and liver disease[J]. Liver Int, 2006, 26(10): 1175-1186. DOI: 10.1111/j.1478-3231.2006.01342.x.

QIU P, LIU Y, ZHANG J. Review: the role and mechanisms of macrophage autophagy in sepsis[J]. Inflammation, 2019, 42(1): 6-19. DOI: 10.1007/s10753-018-0890-8.

XIE J F, WANG H L, KANG Y, et al . The epidemiology of sepsis in Chinese ICUs: a national cross-sectional survey[J ] . Crit Care Med, 2020, 48(3): e209-e218. DOI: 10.1097/CCM.0000000000004155.

ZHOU Z, XU M J, GAO B. Hepatocytes: a key cell type for innate immunity[J]. Cell Mol Immunol, 2016, 13(3): 301-315. DOI: 10.1038/cmi.2015.97.

NORRIS C A, HE M, KANG L I, et al . Synthesis of IL-6 by hepatocytes is a normal response to common hepatic stimuli[J ] . PLoS One, 2014, 9(4): e96053. DOI: 10.1371/journal.pone.0096053.

DAVIES S P, TERRY L V, WILKINSON A L, et al . Cell-in-cell structures in the liver: a tale of four E's[J ] . Front Immunol, 2020, 11: 650. DOI: 10.3389/fimmu.2020.00650.

SEEBERG J C, LOIBL M, MOSER F, et al . Non-professional phagocytosis: a general feature of normal tissue cells[J ] . Sci Rep, 2019, 9(1): 11875. DOI: 10.1038/s41598-019-48370-3.

SCHWEGLER M, WIRSING A M, DOLLINGER A J, et al . Clearance of primary necrotic cells by non-professional phagocytes[J ] . Biol Cell, 2015, 107(10): 372-387. DOI: 10.1111/boc.201400090.

DAVIES S P, REYNOLDS G M, STAMATAKI Z. Clearance of apoptotic cells by tissue epithelia: a putative role for hepatocytes in liver efferocytosis[J]. Front Immunol, 2018, 9: 44. DOI: 10.3389/fimmu.2018.00044.

NIÑO D F, CAUVI D M, DE MAIO A. Itraconazole, a commonly used antifungal, inhibits Fcγ receptor-mediated phagocytosis: alteration of Fcγ receptor glycosylation and gene expression[J]. Shock, 2014, 42(1): 52-59. DOI: 10.1097/SHK.0000000000000169.

MATUSZEWSKA M, CIESLIK M, WILKANIEC A, et al . The role of bromodomain and extraterminal (BET) proteins in controlling the phagocytic activity of microglia in vitro : relevance to alzheimer's disease[J ] . Int J Mol Sci, 2022, 24(1): 13. DOI: 10.3390/ijms24010013..

CARPENTIER K S, DAVENPORT B J, HAIST K C, et al . Discrete viral E2 lysine residues and scavenger receptor MARCO are required for clearance of circulating alpha viruses[J ] . eLife, 2019, 8: 49163. DOI: 10.7554/elife.49163.

XING Q Q, FENG Y X, SUN H M, et al . Scavenger receptor MARCO contributes to macrophage phagocytosis and clearance of tumor cells[J ] . Exp Cell Res, 2021, 408(2): 112862. DOI: 10.1016/j.yexcr.2021.112862.

ZHANG J M, ZHU J P, BU X, et al . Cdc42 and RhoB activation are required for mannose receptor-mediated phagocytosis by human alveolar macrophages[J ] . Mol Biol Cell, 2005, 16(2): 824-834. DOI: 10.1091/mbc.e04-06-0463.

O'CALLAGHAN A A, DEMPSEY E, IYER N, et al . Intestinal metabolites influence macrophage phagocytosis and clearance of bacterial infection[J ] . Front Cell Infect Microbiol, 2021, 11: 622491. DOI: 10.3389/fcimb.2021.622491.

MANGOLD C A, MASSER D R, STANFORD D R, et al . CNS-wide sexually dimorphic induction of the major histocompatibility complex 1 pathway with aging[J ] . J Gerontol A Biol Sci Med Sci, 2017, 72(1): 16-29. DOI: 10.1093/gerona/glv232.

EDER C. Mechanisms of interleukin-1beta release[J]. Immunobiology, 2009, 214(7): 543-553. DOI: 10.1016/j.imbio.2008.11.007.

BODE J G, ALBRECHT U, HÄUSSINGER D, et al . Hepatic acute phase proteins—Regulation by IL-6- and IL-1-type cytokines involving STAT3 and its crosstalk with NF-κB-dependent signaling[J ] . Eur J Cell Biol, 2012, 91(6/7): 496-505. DOI: 10.1016/j.ejcb.2011.09.008.

LI P Z, HE K, LI J Z, et al . The role of Kupffer cells in hepatic diseases[J ] . Mol Immunol, 2017, 85: 222-229. DOI: 10.1016/j.molimm.2017.02.018.

SCHUMANN J. It is all about fluidity: fatty acids and macrophage phagocytosis[J]. Eur J Pharmacol, 2016, 785: 18-23. DOI: 10.1016/j.ejphar.2015.04.057.

VEGA F M, RIDLEY A J. The RhoB small GTPase in physiology and disease[J]. Small Gtpases, 2018, 9(5): 384-393. DOI: 10.1080/21541248.2016.1253528.

MIAO C H, YU M Y, PEI G, et al . An infection-induced RhoB-Beclin 1-Hsp90 complex enhances clearance of uropathogenic Escherichia coli [J ] . Nat Commun, 2021, 12(1): 2587. DOI: 10.1038/s41467-021-22726-8..

YOSHIDA T, DAS N A, CARPENTER A J, et al . Minocycline reverses IL-17A/TRAF 3 IP 2 -mediated p38 MAPK/NF-κB/iNOS/NO-dependent cardiomyocyte contractile depression and death[J ] . Cell Signal, 2020, 73: 109690. DOI: 10.1016/j.cellsig.2020.109690..

CAPUTA G, MATSUSHITA M, SANIN D E, et al . Intracellular infection and immune system cues rewire adipocytes to acquire immune function[J ] . Cell Metab, 2022, 34(5): 747-760. e6. DOI: 10.1016/j.cmet.2022.04.008.

CHEN H I, HU W S, HUNG M Y, et al . Protective effects of luteolin against oxidative stress and mitochondrial dysfunction i n endothelial cells[J ] . Nutr Metab Cardiovasc Dis, 2020, 30(6): 1032-1043. DOI: 10.1016/j.numecd.2020.02.014.

NAUSEEF W M. The phagocyte NOX2 NADPH oxidase in microbial killing and cell signaling[J]. Curr Opin Immunol, 2019, 60: 130-140. DOI: 10.1016/j.coi.2019.05.006.

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