CX3CR1 silencing-mediated macrophage polarization attenuates CCl<sub>4</sub> -induced liver fibrosis in mice

Shasha HUANG; Yanli ZOU; Jie TAN; Meng LIU; Wei CHEN

doi:10.16016/j.2097-0927.202111205

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CX3CR1 silencing-mediated macrophage polarization attenuates CCl₄ -induced liver fibrosis in mice

Basic Medicine | 更新时间：2026-02-13

- CX3CR1 silencing-mediated macrophage polarization attenuates CCl₄ -induced liver fibrosis in mice
- Journal of Army Medical University Vol. 44, Issue 13, Pages: 1314-1321(2022)
- 作者机构：
  
  430074 武汉，武汉大学附属同仁医院(武汉市第三医院)消化内科
- 作者简介：
- 基金信息：
  
  the Scientific Research Project of Wuhan Health and Family Planning Commission(WX18Q12)
- DOI：10.16016/j.2097-0927.202111205
  CLC： R322.47;R329.24;R392.2
- Received：28 November 2021，
  
  Revised：2022-03-01，
  
  Published：15 July 2022
- 稿件说明：
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Shasha HUANG, Yanli ZOU, Jie TAN, et al. CX3CR1 silencing-mediated macrophage polarization attenuates CCl₄ -induced liver fibrosis in mice[J]. Journal of Army Medical University, 2022, 44(13): 1314-1321.
DOI：

Shasha HUANG, Yanli ZOU, Jie TAN, et al. CX3CR1 silencing-mediated macrophage polarization attenuates CCl₄ -induced liver fibrosis in mice[J]. Journal of Army Medical University, 2022, 44(13): 1314-1321. DOI： 10.16016/j.2097-0927.202111205.

摘要

目的

探讨CX3CR1在CCl

诱导的小鼠肝纤维化中的作用。

方法

将小鼠分为正常组(

=24)、CCl

组(

=36)、CCl

+sh-NC组(

=36)、CCl

+sh-CX3CR1组(

=36)。除正常组外，其余组通过腹腔注射CCl

诱导肝纤维化。从CCl

诱导的肝纤维化4周开始，每周分别向CCl

+sh-NC组和CCl

+sh-CX3CR1组小鼠尾静脉注射sh-NC或sh-CX3CR1，持续2周。通过免疫组织化学确定小鼠肝脏组织中CX3CR1的表达情况，免疫荧光染色分析肝脏组织中M1-巨噬细胞(F4/80

iNOS

)或M2-巨噬细胞(F4/80

CD206

)表达。ELISA试剂盒法检测小鼠血清IL-1β、IL-6和TGF-β1水平。使用流式细胞仪测定原代肝脏巨噬细胞与活化HSC共培养后巨噬细胞M1和M2表型变化。

结果

在健康的非纤维化对照肝脏中几乎检测不到CX3CR1，而在CCl

处理4周和6周后，CX3CR1强烈上调。通过沉默CCl

小鼠CX3CR1表达，小鼠肝炎症细胞募集和纤维组织沉积化的严重程度均明显减轻，并且α-SMA、N-cadherin和Vimentin的蛋白水平降低，E-cadherin的蛋白表达增加。与CCl

+sh-NC组相比，CCl

+sh-CX3CR1组小鼠血清IL-1β、IL-6和TGF-β1水平均下调4.6~8.1倍(

0.01)，且M2-巨噬细胞的百分率和M2/M1比值均显著增加(

0.01)。在用CX3CR1沉默的小鼠肝星状细胞和巨噬细胞的共培养中，高M2/M1-巨噬细胞比值抑制胶原的产生。

结论

CX3CR1沉默通过促进M1向M2-巨噬细胞的极化发挥抗肝纤维化作用。

Abstract

Objective

To investigate the role of CX3CR1 in liver fibrosis induced by CCl

in mice.

Methods

The mice were divided into normal group (

=24)

CCl

group (

=36)

CCl

+sh-NC group (

=36)

and CCl

+sh-CX3CR1 group (

=36)

and all the groups were induced liver fibrosis by intraperitoneal injection of CCl

except the normal group. From the 4th week of CCl

injection

the mice of CCl

+sh-NC group and CCl

+sh-CX3CR1 group were respectively injected with shRNA or shRNA-CX3CR1 by tail vein weekly

for 2 weeks. The expression of CX3CR1 in the liver tissues was determined by immunohistochemistry

and the number of M1 macrophages (F4/80

iNOS

) or M2 macrophages (F4/80

CD206

) in the liver tissues was measured by immunofluorescence staining. ELISA was used to detect the serum levels of IL-1β

IL-6 and TGF-β1 in mice

and flow cytometry was adopted to analyze the phenotypic changes of M1 and M2 macrophages after co-culture of primary liver macrophages with activated hepatic stellate cells (HSC).

Results

CX3CR1 was hardly detected in healthy non-fibrotic control livers

but strongly up-regulated 4~6 weeks after CCl

treatment. By silencing CX3CR1 expression in CCl

-induced liver fibrosis mice

the severity of inflammatory cell recruitment and fibrous tissue deposition in liver tissues was significantly reduced

and the protein levels of α-SMA

N-cadherin and Vimentin were decreased

while the expression of E-cadherin increased. As compared with the CCl

+sh-NC group

the serum levels of IL-1β

IL-6 and TGF-β1 were down-regulated by 4.6-8.1 times in the CCl

+sh-CX3CR1 group (

0.01)

and the percentage of M2-macrophages as well as the M2/M1 ratio were remarkably elevated (

0.01). In the co-cultures of HSC and macrophages extracted from CX3CR1 silencing mice

collagen production was inhibited due to the high M2/M1 ratio.

Conclusion

CX3CR1 silencing plays an anti-hepatic fibrosis role by promoting polarization of M1 to M2 macrophage.

关键词

Keywords

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⁰

CX3CR1 silencing-mediated macrophage polarization attenuates CCl4 -induced liver fibrosis in mice

DOI：10.16016/j.2097-0927.202111205

摘要

Abstract

关键词

Keywords

references

CX3CR1 silencing-mediated macrophage polarization attenuates CCl₄ -induced liver fibrosis in mice