Role of Fgl2 in macrophage polarization during acute kidney injury in mice

Lin ZHU; Guilian XU; Pan XIE; Kanfu PENG

doi:10.16016/j.2097-0927.202401019

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Role of Fgl2 in macrophage polarization during acute kidney injury in mice

Basic Medicine | 更新时间：2026-01-09

- Role of Fgl2 in macrophage polarization during acute kidney injury in mice
- Journal of Army Medical University Vol. 46, Issue 13, Pages: 1467-1476(2024)
- 作者机构：
  
  1.400038 重庆，陆军军医大学(第三军医大学)第一附属医院肾内科
  2.400038 重庆，陆军军医大学(第三军医大学)基础医学院免疫学教研室
- 作者简介：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(8187032042);the General Program of Natural Science Foundation of Chongqing(CSTB2023NSCQ-MSX0570)
- DOI：10.16016/j.2097-0927.202401019
  CLC： R363.21;R392.12;R692
- Received：04 January 2024，
  
  Revised：2024-04-26，
  
  Published：15 July 2024
- 稿件说明：
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Lin ZHU, Guilian XU, Pan XIE, et al. Role of Fgl2 in macrophage polarization during acute kidney injury in mice[J]. Journal of Army Medical University, 2024, 46(13): 1467-1476.
DOI：

Lin ZHU, Guilian XU, Pan XIE, et al. Role of Fgl2 in macrophage polarization during acute kidney injury in mice[J]. Journal of Army Medical University, 2024, 46(13): 1467-1476. DOI： 10.16016/j.2097-0927.202401019.

摘要

目的

探讨纤维蛋白原样蛋白2(fibrinogen-like protein2，Fgl2)对顺铂诱导的急性肾损伤(cisplatin-induced acute kidney injury，Cis-AKI)中巨噬细胞极化的作用。

方法

选取24只8~10周龄(体质量20~25 g)的雄性野生型(Fgl2

+/+

)小鼠和Fgl2基因敲除(Fgl2

-/-

)小鼠各12只。单次腹腔注射生理盐水(saline)或顺铂(cisplatin，Cis)处理3 d，按随机数字表法分成4组(

=6)：Fgl2

+/+

Saline组、Fgl2

+/+

Cis组、Fgl2

-/-

Saline组和Fgl2

-/-

Cis组。3 d后，检测肾功能相关指标血肌酐(serum creatinine，Scr)、尿素氮(blood urea nitrogen，BUN)的水平，HE染色评估肾损伤程度；Western blot检测肾组织中Fgl2和肾损伤分子1(kidney injury molecule 1，Kim-1)的表达；RT-qPCR检测Fgl2、Kim-1、中性粒细胞明胶酶相关脂质运载蛋白(neutrophil gelatinase-associated lipocalin

NGAL)、IL-6、IL-12p40、IL-1β、诱导型一氧化氮合酶(inducible nitric oxide synthase，iNOS)、TNF-α的表达；免疫组化检测肾脏Fgl2和巨噬细胞(F4/80

)的表达；免疫荧光检测肾组织中巨噬细胞(F4/80

)和M1型巨噬细胞(F4/80

CD86

)的表达；流式细胞术检测肾组织中巨噬细胞(F4/80

)及M1型巨噬细胞(F4/80

MHC Ⅱ

)、M2型巨噬细胞(F4/80

CD206

)的表达。

结果

与Fgl2

+/+

Saline组比较，Fgl2

+/+

Cis组小鼠肾功能显著下降(

＜0.05)，肾小管损伤病理评分显著升高(

＜0.05)，肾组织Fgl2表达显著上调(

＜0.05)；与Fgl2

+/+

Cis组相比，Fgl2

-/-

Cis组小鼠肾功能显著下降(

＜0.05)

，肾损伤相关分子Kim-1、NGAL表达显著升高(

＜0.05)，肾小管损伤病理评分显著升高(

＜0.05)，巨噬细胞浸润显著增加(

＜0.05)，M1型巨噬细胞相关分子IL-6、IL-1β、TNF-α和iNOS表达显著上升(

＜0.05)，流式细胞术结果显示M1型巨噬细胞百分比显著升高(

＜0.05)，M2型巨噬细胞百分比无明显变化，且M1型占比显著高于M2型(

＜0.05)。

结论

Fgl2基因敲除可通过促进巨噬细胞向M1型转化加重Cis-AKI。

Abstract

Objective

To investigate the role of fibrinogen-like protein 2 (Fgl2) in macrophage polarization during cisplatin (Cis)-induced acute kidney injury (Cis-AKI).

Methods

Twelve male wild-type (Fgl2

+/+

) mice and 12 Fgl2 gene knockout (Fgl2

-/-

) mice

aged 8~10 weeks and weighing 20~25 g

were selected

and then after being administered with a single intraperitoneal injection of either saline or Cis for 3 d

they were randomly divided into 4 groups (

=6): Fgl2

+/+

Saline group

Fgl2

+/+

Cis group

Fgl2

-/-

Saline group and Fgl2

-/-

Cis group. Kidney function indicators such as serum creatinine (Scr) and blood urea nitrogen (BUN) levels were measured 3 d later

and kidney injury was assessed by HE staining. Western blot analysis was performed to evaluate the expression of Fgl2 and kidney injury molecule 1 (Kim-1) in the renal tissues. RT-qPCR was conducted to assess the expression levels of Fgl2

Kim-1

neutrophil gelatinase-associated lipocalin (NGAL)

IL-6

IL-12p40

IL-1β

inducible nitric oxide synthase (iNOS) and TNF-α in the renal tissues. Immunohistochemical assay was employed to detect the expression of Fgl2 and macrophages (F4/80

) in the kidneys. Immunofluorescence staining was utilized to examine the expression of macrophages (F4/80

) and M1-type macrophages (F4/80

CD86

) in the renal tissues. Flow cytometry was employed to analyze the expression of macrophages (F4/80

) as well as M1

-type macrophages (F4/80

MHC Ⅱ

) and M2-type macrophages (F4/80

CD206

) in the renal tissues.

Results

Compared with the Fgl2

+/+

Saline group

the Fgl2

+/+

Cis group exhibited a significant decline in renal function (

0.05)

a notable increase in pathological score of renal tubular injury (

0.05)

and an obvious upregulation of renal tissue Fgl2 expression (

0.05). Compared with the Fgl2

+/+

Cis group

the Fgl2

-/-

Cis group demonstrated a significant decline in renal function (

0.05)

an elevation in the expression of renal injury-associated molecules Kim-1 and NGAL (

0.05)

an increase in pathological score of renal tubular injury (

0.05)

increase in macrophage infiltration (

0.05)

and an upregulation in the expression of M1-type macrophage-related molecules IL-6

IL-1β

TNF-α and iNOS (

0.05)

as well as obviously increase in the percentage of M1-type macrophages as indicated by flow cytometry (

0.05)

while there was no significant change in the percentage of M2-type macrophages

and the proportion of M1-type macrophages was significantly higher than that of M2-type macrophages (

0.05).

Conclusion

Fgl2 gene knockout exacerbates Cis-AKI by promoting macrophage polarization towards the M1 phenotype.

关键词

Keywords

references

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