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Fgl2对小鼠急性肾损伤中巨噬细胞极化的作用
Role of Fgl2 in macrophage polarization during acute kidney injury in mice
- 陆军军医大学学报 2024年46卷第13期 页码：1467-1476
- 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：
[彭侃夫，E-mail: 392906786@qq.com PENG Kanfu, E-mail: 392906786@qq.com
- 基金信息：国家自然科学基金面上项目(8187032042);重庆市自然科学基金面上项目(CSTB2023NSCQ-MSX0570)
DOI：10.16016/j.2097-0927.202401019
中图分类号： R363.21;R392.12;R692收稿：2024-01-04，
修回：2024-04-26，
纸质出版：2024-07-15
- 稿件说明：
移动端阅览
朱琳, 许桂莲, 谢攀, 等. Fgl2对小鼠急性肾损伤中巨噬细胞极化的作用[J]. 陆军军医大学学报, 2024,46(13):1467-1476.
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.
朱琳, 许桂莲, 谢攀, 等. Fgl2对小鼠急性肾损伤中巨噬细胞极化的作用[J]. 陆军军医大学学报, 2024,46(13):1467-1476. DOI： 10.16016/j.2097-0927.202401019.
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
2
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
2
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(
n
=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
2
与Fgl2
+/+
Saline组比较，Fgl2
+/+
Cis组小鼠肾功能显著下降(
P
＜0.05)，肾小管损伤病理评分显著升高(
P
＜0.05)，肾组织Fgl2表达显著上调(
P
＜0.05)；与Fgl2
+/+
Cis组相比，Fgl2
-/-
Cis组小鼠肾功能显著下降(
P
＜0.05)
，肾损伤相关分子Kim-1、NGAL表达显著升高(
P
＜0.05)，肾小管损伤病理评分显著升高(
P
＜0.05)，巨噬细胞浸润显著增加(
P
＜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(
P
＜0.05)，流式细胞术结果显示M1型巨噬细胞百分比显著升高(
P
＜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(
P
＜0.05)。
结论
2
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
2
To investigate the role of fibrinogen-like protein 2 (Fgl2) in macrophage polarization during cisplatin (Cis)-induced acute kidney injury (Cis-AKI).
Methods
2
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 (
n
=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
2
Compared with the Fgl2
+/+
Saline group
the Fgl2
+/+
Cis group exhibited a significant decline in renal function (
P
<
0.05)
a notable increase in pathological score of renal tubular injury (
P
<
0.05)
and an obvious upregulation of renal tissue Fgl2 expression (
P
<
0.05). Compared with the Fgl2
+/+
Cis group
the Fgl2
-/-
Cis group demonstrated a significant decline in renal function (
P
<
0.05)
an elevation in the expression of renal injury-associated molecules Kim-1 and NGAL (
P
<
0.05)
an increase in pathological score of renal tubular injury (
P
<
0.05)
increase in macrophage infiltration (
P
<
0.05)
and an upregulation in the expression of M1-type macrophage-related molecules IL-6
IL-1β
TNF-α and iNOS (
P
<
0.05)
as well as obviously increase in the percentage of M1-type macrophages as indicated by flow cytometry (
P
<
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 (
P
<
0.05).
Conclusion
2
Fgl2 gene knockout exacerbates Cis-AKI by promoting macrophage polarization towards the M1 phenotype.
关键词
Keywords
references
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