Effects of
Angelica polysaccharide on differentiation and function in M2 macrophages- Journal of Army Medical University Vol. 46, Issue 22, Pages: 2505-2516(2024)
- 作者机构：
261053 山东 潍坊，山东第二医科大学基础医学院，山东省高等学校免疫微环境与炎症性疾病研究特色实验室
- 作者简介：
- 基金信息：the General Program of National Natural Science Foundation of China(81972695);the Introduction and Cultivation Program for Young Innovative Talents in Science and Technology Support Plan of Colleges and Universities in Shandong Province, and the Science and Technology Support Plan for Youth Innovation of Colleges and Universities of Shandong Province(2021KJ106)
DOI：10.16016/j.2097-0927.202405021
CLC： R285.5;R329.28;R331.144Received：08 May 2024，
Revised：2024-08-19，
Published：30 November 2024
- 稿件说明：
移动端阅览
Ke ZHANG, Zhao CUI, Yahan QIN, et al. Effects of
Angelica polysaccharide on differentiation and function in M2 macrophages[J]. Journal of Army Medical University, 2024, 46(22): 2505-2516.Ke ZHANG, Zhao CUI, Yahan QIN, et al. Effects of
Angelica polysaccharide on differentiation and function in M2 macrophages[J]. Journal of Army Medical University, 2024, 46(22): 2505-2516. DOI： 10.16016/j.2097-0927.202405021.
摘要
目的
2
探讨当归多糖(angelica polysaccharide，APS)对M2型巨噬细胞分化、功能的影响及其分子机制。
方法
2
诱导小鼠骨髓来源的巨噬细胞(bone marrow derived macrophage，BMDM)和M2型巨噬细胞，加入APS处理(0、80、160、320 μg/mL)；分离小鼠腹腔巨噬细胞，加入APS处理(0、160 μg/mL)。流式细胞术(flow cytometry
FCM)检测M2型巨噬细胞和腹腔巨噬细胞甘露糖受体(mannose receptor
MR)、CD11b、F4/80、CD163、ARG-1表达水平、凋亡情况以及吞噬能力。根据随机数字法将小鼠分为2组，即APS灌胃组(
n
=5)和对照组(
n
=5)，FCM检测外周血和脾脏中巨噬细胞MR表达水平。荧光显微镜观察BMDM分化的M2型巨噬细胞形态；RT-qPCR检测M2型巨噬细胞MR、ARG-1的mRNA表达水平；免疫荧光检测M2型巨噬细胞分化和功能的分子机制。
结果
2
与0 μg/mL APS组相比，在一定浓度(80~320 μg/mL APS)范围内，随着APS浓度的升高，M2型巨噬细胞的MR表达水平降低(
P
＜0.01)；160 μg/mL APS组腹腔巨噬细胞MR表达水平也降低(
P
＜0.01)。与对照组相比，APS灌胃组小鼠外周血和脾脏中巨噬细胞MR的表达水平也明显下降
(
P
＜0.05)。与0 μg/mL APS组相比，80~320 μg/mL APS组巨噬细胞CD11b、F4/80、CD163表达水平升高(
P
＜0.01)；巨噬细胞形态发生变化，由大多呈梭形且有伪足变为大多为类圆形或不规则且少数细胞有伪足；APS可诱导M2型巨噬细胞凋亡(
P
＜0.05)。与0 μg/mL APS组相比，160 μg/mL APS组M2型巨噬细胞吞噬荧光微球的能力升高(
P
＜0.01)，但ARG-1的表达水平降低(
P
＜0.01)；M2型巨噬细胞MR和ARG-1的mRNA表达降低(
P
＜0.05)；160 μg/mL APS组M2型巨噬细胞的磷酸化的信号转导子和转录激活子6(phosphate acidified-signal transducers and activators of transcription 6
p-STAT6)阳性信号的平均荧光强度明显减低(
P
＜0.05)。
结论
2
APS对M2型巨噬细胞的分化和功能均具有双向调控作用，这可能与APS可下调信号转导子和转录激活子6(signal transducers and activators of transcription 6
STAT6)信号通路相关。
Abstract
Objective
2
To investigate the effect of
Angelica
polysaccharide (APS) on the differentiation and function of M2 macrophages and underlying molecular mechanism.
Methods
2
Mouse bone marrow derived macrophages (BMDM) and M2 macrophages were induced and treated with APS (0
80
160
320 μg/mL); Mouse peritoneal macrophages were isolated and treated with APS (0
160 μg/mL). Flow cytometry (FCM) was used to detect mannose receptor(MR)
CD11b
F4/80
CD163
and ARG-1 expression levels
apoptosis
and phagocytic ability of M2 macrophages and peritoneal macrophages. Mice were randomly divided into APS gavage group and control group
APS was intragastrically administered to mice
and macrophage MR expression level in blood and spleen were detected by FCM. Fluorescence microscopy was used to observe the morphology of BMDM-differentiated M2 macrophages. RT-qPCR was employed to detect the mRNA expression levels of MR and ARG-1 in M2 macrophages. Immunofluorescence assay was performed to detect the expression of the proteins related to molecular mechanism of differentiation and function of M2 macrophages.
Results
2
Compared with the 0 μg/mL APS group
the MR expression level in the M2
macrophages was decreased with the increase of APS concentration within a certain concentration range (80~320 μg/mL)
and the MR expression level in peritoneal macrophages was also decreased in the 160 μg/mL APS treatment group (
P
<
0.01). The expression level of macrophage MR was also significantly decreased in peripheral blood and spleen in the APS gavage mice than the control group (
P
<
0.05). Compared with the 0 μg/mL APS group
the expression levels of CD11b
F4/80
and CD163 in the macrophages were increased in the 80 ~ 320 μg/mL APS treatment groups (
P
<
0.01). The morphology of macrophage had changed
from mostly spindle-shaped and pseudopodia to mostly round or irregular
and even a few cells with pseudopodia. APS induced apoptosis in M2 macrophages (
P
<
0.05). Compared with the 0 μg/mL APS group
M2 macrophages treated with 160 μg/mL APS had an increased ability to phagocytose fluorescent microspheres (
P
<
0.01)
but the expression level of ARG-1 was decreased (
P
<
0.01). The mRNA expression of MR and ARG-1 in M2 macrophages was decreased (
P
<
0.05). The mean fluorescence intensity of phosphate acidified-signal transducers and activators of transcription 6(p-STAT6)-positive signals in M2 macrophages was significantly reduced in the 160 μg/mL APS-treated group (
P
<
0.05).
Conclusion
2
APS has bidirectional regulation on the differentiation and function of M2 macrophages
which may be associated with its downregulation of signal transducers and activators of transcription 6(STAT6) signaling pathway.
关键词
Keywords
references
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