黄芪桂枝五物汤调节M1/M2巨噬细胞改善炎症反应的分子机制研究

王馨慧; 王苏童; 吕穆杰; 田子芳; 马度芳; 王永成; 边莉; 李晓; 姜萍

doi:10.3969/j.issn.1006-2157.2023.06.010

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黄芪桂枝五物汤调节M1/M2巨噬细胞改善炎症反应的分子机制研究

实验研究 | 更新时间：2023-07-05

- 黄芪桂枝五物汤调节M1/M2巨噬细胞改善炎症反应的分子机制研究
- Molecular mechanism of Huangqi Guizhi Wuwu Decoction in regulating M1/M2 macrophages to improve inflammatory response
- 北京中医药大学学报 2023年46卷第6期页码：801-810
- 作者机构：
  
  1.山东中医药大学中医学院　济南　250014
  2.山东中医药大学附属医院
  3.山东省肿瘤防治研究院
- 作者简介：
  
  王馨慧，女，在读硕士生
  # 姜萍，女，博士，教授，主任医师，主要研究方向：中医药治疗风湿免疫病的理论、临床及实验研究，E-mail：lmdlmd6617@163.com
- 基金信息：
  
  国家自然科学基金面上项目(81874449);泰山学者岗位建设项目(2018-35);中国博士后科学基金面上项目(2021M702043);山东省自然科学基金青年项目(ZR2020QH307)
- DOI：10.3969/j.issn.1006-2157.2023.06.010
  中图分类号： R285.5
- 纸质出版日期：2023-06-30，
  
  网络出版日期：2023-04-26，
  
  收稿日期：2022-12-28，
- 稿件说明：
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王馨慧, 王苏童, 吕穆杰, 等. 黄芪桂枝五物汤调节M1/M2巨噬细胞改善炎症反应的分子机制研究[J]. 北京中医药大学学报, 2023,46(6):801-810.

WANG Xinhui, WANG Sutong, LYU Mujie, et al. Molecular mechanism of Huangqi Guizhi Wuwu Decoction in regulating M1/M2 macrophages to improve inflammatory response[J]. Journal of Beijing University of Traditional Chinese Medicine, 2023,46(6):801-810.
王馨慧, 王苏童, 吕穆杰, 等. 黄芪桂枝五物汤调节M1/M2巨噬细胞改善炎症反应的分子机制研究[J]. 北京中医药大学学报, 2023,46(6):801-810. DOI： 10.3969/j.issn.1006-2157.2023.06.010.

WANG Xinhui, WANG Sutong, LYU Mujie, et al. Molecular mechanism of Huangqi Guizhi Wuwu Decoction in regulating M1/M2 macrophages to improve inflammatory response[J]. Journal of Beijing University of Traditional Chinese Medicine, 2023,46(6):801-810. DOI： 10.3969/j.issn.1006-2157.2023.06.010.

摘要

目的

探讨黄芪桂枝五物汤调节M1/M2巨噬细胞极化的机制。

方法

U937细胞经脂多糖诱导为M1巨噬细胞，建立α7烟碱型乙酰胆碱受体(α7nAchR)沉默和激动模型，实验分为对照组、10%含药血清组、siRNA阴性对照(NC)10%含药血清组、siRNA α7nAchR10%含药血清组和α7nAchR激动剂GTS-21组。免疫荧光检测M1、M2型巨噬细胞数量，定量反转录PCR检测M1、M2型巨噬细胞标志基因mRNA表达水平，酶联免疫吸附测定检测M1型巨噬细胞产生的促炎因子和M2型巨噬细胞产生的抗炎因子水平，蛋白质印迹法检测巨噬细胞α7nAchR表达。

结果

与对照组相比，10%含药血清组M1型巨噬细胞数量减少(

0.01)，M2型巨噬细胞数量增加(

0.01)；M1型巨噬细胞基因黏附蛋白白细胞整合素(Cd11c)、诱导型一氧化氮合酶(iNOS)表达均降低(

0.01)，M2型巨噬细胞基因精氨酸酶1(Arg1)、甘露糖受体1(CD206)表达均升高(

0.01)；M1型巨噬细胞产生的促炎因子白细胞介素(IL)-1β、IL-6含量减少(

0.01)，M2型巨噬细胞产生的抗炎因子IL-10和转化生长因子－β(TGF-β)含量升高(

0.01)；10%含药血清组、siRNA NC 10%含药血清组、GTS-21组α7nAchR表达均增加(P

0.01)。与siRNA α7nAchR 10%含药血清组比较，siRNA NC 10%含药血清组、GTS-21组M1型巨噬细胞数量减少(

0.01)，M2型巨噬细胞数量增加(

0.01)；siRNA NC 10%含药血清组Cd11c、iNOS表达降低(

0.01)，Arg1表达升高(

0.01)；siRNA NC 10%含药血清组IL-1β、IL-6含量减少(

0.01)，IL-10、TGF-β含量升高(

0.01)；siRNA NC 10%含药血清组、GTS-21组α7nAchR表达均增加(

0.05，

0.01)。

结论

黄芪桂枝五物汤可能通过激动α7nAchR促使M1/M2型巨噬细胞极化，改善炎症反应。

Abstract

Objective

We aimed to investigate the mechanism of

Huangqi Guizhi Wuwu

Decoction in regulating M1/M2 macrophage polarization.

Methods

U937 cells induced by lipopolysaccharide into M1 macrophages

established α7 nicotinic acetylcholine receptor (α7nAchR) silencing and agonist model. There were divided into the control group

the 10% medicated serum group

the siRNA NC 10% medicated serum group

the siRNA α7nAchR 10% medicated serum group

and the GTS-21 group acting as an α7nAchR agonist. Immunofluorescence was used to detect the number of M1 and M2 macrophages

quantitative real-time PCR was used to detect the mRNA levels of M1 and M2 macrophage marker genes

pro-inflammatory factors produced by M1 macrophages and anti-inflammatory factors produced by M2 macrophages were measured by ELISA. Western blotting was used to detect the expression levels of α7nAchR in macrophages.

Results

Compared with the control group

the number of M1 macrophages decreased (

0.01) and the number of M2 macrophages increased (

0.01) in the 10% medicated serum group. The expression of the M1 macrophage genes adhesion protein leukocyte integrin (Cd11c) and inducible nitric oxide synthase (iNOS) was reduced (

0.01)

while expression of the M2 macrophage genes arginase 1 (Arg1) and mannose receptor 1 (CD206) increased (

0.01). The levels of the pro-inflammatory factors interleukin-1β (IL-1β) and interleukin-6 (IL-6) produced by M1 macrophages decreased (

0.01)

and the levels of the anti-inflammatory factors interleukin-10 (IL-10) and transforming growth factor-β (TGF-β) produced by M2 macrophages increased (

0.01). Furthermore

α7nAchR expression increased in the 10% medicated serum group

the siRNA NC 10% medicated serum group

and the GTS-21 group (

0.01). Compared with the siRNA α7nAchR 10% medicated serum group

the number of M1 macrophages decreased (

0.01)

and the number of M2 macrophages increased (

0.01) in the siRNA NC 10% medicated serum group and GTS-21 group. The expression of Cd11c and iNOS decreased (

0.01) and the expression of Arg1 increased (

0.01) in the siRNA NC 10% medicated serum group

and the levels of IL-1β and IL-6 were reduced (

0.01) in the siRNA NC 10% medicated serum group. The levels of IL-10 and TGF-β increased (

0.01)

and the expression of α7nAchR increased in the siRNA NC 10% medicated serum group and GTS-21 group (

0.05

0.01).

Conclusion

Huangqi Guizhi Wuwu

Decoction may improve the inflammatory response by activating α7nAchR to induce M1/M2 macrophage polarization.

关键词

α7烟碱型乙酰胆碱受体巨噬细胞极化炎症黄芪桂枝五物汤小鼠

Keywords

α7nAchRmacrophagespolarizationinflammationHuangqi Guizhi Wuwu Decoctionmice

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