Low-dose 5-aminolevulinic acid photodynamic therapy of promotes polarity switch of M1 macrophages

Tianzhen XIAO; Liqun WANG; Anhui WANG; Yang TAN; Jinyi CHEN; Xia LEI

doi:10.16016/j.2097-0927.202207093

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Low-dose 5-aminolevulinic acid photodynamic therapy of promotes polarity switch of M1 macrophages

Basic Medicine | 更新时间：2026-01-09

- Low-dose 5-aminolevulinic acid photodynamic therapy of promotes polarity switch of M1 macrophages
- Journal of Army Medical University Vol. 45, Issue 3, Pages: 219-226(2023)
- 作者机构：
  
  400042 重庆，陆军特色医学中心皮肤科
- 作者简介：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(81773348);the Clinical Research Talent Training Program of Army Medical University(2018XLC3065)
- DOI：10.16016/j.2097-0927.202207093
  CLC： R329.24;R392.12;R454.2
- Received：13 July 2022，
  
  Revised：2022-10-09，
  
  Published：15 February 2023
- 稿件说明：
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Tianzhen XIAO, Liqun WANG, Anhui WANG, et al. Low-dose 5-aminolevulinic acid photodynamic therapy of promotes polarity switch of M1 macrophages[J]. Journal of Army Medical University, 2023, 45(3): 219-226.
DOI：

Tianzhen XIAO, Liqun WANG, Anhui WANG, et al. Low-dose 5-aminolevulinic acid photodynamic therapy of promotes polarity switch of M1 macrophages[J]. Journal of Army Medical University, 2023, 45(3): 219-226. DOI： 10.16016/j.2097-0927.202207093.

摘要

目的

观察低剂量5-氨基酮戊酸光动力疗法(5-aminolevulinic acid photodynamic therapy

ALA-PDT)对M1型巨噬细胞极性转换的影响并初步探索其机制。

方法

M1组用含100 ng/mL脂多糖(LPS)的培养基培养RAW264.7细胞24 h

诱导其成为M1型巨噬细胞

对照组用不含LPS的培养基培养细胞24 h。CCK-8法检测不同光照强度的低剂量ALA-PDT疗法对M1巨噬细胞的毒性作用和对M1型标志物诱导型一氧化氮合酶(iNOS)表达的影响; 将M1型巨噬细胞分为4组: M1组正常培养

ALA组仅孵育ALA

红光组仅照射红光

ALA-PDT组孵育ALA后进行红光照射。采用实时荧光定量PCR检测M1型标志物iNOS和白介素6(IL-6)、M2型标志物白介素10(IL-10)和精氨酸酶1(arginase 1

Arg-1) mRNA表达水平

用ELISA法检测IL-6、IL-10的分泌水平

Western blot检测iNOS、M1型标志物白介素1 β(IL-1β)、Arg-1及自噬相关的蛋白泛素结合蛋白P62、微管相关蛋白1轻链3(LC3)的表达。

结果

与对照组比较

M1组iNOS和IL-6 mRNA表达升高(

0.05)

iNOS、IL-1β蛋白表达升高(

0.05)

Arg-1的表达降低(

0.05)

IL-6的分泌水平升高(

0.05)。随着光照强度的增强

ALA-PDT疗法对M1型巨

噬细胞的毒性作用逐渐增强(

0.05)

光照强度大于2 J/cm

时

对iNOS的表达有抑制作用。与M1组、ALA组和红光组比较

ALA-PDT组Arg-1和IL-10的mRNA表达升高、iNOS的mRNA表达降低(

0.05)

Arg-1的蛋白表达升高、iNOS和IL-1β的蛋白表达降低(

0.05)

IL-10的分泌水平升高(

0.05)

IL-6的分泌水平降低(

0.05)

P62表达降低(

0.05)

LC3Ⅱ/LC3Ⅰ的比值升高(

0.05)。

结论

低剂量ALA-PDT疗法可促进M1型巨噬细胞转换为M2型巨噬细胞

其作用机制可能与自噬相关。

Abstract

Objective

To determine the effect of low-dose 5-aminolevulinic acid photodynamic therapy (ALA-PDT) on the polarity switch of M1 macrophages and explore its mechanism primarily.

Methods

RAW264.7 cells were cultured with 100 ng/mL lipopolysaccharide (LPS) for 24 h to induce M1 macrophages.The control group was cultured in the medium without LPS for 24 h.CCK-8 assay was used to detect the toxic effect of low-dose ALA-PDT therapy with different light doses on M1 macrophages

and the expression of M1 type marker

inducible nitric oxide synthase (iNOS) was measured with Western blotting.Then M1 macrophages were divided into M1 group

ALA group

red light group and ALA-PDT group (ALA treatment followed by red light irradiation).The mRNA expression levels of inducible M1 type markers iNOS and interleukin 6(IL-6)

and M2 type markers interleukin 10(IL-10) and arginase 1(Arg-1) were detected by real-time fluorescent quantitative PCR

and the secretions of IL-6 and IL-10 were detected by ELISA.Western blotting was applied to detect the protein levels of iNOS

interleukin 1β(IL-1β)

Arg-1

and autophagy-related proteins

ubiquitin-binding protein P62 and tubule-associated protein 1 light chain 3(LC3).

Results

Compared with the control group

the mRNA levels of iNOS and IL-6 were higher (

0.05)

the protein levels of iNOS and IL-1β were higher (

0.05)

and that of Arg-1 was lower (

0.05)

and the secretion level of IL-6 was higher in the M1 group (

0.05).With the increase of light intensity

the toxic effect of ALA-PDT therapy on M1 macrophages was gradually increased (

0.05)

and when the intensity higher than 2 J/cm

the expression of iNOS was inhibited.The mRNA levels of Arg-1 and IL-10 were increased

while that of iNOS was decreased (

0.05)

the protein expression of Arg-1 was elevated while those of iNOS and IL-1β were decreased (

0.05)

the secretion level of IL-10 was increased (

0.05) but that of IL-6 decreased (

0.05)

and the protein level of P62 was decreased (

0.05) and the ratio of LC3Ⅱ/LC3Ⅰ was increased (

0.05) in the ALA-PDT group when compared with the M1 group

ALA group and red light group.

Conclusion

Low-dose ALA-PDT therapy can promote the conversion of M1-type macrophages to M2-type macrophages

and its mechanism may be related to autophagy.

关键词

Keywords

references

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