Role of heme oxygenase-1/mitochondrial signaling pathway in mitigation of endotoxin-induced lung injury by mesenchymal stem cell-derived exosomes in alveolar macrophages of mice
10.3760/cma.j.cn131073-20240811-00417
- VernacularTitle:肺泡巨噬细胞HO-1/线粒体信号通路在间充质干细胞来源外泌体减轻小鼠内毒素性肺损伤中的作用
- Author:
Wei CHEN
1
;
Ya WU
1
;
Xiaoyang WU
1
;
Jianbo YU
1
;
Lirong GONG
1
Author Information
1. 天津医科大学附属南开医院麻醉与重症医学科，天津 300100
- Publication Type:Journal Article
- Keywords:
Mesenchymal stem cells;
Exosomes;
Mitochondria;
Heme oxygenase-1;
Endotoxemia;
Macrophages, alveolar
- From:
Chinese Journal of Anesthesiology
2025;45(4):474-481
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To evaluate whether the mechanism by which mesenchymal stem cell-derived exosomes (MSC-exo) mitigated endotoxin-induced lung injury was related to the heme oxygenase-1 (HO-1)/mitochondrial signaling pathway in alveolar macrophages of mice.Methods:In vivo experiment Eighteen C57BL/6 wild-type (WT) mice were divided into 3 groups ( n=6 each) using a random number table method: control group (C group), lipopolysaccharide (LPS) group (L group) and LPS + MSC-exo group (LM group). Six HO-1 conditional knockout mice (HO-1 -/-) were selected and served as HO-1 -/- + MSC-exo + LPS group (HML group). The model of endotoxin-induced lung injury was prepared by injection of LPS 15 mg/kg. MSC-exo (2×10 11 particles) was intravenously injected at 1 h before injection of LPS in LM group. MSC-exo (2×10 11 particles) was intravenously injected and 1 h later LPS was injected in HML group. The expression of HO-1 in macrophages was detected using immunofluorescence, lung injury was assessed following hematoxylin-eosin staining, the wet/dry weight ratio (W/D ratio) was determined, and the mitochondrial morphology was observed with a transmission electron microscope. Cell experiment Alveolar macrophages (MH-S) were divided into 4 groups ( n=20 each) using a random number table method: control group (C group), LPS+ phosphate buffer solution group (LP group), LPS+ MSC-exo group (LM group), and LPS+ MSC-exo+ HO-1 small-interfering RNA group (LMS group). Cells were incubated for 12 h with LPS 10 μg/ml in LP, LM and LMS groups. In addition, LM group was incubated with MSC-exo 100 μg/ml, LP group was incubated with the equal volume of phosphate buffer solution, and the alveolar macrophages were transfected with HO-1 small interfering RNA and incubated with MSC-exo 100 μg/ml in LMS group at the same time. The concentrations of interleukin-1beta (IL-1β) and tumor necrosis factor-alpha (TNF-α) in supernatant were measured by enzyme-linked immunosorbent assay, HO-1 expression was detected by Western blot, the mitochondrial membrane potential was measured using JC-1 staining, and the expression of reactive oxygen species (ROS) was detected by fluorescence. Results:In vivo experiment Compared to C group, the lung injury score and W/D ratio were significantly increased ( P<0.05), the fluorescence signal of HO-1 in macrophages was enhanced, and the damage to mitochondria was aggravated in L group. Compared to L group, the lung injury score and W/D ratio were significantly decreased ( P<0.05), the fluorescence signal of HO-1 in macrophages was enhanced, and the damage to mitochondria was reduced in LM group. Compared to LM group, the lung injury score and W/D ratio were significantly increased ( P<0.05), macrophages had no HO-1 fluorescence signal, and the damage to mitochondria was aggravated in HML group. Cell experiment Compared to C group, the concentrations of IL-1β and TNF-α in supernatant were significantly increased, the expression of HO-1 was up-regulated ( P<0.05), and the mitochondria predominantly exhibited green JC-1 fluorescence, accompanied by an enhanced ROS fluorescence signal in LP group. Compared to LP group, the concentrations of IL-1β and TNF-α in supernatant were significantly decreased, the expression of HO-1 was up-regulated ( P<0.05), and the mitochondria predominantly exhibited red JC-1 fluorescence, accompanied by a weakened ROS fluorescence signal in LM group. Compared to LM group, the concentrations of IL-1β and TNF-α in supernatant were significantly increased, the expression of HO-1 was down-regulated ( P<0.05), and the mitochondria predominantly exhibited green JC-1 fluorescence, accompanied by an enhanced ROS fluorescence signal in LMS group. Conclusions:The mechanism by which MSC-exo attenuates endotoxin-induced lung injury may be related to up-regulation of HO-1 expression in alveolar macrophages and reduction of mitochondrial damage in mice.
