Research on Regulatory Mechanism of Verbenalin on HCoV-229E-infected Macrophage Injury Based on Mitophagy
10.13422/j.cnki.syfjx.20251240
- VernacularTitle:基于线粒体自噬探讨马鞭草苷对HCoV-229E感染巨噬细胞损伤的调控机制
- Author:
Qiyue SUN
1
;
Lei BAO
1
;
Zihan GENG
1
;
Ronghua ZHAO
1
;
Shuran LI
1
;
Xihe CUI
1
;
Jingsheng ZHANG
1
;
Xian LIU
1
;
Rui XIE
1
;
Xiaolan CUI
1
;
Shanshan GUO
1
;
Jing SUN
1
Author Information
1. Institute of Chinese Materia Medica， China Academy of Chinese Medical Sciences， Beijing 100029， China
- Publication Type:Journal Article
- Keywords:
verbenalin;
human coronavirus （HCoV）-229E;
mononuclear macrophage leukemia cell （RAW264.7）;
mitophagy;
mitochondrial membrane potential
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2025;31(21):29-37
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo investigate the protective effect and mechanism of verbenalin on mouse mononuclear macrophage leukemia cells （RAW264.7） damaged by human coronavirus （HCoV）-229E infection， thereby providing experimental evidence for its development and application. MethodsRAW264.7 macrophages were infected with different concentrations of HCoV-229E to establish a coronavirus-induced macrophage injury model using the cell counting kit-8 （CCK-8） assay for assessing cell proliferation and viability. Cells were randomly divided into four groups： normal control， verbenalin group （125 μmol·L-1）， model group （HCoV-229E）， and HCoV-229E + verbenalin group （HCoV-229E + 125 μmol·L-1 verbenalin）. Cell viability was measured using the CCK-8 assay， and the maximum non-toxic concentration （CC0）， half-maximal cytotoxic concentration （CC50）， half-maximal effective concentration （EC50）， and selectivity index （SI） of verbenalin were calculated. Calcein/PI double staining was used to assess cell viability and cytotoxicity， and JC-1 staining was applied to evaluate changes in mitochondrial membrane potential （MMP）. mito-Keima adenovirus labeling was used to assess mitophagy levels in each group. ResultsA macrophage infection model was successfully established by infecting RAW264.7 cells with the original concentration of HCoV-229E for 36 h. The CC0 of verbenalin was 125 μmol·L-1. The CC50 was 448.25 μmol·L-1. The EC50 against HCoV-229E-infected cells was 46.28 μmol·L-1， and the SI was 9.68. Compared with the normal group， the model group showed significantly reduced cell survival rate （P<0.01）， increased cell death rate （P<0.01）， decreased MMP （P<0.01）， and suppressed mitophagy （P<0.01）. In contrast， verbenalin treatment significantly improved cell survival rate （P<0.01）， reduced cell death rate （P<0.01）， alleviated MMP loss （P<0.01）， and enhanced mitophagy levels （P<0.01） compared with the model group. ConclusionVerbenalin can enhance the survival rate of macrophages following HCoV-229E infection. The underlying mechanism may be associated with the activation of mitophagy， maintenance of MMP stability， and alleviation of mitochondrial damage.
