Mechanism of bilobalide promoting neuroprotection of macrophages.
10.19540/j.cnki.cjcmm.20230522.501
- Author:
Yang-Yang CHEN
1
;
Wen-Yuan JU
1
;
Guo-Guo CHU
1
;
Xiao-Hui LI
1
;
Ru-Heng WEI
1
;
Qing WANG
1
;
Bao-Guo XIAO
2
;
Cun-Gen MA
3
Author Information
1. Research Center of Neurobiology/the Key Research Laboratory of Benefiting Qi for Acting Blood Circulation Method to Treat Multiple Sclerosis of State Administration of Traditional Chinese Medicine, Shanxi University of Chinese Medicine Jinzhong 030619, China.
2. Institute of Neurology/Institutes of Brain Science and State Key Laboratory of Medical Neurobiology, Huashan Hospital, Fudan University Shanghai 200025, China.
3. Research Center of Neurobiology/the Key Research Laboratory of Benefiting Qi for Acting Blood Circulation Method to Treat Multiple Sclerosis of State Administration of Traditional Chinese Medicine, Shanxi University of Chinese Medicine Jinzhong 030619, China Institute of Brain Science, Shanxi Datong University Datong 037009, China.
- Publication Type:Journal Article
- Keywords:
bilabolide;
inflammation;
macrophages;
microglia;
neuroprotection;
neurotrophic factor
- MeSH:
Female;
Rats;
Mice;
Animals;
Bilobalides/pharmacology*;
Neuroprotection;
Lipopolysaccharides/toxicity*;
Culture Media, Conditioned/pharmacology*;
Mice, Inbred C57BL;
Macrophages/metabolism*;
Microglia;
Cytokines/metabolism*;
Nerve Growth Factors/pharmacology*;
Inflammation/metabolism*
- From:
China Journal of Chinese Materia Medica
2023;48(15):4201-4207
- CountryChina
- Language:Chinese
-
Abstract:
This study aims to explore the neuroprotective effect of bilobalide(BB) and the mechanisms such as inhibiting inflammatory response in macrophage/microglia, promoting neurotrophic factor secretion, and interfering with the activation and differentiation of peripheral CD4~+ T cells. BB of different concentration(12.5, 25, 50, 100 μg·mL~(-1)) was used to treat the RAW264.7 and BV2 cells for 24 h. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay and cell counting kit-8(CCK-8) were employed to detect the cytotoxicity of BB and appropriate concentration was selected for further experiment. Lipopolysaccharide(LPS) was applied to elicit inflammation in RAW264.7 and BV2 cells, mouse bone marrow-derived macrophages(BMDMs), and primary microglia, respectively. The effect of BB on cell proliferation and secretion of inflammatory cytokines and neurotrophic factors was detected by enzyme-linked immunosorbent assay(ELISA). Spleen monocytes of C57BL/6 female mice(7-8 weeks old) were isolated, and CD4~+ T cells were separated by magnetic beads under sterile conditions. Th17 cells were induced by CD3/CD28 and the conditioned medium for eliciting the inflammation in BMDMs. The content of IL-17 cytokines in the supernatant was detected by ELISA to determine the effect on the activation and differentiation of CD4~+ T cells. In addition, PC12 cells were incubated with the conditioned medium for eliciting inflammation in BMDMs and primary microglia and the count and morphology of cells were observed. The cytoto-xicity was determined by lactate dehydrogenase(LDH) assay. The result showed that BB with the concentration of 12.5-100 μg·mL~(-1) had no toxicity to RAW264.7 and BV2 cells, and had no significant effect on the activity of cell model with low inflammation. The 50 μg·mL~(-1) BB was selected for further experiment, and the results indicated that BB inhibited LPS-induced secretion of inflammatory cytokines. The experiment on CD4~+ T cells showed that the conditioned medium for LPS-induced inflammation in BMDMs promoted the activation and differentiation of CD4~+ T cells, while the conditioned medium of the experimental group with BB intervention reduced the activation and differentiation of CD4~+ T cells. In addition, BB also enhanced the release of neurotrophic factors from BMDMs and primary microglia. The conditioned medium after BB intervention can significantly reduce the death of PC12 neurons, inhibit neuronal damage, and protect neurons. To sum up, BB plays a neuroprotective role by inhibiting macrophage and microglia-mediated inflammatory response and promoting neurotrophic factors.
