Expression and function of α7 nicotinic acetylcholine receptor in thymocytes of myasthenia gravis patients
- VernacularTitle:α7烟碱型乙酰胆碱受体在重症肌无力胸腺细胞中的表达及其功能
- Author:
Yuwei HUANG
1
,
2
,
2
,
3
;
Meng WANG
1
,
2
,
2
,
3
;
Yonghui ZHANG
1
,
2
,
2
,
3
;
Xinzheng CUI
2
,
4
;
Zirui SUN
5
;
Zhiwen ZHANG
2
,
4
;
Chenshuo SHI
2
,
4
;
Qingyong ZHANG
2
,
4
Author Information
1. 1.Myasthenia Gravis Comprehensive Diagnosis and Treatment Center, Henan Provincial People&rsquo
2. s Hospital, Zhengzhou, 450003, P. R. China
3. 2. Comprehensive Laboratory, Henan Provincial People&rsquo
4. Myasthenia Gravis Comprehensive Diagnosis and Treatment Center, Henan Provincial People&rsquo
5. Department of Adult Cardiac Surgery, Fuwai Huazhong Cardiovascular Hospital, Zhengzhou, 450003, P. R. China
- Publication Type:Journal Article
- Keywords:
α7 nicotinic acetylcholine receptor;
myasthenia gravis;
cytokines;
basic research
- From:
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery
2023;30(06):897-902
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the expression of α7 nicotinic acetylcholine receptor (α7 nAChR) in thymocytes of patients with myasthenia gravis (MG) and its effect on cytokine secretion and T cell proliferation. Methods Patients with MG who underwent expanded thoracoscopic thymectomy in the Comprehensive Diagnosis and Treatment Center of the Henan Provincial People’s Hospital from June 2021 to June 2022 were selected and allocated to a MG group. Patients who underwent partial thymectomy to expose the surgical field during the cardiac disease surgery from June 2021 to September 2022 in the Department of Adult Cardiac Surgery of Fuwai Huazhong Cardiovascular Hospital were selected as the control group. Thymic single cell suspensions were prepared from MG and control groups, and the expression of α7 nAChR in thymocytes of the two groups was detected by real-time polymerase chain reaction and Western blotting. Then CD3/CD28 monoclonal antibody coupled with magnetic beads was used to induce T cell activation, and the levels of cytokines interferon-gamma (IFN-γ), tumor necrosis factor-α (TNF-α), interleukin-4 (IL-4), IL-6, IL-10, IL-17, and IL-21 in thymocytes of the two groups were detected by enzyme-linked immunosorbent assay (ELISA). The activated T cells of the MG group were divided into a blank control group, an α7 nAChR antagonist group, and an α7 nAChR agonist group according to different treatment methods. After 72 hours of culture, IFN-γ, TNF-α, IL-4, IL-6, IL-10, IL-17, and IL-21 expression levels in the culture supernatant were measured by ELISA. Afterwards, CD4-PE and CD8-APC antibodies were added, and the proliferation of T cell subsets was detected by flow cytometry. Results A total of 10 MG patients were collected, including 3 males and 7 females with an average age of 19.25±6.28 years; and 15 control patients were collected, including 6 males and 9 females with an average age of 26.18±6.77 years. Compared with the control group, the mRNA and protein levels of α7 nAChR in the thymocytes of MG group were decreased, and the expression levels of IFN-γ, TNF-α, IL-4, IL-6 and IL-21 in the supernatant were increased (P<0.05), but there was no statistical difference in the expression of IL-10 and IL-17 (P>0.05). The cell-culture experiment showed that compared with the blank control group, the levels of IFN-γ, TNF-α, IL-6 and IL-21 secreted by T cells in the α7 nAChR antagonist group were increased (P<0.05), while they were decreased in the α7 nAChR agonist group (P<0.05). There was no statistical difference in the secretion levels of IL-4, IL-10 or IL-17 among the three groups (P>0.05). CD4+ T and CD8+ T cells in the α7 nAChR agonist group were significantly less than those in the blank control group and α7 nAChR antagonist group (P<0.001), while they were significantly more in the α7 nAChR antagonist group than those in the blank control group (P<0.001). Conclusion The expression of α7 nAChR in thymocytes of MG patients is decreased, and α7 nAChR may be involved in the inflammatory response in thymocytes and thus in thymic function.
