Different Roles of M3 and α9 Acetylcholine Receptors in the Pathogenesis of Pemphigus Vulgaris: An In Vitro Experimental Study
Year: 2025
Type: article
Abstract: Objective: Pemphigus vulgaris (PV) is a severe autoimmune skin disease, and the role of acetylcholine receptor (AChR) in PV is still unclear. This study aimed to investigate the effects of M3 AChR and α9 AChR interference on acantholysis of keratinocytes, which may provide a deeper understanding of PV pathogenesis. Methods: This was an in vitro experimental study designed to explore the roles of M3 AChR and α9 AChR in PV pathogenesis. Sera from patients with PV and controls were collected from Hospital for Skin Diseases (Institute of Dermatology), Chinese Academy of Medical Sciences and Peking Union Medical College from January 1, 2016, to December 30, 2016, and were used to extract PV-immunoglobulin G (IgG). RNA interference was used to silence the M3 AChR gene or α9 AChR gene in HaCaT cells, after which the cells were treated with IgG from PV patients or controls. Changes in the adhesion of HaCaT cells were observed using a cell dissociation assay. Immunofluorescence assay was used to detect the staining patterns of desmosome proteins. Western blot analysis was performed to detect cytoskeleton- or noncytoskeleton-associated desmosome protein levels. The interactions of desmoglein-3 and plakoglobin were qualitatively detected using co-immunoprecipitation, and the p38 MAPK and epidermal growth factor receptor (EGFR) phosphorylation levels were examined. Two-tailed Student’s t -test was used to compare the data of the two groups. Results: With M3 AChR silencing, the HaCaT cells treated with PV-IgG-showed severe acantholysis compared to those with controls’ treatment ( P < 0.001); levels of cytoskeleton-associated desmoglein and plakoglobin were significantly reduced (both P < 0.001), whereas the phosphorylation of EGFR increased ( P = 0.001). In contrast, α9 AChR-silenced HaCaT cells exhibited fewer cell fragments than control HaCaT cells without α9 AChR silencing co-cultured with PV-IgG ( P < 0.001). The co-culture of α9 AChR-silenced cells with PV-IgG suppressed the internalization of desmoglein, and increased levels of cytoskeleton-associated desmoglein-3 and PG ( P < 0.001 and P = 0.002), and a close interaction between desmoglein-3 and plakoglobin was observed by immunoprecipitation ( P = 0.002). The phosphorylation levels of p38 MAPK and EGFR were suppressed by α9 AChR silencing (both P < 0.001). Conclusion: The findings of this study provided evidence that M3 AChR and α9 AChR may play roles in the pathogenesis of PV, demonstrating that M3 AChR exerts a protective role, whereas α9 AChR plays a pro-pathogenic effect in PV-IgG-induced acantholysis.
Authors: Zhi-Liang Li, Ling Yu, Yong-Hong Yang, Su-Ying Feng
Language: English
Subfield: Pathology and Forensic Medicine
Field: Medicine
Domain: Health Sciences
Open Access status: diamond
OpenAlex ID: W4417258102