中国病理生理杂志2025，Vol.41Issue(4)：825-832,8.DOI:10.3969/j.issn.1000-4718.2025.04.025
基于小鼠P815与N2a细胞体外共培养构建模拟肠易激综合征内脏高敏神经增生细胞模型
Established cell model and mechanism of visceral hypersensitivity and nerve hyperplasia in IBS using P815 and N2a co-culture
摘要
Abstract
AIM:To establish a cell model of visceral hypersensitivity and nerve hyperplasia in irritable bowel syndrome(IBS)by conducting an in vitro co-culture of mouse P815 mast cells and N2a nerve cells and explore its possible mechanism.METHODS:Enzyme-linked immunosorbent assay(ELISA)with three replicates was used to confirm the C48/80-induced P815 degranulation.The length of neurites was observed under bright field microscopy to determine the number of differentiated neurons,thereby selecting the concentration of retinoic acid(RA)for stimulating the differentia-tion of N2a cells,with four replicates.A co-culture system of P815 and N2a cells was established using Transwell cham-bers with four replicates.The following groups were established:N2a cells cultured alone,N2a cells co-cultured with P815 cells,N2a cells co-cultured with P815 cells plus C48/80,and N2a cells plus RA group.After co-culturing,the num-ber of differentiated N2a cells was observed under bright field.The expression of nerve growth factor(NGF),tyrosine ki-nase receptor A(TRKA),growth-associated protein-43(GAP43),neuron-specific enolase(NSE),synapsin(SYN),and postsynaptic density protein-95(PSD-95)at both protein and gene levels in N2a cells was detected using Western blot and polymerase chain reaction(PCR),with four replicates.RESULTS:The best condition for N2a differentiation was stimulation with 10 µmol/L RA for 24 hours,whereas the best condition for degranulation was stimulation of P815 cells with 20 mg/L C48/80 for 24 hours.Compared with N2a cells cultured alone,the differentiation ratio of the N2a+P815+C48/80 and N2a+RA groups was significantly increased(P<0.01),and the protein and mRNA expressions of NGF,TRKA,GAP43,NSE,SYN,and PSD-95 were significantly increased(P<0.05).CONCLUSION:Our results revealed that mast cell degranulation enhances the level of nerve hyperplasia in enteric nerve cells and promotes changes in nerve structure and function.Synaptic remodeling regulated by abnormal expression of key proteins such as NGF,TRKA,and GAP43 is involved in the nerve hyperplasia induced by mast cell degranulation.关键词
肠易激综合征/内脏高敏/肠神经系统/神经可塑性Key words
irritable bowel syndrome/visceral hypersensitivity/enteric nervous system/neural plasticity分类
医药卫生引用本文复制引用
李鸿斌,甘春莉,谢翔雨,刘珊,卢琴,柯威,齐诗语,黄育生,唐洪梅..基于小鼠P815与N2a细胞体外共培养构建模拟肠易激综合征内脏高敏神经增生细胞模型[J].中国病理生理杂志,2025,41(4):825-832,8.基金项目
广东省基础与应用基础研究基金项目(No.2023A1515220155) （No.2023A1515220155）
国家中医药传承创新中心科研专项(No.2023ZJ03) （No.2023ZJ03）
广东省医院协会药学科研专项(No.2021YXZD02) （No.2021YXZD02）
广东省普通高校重点领域专项(No.2023ZDZX2019) （No.2023ZDZX2019）
