Regulatory effect of FGF2 on the expression of R-spondin 1 in mouse intestinal stromal cells
10.3969/j.issn.1674-8115.2025.08.001
- VernacularTitle:FGF2对小鼠肠道间质细胞R-spondin 1表达的调控作用
- Author:
Jingcong LI
1
;
Han ZHAO
;
Qiaowen LIN
;
Hongxiang SUN
;
Bing SU
;
Ningbo WU
Author Information
1. 上海交通大学基础医学院免疫学与微生物学系,上海 200025;上海交通大学医学院上海市免疫学研究所,上海 200025
- Publication Type:Journal Article
- Keywords:
R-spondin 1 gene;
fibroblast growth factor 2(FGF2);
CD34+CD81+stromal cell;
mitogen-activated protein kinase(MAPK)pathway
- From:
Journal of Shanghai Jiaotong University(Medical Science)
2025;45(8):939-948
- CountryChina
- Language:Chinese
-
Abstract:
Objective·To preliminarily investigate the regulatory effect and underlying mechanism of fibroblast growth factor 2(FGF2)on R-spondin 1(Rspo1)expression in CD34+CD81+stromal cells from the mouse colon.Methods·Colonic CD45-CD326-CD31-GP38+CD81+Rspo1-tdTomato+stromal cells were sorted from Rspo1-tdTomato reporter mice by flow cytometry and subsequently cultured in vitro.The expression of surface protein markers was evaluated by flow cytometry after 14 d of culture.qPCR was employed to quantify Rspo1 expression in response to stimulation with FGF2,FGF9,epidermal growth factor(EGF),platelet-derived growth factor-bb(PDGF-bb),insulin-like growth factor 1(IGF1),or hepatocyte growth factor(HGF).RNA sequencing and bioinformatic analyses were used to identify the signaling pathways underlying FGF2-mediated regulation of Rspo1,followed by preliminary validation with pathway-specific inhibitors and qPCR.Results·After 14 d of culture,the sorted colonic stromal cells retained expression of CD34,CD81,and glycoprotein GP38,while remaining negative for other lineages markers CD45,CD326,and CD31.qPCR revealed that 20 ng/mL FGF2 significantly suppressed Rspo1 expression,whereas the other tested growth factors exerted no notable effect.RNA sequencing and bioinformatic analysis indicated that mitogen-activated protein kinase(MAPK)signaling pathway played a key role in the regulatory effect of FGF2 on Rspo1.qPCR further demonstrated that pretreatment with U0126,an inhibitor of mitogen extracellular kinase 1/2(MEK1/2),reversed FGF2-mediated suppression of Rspo1 expression.Conclusion·FGF2 may inhibit Rspo1 expression in mouse colonic CD34+CD81+stromal cells via the MEK1/2-extracellular regulated protein kinase 1/2(ERK1/2)signaling pathway.
