Effects of Notch signaling pathway on the adipogenic differentiation of infantile hemangioma-derived mesenchymal stem cells
10.3760/cma.j.cn114453-20220322-00082
- VernacularTitle:Notch信号通路对婴幼儿血管瘤间充质干细胞成脂分化的影响
- Author:
Weidong WANG
1
;
Weimin SHEN
;
Tao HAN
;
Yuan WANG
;
Sheng CHEN
;
Jiawei LIN
Author Information
1. 南京医科大学附属儿童医院烧伤整形科，南京 210008
- Keywords:
Hemangioma;
Mesenchymal stem cells;
Notch signaling pathway;
Adipogenic differentiation
- From:
Chinese Journal of Plastic Surgery
2023;39(7):762-769
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To study the effects and potential mechanisms of Notch signaling pathway on the adipogenic differentiation of infantile hemangioma-derived mesenchymal stem cells.Methods:From January 2021 to June 2021, 10 infantile hemangioma specimens (6 males and 4 females, aged 2 months to 6 months, average age 3.5 months) were collected from the Department of Burns and Plastic Surgery of the Children’s Hospital of Nanjing Medical University. Hem-MSCs were isolated from hemangioma in the proliferating phase by adherent screening and identified by flow cytometry. Adipogenic induction was performed after successful identification. Real-time quantitative fluorescence PCR was used to detect the expression of Notch1, Jagged1 and Hes1 genes after 14 days of adipogenesis induction of Hem-MSCs. The Notch signaling pathway inhibitor DAPT, PI3K/Akt signaling pathway agonist 740Y-P and solvent DMSO were added to Hem-MSCs, and adipogenesis induction was performed. The cells were divided into six groups: control group, adipogenic induction group, adipogenic induction+ DMSO group, adipogenic induction+ DAPT group, adipogenic induction+ 740Y-P group, and adipogenic induction+ 740Y-P+ DAPT group. After 14 days of adipogenic induction, real-time quantitative fluorescence PCR was used to detect the expression of PPARγ and C/EBPα genes in the key transcription factors of adipogenic differentiation in the above six groups. Oil red O staining was applied to identify the adipogenic differentiation capabilities; after adipogenic induction for 7 days, the expression levels of p-PI3K, PI3K, p-AKT, and AKT proteins in the adipogenic induction+ DMSO group, adipogenic induction+ DAPT+ 740Y-P group were detected by western blotting. In comparing the two groups, an unpaired Student’s t-test was used to assess the differences, and analysis of variance was applied for the analysis of the mean values among multiple groups. and P<0.05 was considered statistically significant. Results:The expression levels of Notch1, Jagged1 and Hes1 decreased gradually during the adipogenic induction of Hem-MSCs, and the differences were statistically significant ( tNotch1=8.99, PNotch1=0.008; tJagged1=9.49, PJagged1=0.007; tHes1=7.74, PHes1=0.015). After the addition of Notch signaling pathway inhibitor DAPT during the adipogenic induction of Hem-MSCs, the expression of proteins associated with PI3K/AKT signaling pathway decreased, and the difference was statistically significant ( qp-PI3K=4.78, Pp-PI3K=0.014; qp-AKT=5.04, Pp-AKT=0.010) and the expression of adipogenesis indexes increased( qOil red O=6.07, POil red O=0.003; qPPARγ=17.34, PPPARγ<0.001; qC/EBPα=14.8, PC/EBPα<0.001). There was no significant change in adipogenesis indexes after addition of PI3K/Akt signaling pathway agonist 740Y-P ( qOil red O=1.82, POil red O=0.786; qPPARγ=0.97, PPPARγ=0.981; qC/EBPα=1.98, PC/EBPα=0.654). The expression of adipogenesis indexes decreased after adding DAPT and 740Y-P( qOil red O=5.22, POil red O=0.013; qPPARγ=9.78, PPPARγ<0.001; qC/EBPα=16.74, PC/EBPα<0.001). Conclusion:The inhibition of Notch signaling pathway can promote the adipogenic differentiation of Hem-MSCs by reducing the activity of PI3K/Akt signaling pathway.
