HMEC-EXOS-derived hsa-miR-4488_L regulates osteogenic differentiation by targeting Smad3
10.3760/cma.j.issn.0254-9026.2025.07.013
- VernacularTitle:血管内皮细胞外泌体来源的hsa-miR-4488_L通过靶向 Smad3缓解衰老相关骨丢失
- Author:
Yun CHEN
1
;
Wenjie CHEN
;
Yajun CHEN
;
Jieyu HE
;
Junkun ZHAN
;
Qiong LU
Author Information
1. 中南大学湘雅二医院药学部，长沙 410011
- Publication Type:Journal Article
- Keywords:
Osteoporosis;
hsa-miR-4488_L;
BMSCs;
Osteogenic differentiation;
Adipogenic differentiation
- From:
Chinese Journal of Geriatrics
2025;44(7):925-932
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the role of human dermal microvascular endothelial cells(HMEC-1)exosome hsa-miR-4488_L in regulating the osteogenic and lipogenic differentiation of bone marrow mesenchymal stem cells(BMSCs)and to elucidate the mechanism of action underlying fate differentiation.Methods:The hsa-miR-4488_L mimic or negative control mimic was transfected into BMSCs, which were then cultured under osteogenic or lipogenic conditions, respectively.RT-qPCR was employed to detect the mRNA expression levels of osteogenic and lipogenic genes in BMSCs.Alizarin red staining was utilized to assess the osteogenic differentiation capability of hsa-miR-4488_L in BMSCs, while oil red O staining was used to evaluate the lipogenic differentiation potential of BMSCs.The mimic control and hsa-miR-4488_L mimic were transfected into elderly BMSCs, and age-related phenotypes were assessed using RT-qPCR and SA-β-gal staining.The direct target genes of hsa-miR-4488_L acting on BMSCs were identified through bioinformatics analysis and subsequently validated by RT-qPCR and Western blot.Results:After treatment with the hsa-miR-4488_L mimic, the expressions of osteogenic-related genes ALP( P=0.007), BSP( P=0.001), and Col1( P<0.001)in BMSCs were upregulated.Additionally, alizarin red staining results indicated an increase in the number of calcified nodules Pparγ( P=0.002).Concurrently, under adipogenic induction conditions, the adipogenic-related genes Pparγ( P=0.008)and Perilipin( P<0.001)were significantly downregulated in the hsa-miR-4488_L mimic treatment group, and oil red O staining demonstrated a reduction in lipid droplet production( P=0.032).The mRNA expression of the aging-related gene P16( P=0.009)was downregulated following treatment with the hsa-miR-4488_L mimic, and the number of senescent cells decreased as indicated by SA-β-gal staining.Bioinformatics analysis revealed that Smad3 was the direct target gene of hsa-miR-4488_L in BMSCs.RT-qPCR results confirmed that the expression of Smad3 was downregulated after treatment with the hsa-miR-4488_L mimic( P=0.040).Furthermore, Western blot analysis showed a reduction in the protein level of Smad3. Conclusions:HMEC-EXOs-derived hsa-miR-4488_L regulates the balance between osteogenic and adipogenic differentiation in BMSCs through Smad3.Consequently, hsa-miR-4488_L may serve as a potential miRNA biomarker for age-related osteoporosis.
