Macrophage Phenotype Induced by Circulating Small Extracellular Vesicles from Women with Endometriosis
Abstract
:1. Introduction
2. Methods
2.1. Patients
2.2. Sample Collection
2.3. sEV Isolation
2.4. Nanoparticle Tracking Analysis
2.5. SDS-PAGE and Western Blot
2.6. Cell Culture
2.7. RNA Isolation and qPCR
2.8. sEVs and Cells Staining
2.9. Confocal Microscopy
2.10. Statistical Analysis
3. Results
3.1. Patient Characteristics
3.2. sEVs Characterization
3.3. Macrophage Exosome Uptake
3.4. Changes in Macrophage Gene Expression and Phenotype
3.5. Effect of PPARG Activation
4. Discussion
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Gene | Accession Number | Product Size | Sequence |
---|---|---|---|
GAPDH | NM 002046.3 | 97 bp | Forward: 5′-GATCATCAGCAATGCCTCCT-3′ Reverse: 5′-TGTGGTCATGAGTCCTTCCA-3′ |
IL1B | NM 000576.2 | 120 bp | Forward: 5′-GGACAAGCTGAGGAAGATGC-3′ Reverse: 5′-TCGTTATCCCATGTGTCGAA-3′ |
TNF | NM 000594.4 | 114 bp | Forward: 5′-AGCCCATGTTGTAGCAAACC-3′ Reverse: 5′-GGCACCACCAACTGGTTATC-3′ |
MRC1 | NM 002438 | 131 bp | Forward: 5′-GGATGGATGGCTCTGGTG-3′ Reverse: 5′-TCTGGTAGGAAACGCTGGT-3′ |
ARG1 | NM 000045.3 | 160 bp | Forward: 5′-ACACTCCATTGACAACCACA-3′ Reverse: 5′-TCCACGTCTCTCAAGCCAAT-3′ |
PPARG | NM 138712.3 | 208 bp | Forward: 5′-TTGCAGTGGGGATGTCTCAT-3′ Reverse: 5′-TTTCCTGTCAAGATCGCCCT-3′ |
CD36 | NM 001127444.1 | 150 bp | Forward: 5′-AGATGCAGCCTCATTTCCAC-3′ Reverse: 5′-GCCTTGGATGGAAGAACAAA-3′ |
Control Group (n = 12) | Endometriosis Group (n = 12) | p Value | |
---|---|---|---|
Age (years) | 33.1 ± 3.6 | 34.2 ± 4.1 | 0.14 |
BMI (Kg/m2) | 24.5 ± 4.6 | 24.8 ± 3.9 | 0.56 |
Tobacco use | 3 (25) | 4 (33.3) | 0.21 |
Nulliparous | 5 (41.6) | 6 (50) | 0.30 |
Dysmenorrhea (NRS) | 0.7 ± 2.3 | 8.7 ± 3.2 | <0.001 |
Chronic pelvic pain (NRS) | 0.0 ± 0.0 | 7.4 ± 2.3 | <0.001 |
Dyspareunia (NRS) | 0.0 ± 0.0 | 6.5 ± 2.4 | <0.001 |
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Martínez-Zamora, M.A.; Armengol-Badia, O.; Quintas-Marquès, L.; Carmona, F.; Closa, D. Macrophage Phenotype Induced by Circulating Small Extracellular Vesicles from Women with Endometriosis. Biomolecules 2024, 14, 737. https://doi.org/10.3390/biom14070737
Martínez-Zamora MA, Armengol-Badia O, Quintas-Marquès L, Carmona F, Closa D. Macrophage Phenotype Induced by Circulating Small Extracellular Vesicles from Women with Endometriosis. Biomolecules. 2024; 14(7):737. https://doi.org/10.3390/biom14070737
Chicago/Turabian StyleMartínez-Zamora, María Angeles, Olga Armengol-Badia, Lara Quintas-Marquès, Francisco Carmona, and Daniel Closa. 2024. "Macrophage Phenotype Induced by Circulating Small Extracellular Vesicles from Women with Endometriosis" Biomolecules 14, no. 7: 737. https://doi.org/10.3390/biom14070737
APA StyleMartínez-Zamora, M. A., Armengol-Badia, O., Quintas-Marquès, L., Carmona, F., & Closa, D. (2024). Macrophage Phenotype Induced by Circulating Small Extracellular Vesicles from Women with Endometriosis. Biomolecules, 14(7), 737. https://doi.org/10.3390/biom14070737