MiR-126 accelerates renal injury induced by UUO via inhibition PI3K/ IRS-1/ FAK signaling induced M2 polarization and endocytosis in macrophages.
Animals
MicroRNAs
/ genetics
Mice
Macrophages
/ metabolism
Signal Transduction
Phosphatidylinositol 3-Kinases
/ metabolism
RAW 264.7 Cells
Endocytosis
Insulin Receptor Substrate Proteins
/ metabolism
Ureteral Obstruction
/ metabolism
Kidney
/ metabolism
Focal Adhesion Kinase 1
/ metabolism
Male
Fibrosis
Disease Models, Animal
Mice, Inbred C57BL
Kidney Diseases
/ metabolism
M2 Macrophage Polarization
PI3K
Phagocytosis
UUO
miR-126
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
30 10 2024
30 10 2024
Historique:
received:
11
04
2024
accepted:
24
10
2024
medline:
31
10
2024
pubmed:
31
10
2024
entrez:
31
10
2024
Statut:
epublish
Résumé
To investigate the role and molecular mechanism of miR-126 in unilateral ureteral occlusion (UUO). We used bioinformatics to analyse miRNAs specifically expressed in UUO. The mouse model of UUO was established using RAW264.7 cells cultured in vitro and in vivo. The mice were divided into control group, miR-126-NC (negative control) group and miR-126-KD (knockdown) group. Then the relative expression of miR-126 was detected by quantitative reverse transcription-polymerase chain reaction (qRT-PCR), the renal fibrosis was detected by Masson staining, and the protein expression of CD68, collagen I and collagen III in the kidney was detected by immunofluorescence assay. Immunohistochemistry detects α-SMA expression. Moreover, Western blotting was performed to measure the expressions of p-PI3K, CD163, CD206, CD86, iNOS, IL-1β, p-FAK, p-Rac-1, p-IRS-1 and MMP9. The relative fluorescence intensity of F-actin and p-FAK was detected by immunofluorescence assay, and the phagocytosis ability of macrophages was determined by phagocytosis assay with fluorescent microspheres. Bioinformatics analysis reveals miR-126-specific overexpression in UUO. Successful transfection of miR-126-NC and miR-126-KD was confirmed by RT-PCR. The selective reduction of miR-126 was validated by Masson, immunohistochemistry and immunofluorescence staining to decrease the area of UUO-induced renal fibrosis and to lower the expression of CD68, α-SMA, collagen I, and collagen III. The reduction of iNOS expression may also be achieved with selective knockdown of miR-126, as verified by cell tests. enhances the phagocytic ability of macrophages and the expression of p-PI3K, CD206, p-FAK, F-actin, p-Rac-1, p-IRS-1 and MMP9. MiR-126 can inhibit the PI3K signaling pathway, promote M1 macrophage polarization, and suppress the activation of FAK and Rac-1, thus accelerating the progression of UUO.
Identifiants
pubmed: 39478171
doi: 10.1038/s41598-024-77691-1
pii: 10.1038/s41598-024-77691-1
doi:
Substances chimiques
MicroRNAs
0
Phosphatidylinositol 3-Kinases
EC 2.7.1.-
MIRN126 microRNA, mouse
0
Insulin Receptor Substrate Proteins
0
Focal Adhesion Kinase 1
EC 2.7.10.2
Ptk2 protein, mouse
EC 2.7.10.2
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
26083Subventions
Organisme : Cangzhou Science and Technology R&D Instruction Project
ID : No.183302086
Informations de copyright
© 2024. The Author(s).
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