Background
MiR-10a-3p is associated with the pathogenesis of many immune inflammatory diseases including Mycoplasma pneumoniae pneumonia (MPP), and cytochrome coxidase assembly homologue 11 (COX11) is one of its direct target proteins. This study investigates the function and mechanism of miR-10a-3p targeting with COX11 in the development and progression of paediatric MPP.
Conclusions
In paediatric MPP, increased miR-10a-3p downregulated COX11, activating NF-κB signalling pathway to promote disease development and progression.
Methods
Ninty-seven paediatric MPP patients and 100 age- and sex-matched healthy children were enrolled. Clinical and laboratory indicators of paediatric MPP patients were collected. The mRNA levels of the COX11 gene and miR-10a-3p were detected by qRT-PCR. THP-1 mononuclear macrophages were stimulated using MPP lipid-associated membrane proteins (Mp-LAMPs). The relative expression level of miR-10a-3p was detected after 12, 24, and 48 h. THP-1 cells were transfected to overexpress or inhibit the expression of miR-10a-3p, miR-10a-3p, COX11 mRNA, NF-κB signalling pathway-related proteins, and C-reactive protein (CRP) were detected after 48 h by Western blot.
Results
The relative expression level of miR-10a-3p in the MPP group was 2.38±0.52, compared with 1.76±0.38 in control group (t=4.584, P<0.001) whileCOX11 in MPP group was 3.70±1.12, compared to 5.78±1.84 in control group (t=4.876, P<0.001). Pearson correlation analysis showed that miR-10a-3p and COX11 in MPP group presented a negative correlation (r=-0.679, P<0.001). By searching in the prediction website of TargetScan database, it was found that miR-10a-3p and Cox11 genes had targeted regulatory binding sites, and the targeting relationship between miR-10a-3p and Cox11 genes was confirmed by dual luciferase reporting assay in 293T cells. Among paediatric MPP patients, miR-10a-3p expression had a positive correlation with the white blood cells count, erythrocyte sedimentation rate (ESR), and CRP expression, while COX11 mRNA expression had a positive correlation with ESR and CRP. After LAMP stimulation, the miR-10a-3p expression level in THP-1 cells significantly increased (P<0.05). After THP-1 cells were transfected with the miR-10a-3p mimic or inhibitor, the relative expression level of miR-10a-3p significantly increased or decreased, respectively. COX11 expression in the mimic group significantly decreased, whereas COX11 in the inhibitor group significantly increased (both P<0.05). In addition, after transfection, IκBα expression significantly decreased and that of p-IKKα/β, p-p65, and CRP significantly increased in the mimic group, and the opposite was true in the inhibitor group. Conclusions: In paediatric MPP, increased miR-10a-3p downregulated COX11, activating NF-κB signalling pathway to promote disease development and progression.