Effects of C-C motif chemokine receptor 2 on phenotypic transformation of cardiac fibroblasts after hypoxia / 解放军医学杂志

ABSTRACT

Objective To investigate the effect of C-C motif chemokine receptor 2 (CCR2) on phenotypic transformation of cardiac fibroblasts after hypoxia. Methods The mouse myocardium primary fibroblasts were extracted by collagenase digestion. Cells were divided into control, hypoxia-24h and hypoxia-48h, the mRNA and protein expression of CCR2 was examined by real-time PCR and Western blotting. CCR2 low expression cell (si-CCR2) was established using by small interfering RNA. Cells were divided into four groups including si-control, si-CCR2, si-control+hypoxia, si-CCR2+hypoxia. The mRNA and protein expressions of CCR2, α smooth muscle actin (α-SMA) and Collagen 1A (Col 1A) were detected by real-time PCR and Western blotting, cell proliferation was detected by Cell Counting Kit-8 (CCK8) and Bromodeoxyuridine (BrdU). Results Compared with control, the mRNA and protein levels of CCR2 significantly increased in hypoxia-24h and hypoxia-48h group (P<0.01), however, no significance was found in these two time points. Compared with si-control group, the mRNA and protein expressions of CCR2, α-SMA and Col 1A not significantly changed in si-CCR2 group. Compared with si-control group, the mRNA and protein levels of CCR2, α-SMA and Col 1A significantly increased in si-control+hypoxia group (P<0.01 or P<0.05), and cell proliferation increased (P<0.01). Compared with si-control+hypoxia group, the protein expression of α-SMA and Col 1A decreased in si-CCR2+hypoxia group (P<0.05), and cell proliferation also decreased in si-CCR2+hypoxia group (P<0.05). Conclusions CCR2 could affect the phenotypic transformation of cardiac fibroblasts after hypoxia.