Abstract 060: Role of Mir-214 in the Regulation of Perivascular Fibrosis in Angiotensin II Induced Hypertension

Abstract

Objective: Hypertension (HT) is associated with perivascular inflammation and increased vascular fibrosis. MicroRNAs (miR) are a novel gene expression regulation mechanism and play a pivotal role in a range of pathological processes. The role and mechanism of miR214 in vascular fibrosis is unknown. Methods: 3-month-old C57BL/6, miR214KO and wild-type littermates were treated with angiotensin II (AngII, 490ng/kg/min; n=6-10) or control buffer for 14 days. PVATs from C57BL/6 animals were analysed using TaqMan_Rodent_microRNA_Arrays. Histological studies, wire myography, lucigenin-enhanced luminometry and cytometrical analysis was conducted, followed by statistical analysis with ANOVA or t-test. Data are expressed as a mean±SEM. Results: Out of 381 miRs, 16 were significantly overexpressed in C57BL/6 AngII animals, with only miR214 showing 8-fold induction (p<0.01) after Bonferroni correction. Also, 3-fold elevation of pri-miR-214 was observed. Interestingly, hydralazine treatment prevented both these changes (p<0.01). AngII infusion in miR214 KO animals did not alter blood pressure when compared to WT mice. Mir214 KOs exhibited diminished peri-aortic fibrosis (44779±2491 vs 78805±8696μm, p<0.01), upon AngII hypertension. This was associated with a significantly reduced induction of COL1A1, COL3A1 and TGFβ1 mRNA expression in PVAT and aortas (p<0.05). Vascular studies revealed improved endothelial function (69±10 vs. 22±4%, p<0.01), protection against oxidative stress (66±7 vs 118±19 RLU/sec/mg, p<0.001) and NOX2 mRNA expression (1.9±0.2 vs1.1±0.1, p<0.05) in AngII miR-214-KO aortas, while these parameters were not altered in mesenteric arteries. Recruitment of T cells into aortic PVAT was abolished in KO HT animals in comparison to control group (192±65 vs. 603±164 cell/mg; p<0.05). AngII HT was associated with 4-fold increase of miR-214 expression in the circulating peripheral blood T cells and 2-fold in the spleen. Moreover, AngII infusion increased TNFα mRNA expression in WT T cells (1±0.1 vs 1.6±0, p<0.01) whereas this effect was not seen in miR214 KO T cells (0.9±0.3 vs 0.9±0.1). Conclusions: MiR-214 plays a major role in modulation of aortic fibrosis, vascular function, oxidative stress and perivascular inflammation.