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Calcium-activated chloride channel (CaCC) is an important ion channel of vascular smooth muscle cells (VSMCs). Recently, TMEM16A / Anoctamin-1 (ANO1) has been identified as the molecular basis of CaCC in VSMCs. Our previous study found that the overexpression of ANO1 in blood vessels of SHR rats might be involved in the pathophysiology of hypertension. In this study, we further observed the effect of ANO1 inhibitor T16A (inh) -A01(A01) on the proliferation and migration in primarily cultured VSMCs of SHR and its control Wistar-Kyoto (WKY) rats.
The cell proliferation was estimated by both MTT assay and Western blotting analysis of PCNA after the cells were treated by A01. The cell migration was evaluated by transwell test. The mRNA expressions of metalloproteinases-2 (MMP-2), metalloproteinases-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) of VSMCs were determined by real-time PCR.
Western Blotting indicated that the protein levels of both ANO1 and PCNA in SHR were significantly higher than that of WKY rats. In SHR rats, A01 reduced the cell viability and down-regulated the overexpression of PCNA. A01 also significantly suppressed the cell migration in SHR rats. Real-time PCR results showed that A01 decreased the mRNA expression of MMP2/9 but induced the TIMP-1 expression in SHR rats.
ANO1 inhibitor T16Ainh-A01 can inhibit the proliferation and migration of SHR VSMCs, but it has no obvious effect in WKY rats. This suggest that ANO1 inhibitor might be a promising candidate to prevent or reverse vascular remodeling in hypertension.