Renin Increases Calcification of Rat Vascular Smooth Muscle Cells Through Angiotesin Ⅱ-independent Mechanisms
|Course||Department of Cardiology|
|Keywords||Vascular calcification Renin Vascular smooth muscle cells AngiotensinⅡ|
Objective:To explore the impact of renin through angiotesin Ⅱ-independent mechanisms on calcification of rat vascular smooth muscle cells in vitro, and its molecular mechanisms.Methods:Primary cultured cells were obtained by tissue-piece inoculation. Calcification of cultured rat vascular smooth muscle cells was produced by incubation with (3-glycerophosphate and sodium pyruvate. The vascular smooth muscle cells cells were divided into6groups:the normal group (cultured in normal medium), the calcification group (incubated in calcified medium), the calcification+angiotesin Ⅱ receptor blocker group(cultured in calcified medium, giving Losartan10-6mol/L to block AT1and PD123,31910-5mol/L to block AT2),calcification+renin group (incubated in calcified medium, giving Losartan10-6mol/L and PD123,31910-5mol/L and10-10,10-9,10-8mmol/L renin). Calcification was confirmed by Von Kossa staining. Calcium content and alkaline phosphatases (ALP) activity were measured to estimate the extent of Calcification. The mRNA expression of core binding factor1(Cbfα1) and the transforming growth factor (31(TGFβ1) was measured by competitive quantitative RT-PCR.The expression of Cbfal protein content was measured by western blotting.Results:Compared with the blank control group, the calcium content and ALP activity of the calcification group increased significantly, as well as the expression of mRNA of Cbfα1,TGFβ1and Cbfal protein expression increased significantly (P <0.01). Compared with calcification group, calcification+angiotensin Ⅱ receptor blocker group had no difference on calcification of vascular smooth muscle cell. Compared with calcification+angiotensin Ⅱ receptor blocker group, intervention of the renin (10-10、10-9、10-8mmol/L) in a dose-dependent manner to further promote the calcium content and the ALP activity of rat vascular smooth muscle cells, as well as the mRNA expression of Cbfα1,TGFβ1and Cbfal protein expression (P<0.05).Conclusions:Renin can promote β-glycerophosphate-induced calcification of vascular smooth muscle cells through angiotesin Ⅱ-independent mechanisms, the mechanisms of renin promote calcification of vascular smooth muscle cells through angiotesin Ⅱ-independent way may be though promoting the expression of TGFβ1、Cbfα1.