Calcification of aortic smooth muscle cells from high-sugar and high-fat diet rat cultured in vitro

Objective To investigate the mechanisms of diabetic vascular calcification by examining the markers related to vascular calcification of aortic smooth muscle cells (SMCs) isolated from the thoracic aortas of Goto-Kakizaki(GK) rats fed with high-sugar and high-fat diet.Methods GK and Wistar rats were fed with high-sugar and high-fat diet for two weeks.SMCs were isolated from GK rats and Wistar rats simultaneously,and SMCs from Wistar rats were used as control.The growth of SMCs was observed by cell counting.The calcium contents in SMCs layer and supernatant were measured by methyl thymol blue method.Expression of alkaline phosphatase (ALP),osteopontin (OPN),core binding factor α-1 (Cbfα-1),α-smooth muscle actin (α-SMA) were detected by real-time quantitative PCR.Results Compared with SMCs from Wistar rats,SMCs from GK rats growed slowly significantly (F =363.392,P ＜0.05),the calcium contents was significantly increased in the cells' layer [(0.56 ± 0.22) vs.(0.39 ± 0.09),t =2.47,P ＜0.05],and was significantly reduced in the supernatant [(0.82 ± 0.22) vs.(1.20 ± 0.17),t =-22.573,P ＜0.05] in GK rats group.The expression of ALP (t=12.963,P ＜0.05),OPN (t=8.305,P ＜0.05),Cbfα-1 (t =10.109,P ＜0.05) in SMCs from GK rats increased,while expression of α-SMA decreased (t =-8.219,P ＜0.05).Conclusions SMCs from high-sugar and high-fat fed GK diabetic rats tend to appear calcification. Key words: Cell calcification;  Aortic smooth muscle cells;  Goto-Kakizaki rat;  Diabetes mellitus