β-磷酸甘油诱导体外培养大鼠主动脉平滑肌细胞的钙化

β-glycerophosphate induced calcification of rat aortic vascular smooth muscle cells

目的:研究β磷酸甘油对体外培养的大鼠主动脉平滑肌细胞的钙化作用。方法:采用组织块培养法体外培养大鼠主动脉平滑肌细胞。细胞分为钙化组和正常组。钙化组加入10mmol/Lβ磷酸甘油,0.1μmol/L胰岛素及50μg/L维生素C(钙化培养基)诱导细胞钙化,继续培养14d。茜素红S染色观察钙结节计数并测定细胞钙沉积含量。采用四唑盐比色法(MTT)测量细胞增殖。结果:钙化组钙结节计数为(7.4±3.8)%,较正常组(4.3±1.9)%显著增加(P<0.05);细胞钙沉积含量为(5.4±0.4)mmol/g蛋白,较正常组(1.2±0.5)mmol/g蛋白显著增加(P<0.05)。MTT检测细胞增殖,钙化组细胞增殖为0.071±0.011,较正常组0.040±0.009明显增加(P<0.01)。结论:β磷酸甘油能诱导体外培养的大鼠主动脉平滑肌细胞的钙化,钙化能促进平滑肌细胞的增殖。

AIM: To study calcification of rat aortic vascular smooth muscle cells (VMSC) induced by β-glycerophosphate in vitro. METHODS: VMSC was cultured in vitro by tissue explant. All cells were divided into calcification group and normal group. In calcification group, 10 mmol/L β-glycerophosphate, 0.1 μmol/L insulin and 50 μg/L Vitamin C were added to induce VSMC calcification. All groups were continuously treated for 14 days. Alizarin red S staining was applied to count the calcium nodus, and cell calcium depositions were measured. VSMC proliferation was measured by tetrazolium salt (MTT) methord. RESULTS: The count of calcium nodus in calcification group was (7.4±3.8)%, which were significantly increased compared to normal group(4.3±1.9)%, (P<0.01). Cell calcium depositions in calcification group were (5.4±0.4) mmol/g protein, compared to normal group (1.2±0.5) mol/g protein, were significantly increased, and had reached a statistics difference (P<0.01). VSMC proliferation in calcification group was (0.071±0.011, compared to normal group (0.040±0.009, was significantly increased, and had reached statistics difference (P<0.01). CONCLUSION: β-glycerophosphate can induced calcification of rat aortic VSMC in vitro. Calcification can promote VSMC proliferation.