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目的:研究华法令对大鼠体内主动脉的钙化作用。方法:采用华法令(3 mg/g饲料)的饲料喂养诱导大鼠体内动脉钙化。实验分3组(n=6),对照组、6 W钙化组、12 W钙化组。Von Kossa染色观察钙结节形成,邻甲酚酞络合酮比色法定量测定钙沉积含量。采用TUNEL染色法检测大鼠主动脉组织内血管平滑肌细胞(Vascular Smooth Muscle Cell,VSMC)凋亡。Western blot法检测大鼠主动脉组织中生长抑制特异蛋白6(Growth Arrest Specific Gene 6 Protein,Gas 6)蛋白表达水平。结果:钙化组钙结节、钙沉积含量及VSMC凋亡均高于对照组(P<0.01)有显著差异。钙化形成与凋亡表达呈显著正相关(R2=0.8853,P<0.0001);钙化组Gas 6蛋白表达量较对照组下调(P<0.01)有显著差异。结论:华法令通过下调Gas 6蛋白诱导大鼠体内主动脉钙化形成。
Objective: To study the effects of warfarin on calcification of aorta in rats. Methods: Arterial calcification was induced in rats by using warfarin (3 mg / g diet). The experiment was divided into 3 groups (n = 6), control group, 6 W calcification group, 12 W calcification group. The formation of calcium nodules was observed by Von Kossa staining and the calcium deposition was quantitatively determined by o-cresolphthalein complex ketone colorimetry. TUNEL staining was used to detect the apoptosis of Vascular Smooth Muscle Cell (VSMC) in aorta of rats. Western blot was used to detect the protein expression of growth arrest protein 6 (ARF6) in rat aortic tissue. Results: Calcification, calcium deposition and VSMC apoptosis in the calcified group were significantly higher than those in the control group (P <0.01). There was a significant positive correlation between calcification and apoptosis (R2 = 0.8853, P <0.0001). The expression of Gas 6 in calcification group was significantly lower than that in control group (P <0.01). Conclusion: Warfarin can induce the formation of aortic calcification in rats by down-regulating Gas 6 protein.