瑞舒伐他汀与阿托伐他汀对动脉钙化抑制作用的比较

Comparison of the inhibitory effects of rosuvastatin and atorvastatin on arterial calcification

目的 探讨瑞舒伐他汀和阿托伐他汀对动脉中膜钙化的抑制作用差异与其对骨保护素(OPG)/核因子κB受体活化因子配体(RANKL)比值的影响.方法 分别建立动脉中膜钙化的动物模型和细胞模型.6~8周龄体重180~200g雄性SD大鼠.使用华法林对大鼠灌胃诱导大鼠主动脉钙化构建动脉中膜钙化的动物模型,使用维生素C和β-磷酸甘油诱导大鼠主动脉中膜平滑肌细胞(SMC)钙化构建钙化的细胞模型.将80只大鼠采用简单随机表法分为4组:正常对照组,钙化组、瑞舒伐他汀组和阿托伐他汀组,每组20只.瑞舒伐他汀组和阿托伐他汀组在使用上述物质的同时分别再加入瑞舒伐他汀和阿托伐他汀进行干预;使用分光光度计测定钙离子含量,考马斯亮蓝法测定碱性磷酸酶(ALP)活性,蛋白质印迹法(Western blot)判断骨钙素水平,实时荧光定量聚合酶链反应(Real-time PCR)来检测此过程中OPG、RANKL的表达.多组间比较采用单因素方差分析ANOVA,组间的比较使用独立样本t检验.结果 动物模型中,瑞舒伐他汀组和阿托伐他汀组中的钙离子含量、ALP活性、骨钙素表达水平均低于钙化组[(0.94±0.09)mg/g比(1.44±0.10)mg/g、34.39±7.68 比 100.60±5.90、0.51±0.07 比 0.92±0.09,t=9.926、30.564、16.287,P<0.05;(1.30±0.29)mg/g 比(1.44±0.10)mg/g、61.47±4.04 比 100.60±5.90、0.74±0.06 比 0.92±0.09,t=2.092、24.463、7.528,P<0.05];瑞舒伐他汀组的钙离子含量、ALP活性、骨钙素表达水平均低于阿托伐他汀组[(0.94±0.09)mg/g 比(1.30±0.2)mg/g、34.39±7.68 比 61.47±4.04、0.51±0.07 比 0.74±0.06,t=5.152、13.946、10.887,P<0.05];瑞舒伐他汀组中动脉的 OPG/RANKL比值高于阿托伐他汀组(1.08±0.15比0.77±0.03,t=8.742,P<0.05).细胞模型中,瑞舒伐他汀组和阿托伐他汀组中的钙离子含量、ALP活性、骨钙素表达水平均低于钙化组[(69.83±3.77)μg/mg protein 比(133.00±4.50)μg/mgprotein、29.42±1.81 比99.94±3.83、0.49±0.06 比0.92±0.06,t=48.156、74.403、22.931,P<0.05;(99.03±2.93)μg/mg protein 比(133.00±4.50)μg/mgprotein、61.14±2.13 比 99.94±3.83、0.79±0.07 比 0.92±0.06,t=28.326、39.633、6.610,P<0.05];瑞舒伐他汀组的钙离子含量、ALP活性、骨钙素表达水平均低于阿托伐他汀组[(69.83±3.77)μg/mg protein 比(99.03±2.93)μg/mg protein、29.42±1.81 比 61.14±2.13、0.49±0.06 比 0.79±0.07,t=27.363、50.747、15.246,P<0.05];瑞舒伐他汀组中细胞的 OPG/RANKL 比值高于阿托伐他汀组(1.08±0.09比0.78±0.12,t=9.034,P<0.05).结论 瑞舒伐他汀比阿托伐他汀具有更明显的抑制钙化作用,其机制可能与其上调OPG/RANKL比值的能力更强有关.

Objective To explore the difference in inhibitory effects of rosuvastatin and atorvasta-tin on arterial media calcification and their effects on ratio of osteoprotectin(OPG)and receptor activator of nuclear factor-KB ligand(RANKL).Methods The animal model and cell model of arterial media calcifi-cation were established respectively.The 6-8 weeks old male SD rats,weighing 180-200 g,were purchased from Hunan Silaikejingda Experimental Animal Co.,Ltd.Arterial media calcification animal model was es-tablished by intragastric administration of warfarin in rats,and calcification cell model was established by calcification of rat aortic media smooth muscle cells(SMC)induced by vitamin C andβ-glycerol phos-phate.Totally,80 rats were randomly divided into 4 groups using a random number table method:normal control group,calcification group,rosuvastatin group,and atorvastatin group,with 20 rats in each group.Rosuvastatin group and atorvastatin group were treated with rosuvastatin and atorvastatin respectively.Calcium content was measured by spectrophotometer,alkaline phosphatase(ALP)activity was detected by Coomassie brilliant blue method,osteocalcin level was determined by Western blotting,and the expression of OPG and RANKL was detected by real-time quantitative polymerase chain reaction(Real-time PCR).One-way ANOVA was used for comparison between groups,and independent samples t test was used for comparison between two groups.Results In animal models,the calcium ion content,ALP activity,and osteocalcin expression levels in the rosuvastatin group and atorvastatin group were lower than those in the calcification group[(0.94±0.09)mg/g vs.(1.44±0.10)mg/g,34.39±7.68 vs.100.60±5.90,0.51±0.07 vs.0.92±0.09,t=9.926,30.564,16.287,P<0.05;(1.30±0.29)mg/g vs.(1.44±0.10)mg/g,61.47±4.04 vs.100.60±5.90,0.74±0.06 vs.0.92±0.09,t=2.092,24.463,7.528,P<0.05].Compared with the atorvastatin group,calcium ion content,ALP activity,and osteocal-cin expression levels were reduced in the rosuvastatin group[(0.94±0.09)mg/g vs.(1.30±0.2)mg/g,34.39±7.68 vs.61.47±4.04,0.51±0.07 vs.0.74±0.06,t=5.152,13.946,10.887,P<0.05].Compared with the atorvastatin group,the OPG/RANKL ratio of the arteries in the ro-suvastatin group was increased(1.08±0.15 vs.0.77±0.03,t=8.742,P<0.05).In the cell model,the calcium ion content,ALP activity,and osteocalcin expression levels in the rosuvastatin and atorvastatin groups were lower than those in the calcification group[(69.83±3.77)μg/mg protein vs.(133.00±4.50)μg/mg protein,29.42±1.81 vs.99.94±3.83,0.49±0.06 vs.0.92±0.06,t=48.156,74.403,22.931,P<0.05;(99.03±2.93)vs.(133.00±4.50)μg/mg protein,61.14±2.13 vs.99.94±3.83,0.79±0.07 vs.0.92±0.06,t=28.326,39.633,6.610,P<0.05].Compared with the atorvastatin group,calcium ion content,ALP activity,and osteocalcin expression levels were reduced in the rosuvastatin group[(69.83±3.77)μg/mg protein vs.(99.03±2.93)μg/mg protein,29.42±1.81 vs.61.14±2.13,0.49±0.06 vs.0.79±0.07,t=27.363,50.747,15.246,P<0.05].Com-pared with the atorvastatin group,the OPG/RANKL ratio of cells in the rosuvastatin group was increased(1.08±0.09 vs.0.78±0.12,t=9.034,P<0.05).Conclusion Rosuvastatin has more obvious anti-calcification effect than atorvastatin,and its mechanism may be related to its stronger ability to upregulate the OPG/RANKL ratio.