维生素D_3联合尼古丁诱导的血管钙化亚慢性模型的建立

目的建立维生素D3(vitamin D3)联合尼古丁(nico-tine)诱导的大鼠血管钙化亚慢性动物模型。方法采用维生素D3(300 kIU.kg-1)肌注和尼古丁(25 mg.kg-1,5 ml.kg-1)灌胃制备大鼠血管钙化亚慢性模型,定期测定大鼠体重和尾动脉血压;在造模的第8周、第16周,分别测定大鼠心脏系数、血浆钙、磷含量、血管钙含量、血管碱性磷酸酶(alkaline phosphatase,ALP)活性和血管钙沉积等与血管钙化相关的指标,对胸主动脉进行Von Kossa染色。结果第3~8周,VDN(vitamin D3and nicotine)大鼠尾动脉压持续增加,与对照组相比差异均有显著性(P均<0.05)。第9周后,仍然维持在较高水平。与对照组相比,VDN组大鼠第8周心脏系数升高8.4%(P<0.01);血钙、血磷分别升高3.2%(P<0.01)和12.0%(P<0.05);血管组织钙含量升高27.0%(P<0.01);血管ALP活性与血管钙沉积分别高151.8%和68.8%(P均<0.01)。与对照组相比,第16周VDN组大鼠心脏系数升高7.7%(P<0.01),血钙升高3.2%(P<0.05),血磷降低1.4%;血管组织钙含量升高51.0%(P<0.01);血管ALP活性与血管钙沉积分别高87.2%(P<0.01),31.6%(P<0.05)。Von Kossa染色结果显示第8周和第16周VDN组大鼠胸主动脉中膜均有大量深色颗粒沉积。结论维生素D3和尼古丁诱导的16 wk大鼠血管钙化亚慢性模型可作为药物筛选和机制研究的疾病模型。

Establishment of a sub-chronic model of vascular calcification induced by hypervitaminosis D3 united with nicotine

Aim To establish a rat aortic calcification sub-chronic model induced by vitamin D3 and nicotine.Methods The vascular calcification model was produced by vitamin D3(300 kIU·kg-1,im) plus nicotine(25 mg·kg-1,5 ml·kg-1,po).Rat body weight and blood pressure were monitored on a regular basis.Ratio of heart to body weight,plasma calcium,plasma phosphate content,vascular calcium content,Von Kossa staining,activity of alkaline phosphatase(ALP) and 45Ca deposition in aorta were measured as the index of calcification at the 8th and the 16th week.Results Compared with control group,the VDN group rats showed higher values of blood pressure from the 3rd week(all P<0.05).The ratio of heart to body weight increased 8.4%(P<0.01);plasma calcium content and plasma phosphate content increased 3.2%(P<0.01)and 12.0%(P<0.05),respectively;vascular calcium content increased 27.0%(P<0.01);vascular ALP activity and vascular calcium deposition increased 151.8% and 68.8%(P< 0.01),respectively.Compared with control group,at the 16th week,the ratio of heart to body weight increased 7.7%(P<0.01);the plasma calcium increased 3.2%(P<0.05),the plasma phosphate decreased 1.4%;vascular calcium content increased 51.0%;ALP activity and vascular calcium deposition increased 87.2%(P<0.01)and 31.6%(P<0.05),respectively.The result of Von Kossa staining shows many dark granules were found in the medial layer of aorta of VDN group rats at the 8th and the 16th week.Conclusion The sub-chronic 16 weeks rat model of vascular calcification induced by vitamin D3 and nicotine might be used for drug screenings and mechanism studies of diseases.