维生素D_3和华法令诱导的大鼠动脉钙化模型特点

Characteristics of rat models of arterial calcification induced by vitamin D_3 and warfarin

目的:探讨维生素D联合华法令对大鼠动脉钙化的作用,为心血管疾3病的康复预防措施介入提供基础研究数据。方法:实验选用12只4周龄雄性SD大鼠,随机分为钙化组和正常组,每组6只。钙化组给予皮下注射维生素D3×105U/(kg·d),持续3d,同3时华法令15mg/(100g·12h),持续4d,制备大鼠动脉钙化模型。取腹主动脉制成石蜡切片vonKossa染色观察动脉钙化情况,同时取胸主动脉按RT-PCR方法检测骨保护素的表达,另取右侧胫骨测量骨密度。结果:钙化组大鼠主动脉vonKossa染色见中膜有成片或局灶黑色深染区域,为主动脉的钙化部位。正常组主动脉壁骨保护素mRNA相对含量犤(45.6±0.6)%犦明显高于钙化组犤(24.6±0.4)%犦(P<0.05)。正常组大鼠胫骨骨密度犤(0.108±0.032)g/cm犦高于钙化组犤(0.0822±0.038)g/cm犦,但差异无显著性意义(P>0.05)。2结论:华法令和维生素D能诱导大鼠主动脉中膜钙化,钙化的主动脉3局部骨保护素的表达水平降低。

AIM:To study the calcification role of vitamin D3(VD3) combined with warfarin to the artery of rats,in order to provide basic studying data for the intervention of rehabilitative prevention measures to cardiovascular diseases. METHODS:Twelve 4 week old male SD rats were randomly divided into calcification group(n=6) and normal group(n=6).Rat models of arterial calcification in the calcification group were established by subcutaneous injection of VD3(3× 105 U/kg per day for 3 days) and warfarin (15 mg/100 g per 12 hours for 4 days).Abdominal aorta was collected for paraffin section, stained by von Kossa to observe the area of arterial calcification.The expressions of osteoprotegerin(OPG) in thoracic aorta were detected by reverse transcription polymerase chain reaction(RT PCR), and right tibias were taken to measure the bone mineral density(BMD). RESULTS:In the calcification group,large area or focal darkly stained regions, which were the aortic calcified sites, could be observed in tunica media of artery by von Kossa staining. The relative content of OPG mRNA in aortic wall was obviously higher in the normal group[(45.6± 0.6)% ] than in the calcification group[(24.6± 0.4)% ] (P< 0.05).The BMD of tibia in the normal group[(0.108± 0.032) g/cm2] was insignificantly higher than that in the calcification group[(0.082± 0.038) g/cm2] (P >0.05). CONCLUSION:Warfarin and VD3 can induce calcification of aortic tunica media in rat.The local OPG expression level in the calcified aorta is reduced.