Guo Ling, Qi Yong-fen
Department of Physiology, Peking University School of Basic Medical Sciences, Beijing 100083, China.
Beijing Da Xue Xue Bao Yi Xue Ban. 2005 Dec 18;37(6):656-8.
To investigate simple, economical, stable and efficient methods of in vivo and in vitro cardiovascular calcification models in rats.
Rats received Vitamin D(3) (300 000 U/kg, i.m.) and nicotine (25 mg/kg, 5 mL/kg in peanut oil, p.o.) at 8 a.m. on day 1. The nicotine administration was repeated at 6 p.m. Rats in control group received an injection of normal saline (i.m.) and two gavages of medium oil. All of the rats were then allowed to recover for 4 weeks and given standard rodent chow. After the measurement of cardiac function the rats were sacrificed and the calcium content in myocardium and aorta were measured. Von Kossa staining was used to detect the deposit of calcium in myocardium and aorta. Cultured smooth muscle cell(SMC) derived from rat thoracic aorta was treated with beta-glycerophosphate for 14 days, then the calcium content and deposit were measured.
Compared with control group, the rats with cardiovascular calcification showed a lower body weight, The ratio of left heart to body weight, myocardial and aortic calcium content were increased respectively. Alkaline phosphatases activity in calcified myocardium and aorta were increased respectively, compared with the control. The values of animal mean blood pressure (MBP), heart rate (HR) and left ventricle end-distolic pressure (LVEDP) showed no significant alteration (P>0.05) in vitamin D3 plus nicotine (VDN) group. The values of +LV dp/dt(max) and -LV dp/dtmax were significantly lower in VDN group (P<0.05 and P<0.01, respectively). In calcified vascular smooth muscle cells(VSMCs), von Kossa staining for calcification, showed positive staining as black/brown areas within the main, large, nodular structures as shown in extracellular matrix and cytoplasma. The content of calcium, (45)Ca(2+) uptake and alkaline phosphatase (ALP) activity in calcified VSMCs were increased (all P<0.01), respectively, compared with that of the control.
These methods can be used to produce calcification models in vivo and in vitro, which save money and time and are easy to manipulate.
研究简单、经济、稳定且高效的大鼠体内和体外心血管钙化模型构建方法。
第1天上午8点，大鼠接受维生素D3（300 000 U/kg，肌肉注射）和尼古丁（25 mg/kg，溶于5 mL/kg花生油中，口服）。下午6点重复给予尼古丁。对照组大鼠接受生理盐水肌肉注射和两次中链甘油三酯灌胃。然后让所有大鼠恢复4周并给予标准啮齿动物饲料。测量心脏功能后处死大鼠，测量心肌和主动脉中的钙含量。采用冯科萨染色法检测心肌和主动脉中的钙沉积。用β-甘油磷酸酯处理大鼠胸主动脉来源的培养平滑肌细胞（SMC）14天，然后测量钙含量和钙沉积。
与对照组相比，发生心血管钙化的大鼠体重较低，左心与体重之比、心肌和主动脉钙含量分别增加。钙化心肌和主动脉中的碱性磷酸酶活性分别高于对照组。维生素D3加尼古丁（VDN）组动物平均血压（MBP）、心率（HR）和左心室舒张末期压力（LVEDP）值无显著变化（P>0.05）。VDN组的+LV dp/dt（max）和-LV dp/dtmax值显著降低（分别为P<0.05和P<0.01）。在钙化的血管平滑肌细胞（VSMC）中，钙化的冯科萨染色显示在主要的大结节状结构内呈黑色/棕色区域的阳性染色，如细胞外基质和细胞质中所示。与对照组相比，钙化VSMC中的钙含量、（45）Ca（2+）摄取和碱性磷酸酶（ALP）活性均增加（均P<0.01）。
这些方法可用于构建体内和体外钙化模型，节省金钱和时间且易于操作。
