维生素K对幼鼠骨量及骨强度影响的实验研究.doc

???? 维生素K对幼鼠骨量及骨强度影响的实验研究 ???? ???? Zhao Xinhua,Zhao Xihe,CuiWei,et alDepartment of biochemistry,Beijing Ji Shui Tan Hospital,Beijing 100035,China 　　Abstract Objective To explore the effect of various levels of vitamin K intake on bone development.Methods Forty weanling Wistar male rats were divided into four groups.In one group,1% sulfadiazine was added to regular diet(vitamin K 50 μg/kg) to induce vitamin K deficiency.In the other three groups,the vitamin K level in diets were 50,300 and 2550 μg/kg respectively.Twelve weeks later,the rats were killed and effects of the different level of vitamin K intake on bone development were evaluated by the parameters of bone metrology and bone biomechanics. Results Femur metrological parametres(bone weight,bone volume,bone mineral density and areas of trabecular bone etc.)improved with the increase of dietary vitamin K level,while the parametres of lumbar spine did not change when vitamin K level was over 300 μg/kg diet;vitamin K could improve the femoral structural biomechanical properties(strength and rigidity),but had no effect on the material biomechanical properties. Conclusion Vitamin K can enhance the development of bone.The rat vitamin K requirement may be higher than that currently recommended(50 μg/kg diet).　　Key words Vitamin K Bone metrology Bone biomechanics　　近年来，一些有关维生素K与骨代谢的研究提示维生素K能促进骨源性蛋白的羧化，但与骨发育的关系尚缺乏长期动物实验研究。骨计量学指标(骨重、骨密度、骨小梁面积等)可直观反映骨的再建情况及骨量多少，并且骨的力学性能是评价骨质量最有说服力的指标。本实验旨从骨计量学及骨生物力学等方面观察维生素K1对幼鼠骨发育的影响，以期从骨健康角度探讨维生素K的生理需要量。　　1　材料和方法1.1　动物来源及分组　　实验选用雄性Wistar断乳大鼠40只，体重(50±6)克，由中国医学科学院动物中心提供，按体重随机分为4组，分组情况如下：①Vk缺乏组(饲料中含1%磺胺嘧啶，阻断大鼠体内肠道菌群对Vk的合成)；②低剂量组(饲料含Vk 50 μg/kg)；③中剂量组(饲料含Vk300 μg/kg)；④高剂量组(饲料含Vk 2550 μg/kg)。1.2　动物喂养及饲料　　大鼠单笼饲养，自由进食，，饮去离子水，动物房温度保持在20～25℃，湿度50%左右，记录进食量，每周称体重一次，实验期3个月。饲料配方参照Joht T等的实验研究，经测定基础饲料Vk含量为50μg/kg，钙含量5.7 g/kg，缺乏组在基础饲料基础上添加1%磺胺嘧啶，低剂量组饲以基础饲料，中高剂量组分别在基础饲料中添加250μg和2500μg Vk，使分别达到每公斤饲料含Vk300和2550 mg(Vk为Sigma公司产品)。1.3　样品收集及处理　　实验结束前大鼠禁食一夜，次日晨在乙醚麻醉下由腹主动脉取血，摘取肝脏及双侧肾脏，用冷生理盐水冲洗，