去卵巢骨质疏松模型大鼠小肠钙结合蛋白mRNA表达与菟丝子黄酮的干预

1、.去卵巢骨质疏松模型大鼠小肠钙结合蛋白mRNA表达与菟丝子黄酮的干预李小林1，武密山2，朱紫薇3，邓勇存3，叶圆圆3，赵素芝4，任立中5，李  彬61河北医科大学第二医院，河北省石家庄市  050000；  2河北中医学院基础医学院方剂学教研室，河北省石家庄市  050200；3河北中医学院2011级中医学专业，河北省石家庄市  050091； 4石家庄市桥东区胜北社区卫生服务中心，河北省石家庄市  050041；5河北医科大学第一医院骨科，河北省石家庄市  050031；6河北医科大学生物化学教研室，河北省石家庄

2、市  050017Effects of flavonoids from Cuscuta chinensis on intestinal calcium-binding protein mRNA expression in ovariectomized osteoporosis model ratsLi Xiao-lin1, Wu Mi-shan2, Zhu Zi-wei3, Deng Yong-cun3, Ye Yuan-yuan3, Zhao Su-zhi4, Ren Li-zhong5, Li Bin61 Second Hospital of Hebei Medical Univ

3、ersity, Shijiazhuang 050000, Hebei Province, China; 2 Department of Prescriptions, Basic Medicine College, Hebei University of Traditional Chinese Medicine, Shijiazhuang 050200, Hebei Province, China; 3 Department of 2011 Grade of Traditional Chinese Medicine, Hebei University of Traditional Chinese

4、 Medicine, Shijiazhuang 050091, Hebei Province, China; 4 Qiaodong District Shengbei Community Health Center of Shijiazhuang, Shijiazhuang 050041, Hebei Province, China; 5 Department of Orthopedics, First Hospital, Hebei Medical University, Shijiazhuang 050031, Hebei Province, China; 6 Department of

5、Biochemistry, Hebei Medical University, Shijiazhuang 050017, Hebei Province, China摘要  背景：菟丝子是旋花科植物菟丝子Cuscutachinensis Lam.的成熟种子，为温补肾阳的要药，前期研究表明，由菟丝子组成的补肾复方在抑制骨量丢失，改善骨密度方面有明显疗效。目的：探讨菟丝子黄酮对去卵巢骨质疏松模型大鼠股骨骨密度、血清和肾脏1，25-二羟基维生素D3(1，25(OH)2 D3)含量、小肠钙结合蛋白(CaBp-D9K)mRNA表达的影响。方法：72只SD雌性大鼠，随机数字表法均分为6组(n=12

6、)：假手术组、模型组、维生素 D3组和菟丝子黄酮小、中、大剂量组。假手术组仅行假手术，其余5组分别行卵巢切除，1周后分别灌胃给予维生素D3 (2 mg/kg)以及小、中、大剂量菟丝子黄酮连续给药3个月。腹主动脉取血，分离血清，取出肾脏，采用酶联免疫吸附法检测 1，25(OH)2 D3含量。之后处死动物，取出股骨，测定骨密度；取出第2腰椎，采用实时荧光反转录聚合酶链反应(real-time RT-PCR)测定腰椎和肾脏组织维生素D受体mRNA表达。取出小肠，采用RT-PCR测定小肠CaBp-D9K mRNA表达。结果与结论：与假手术组相比，模型组股骨骨密度、血清和肾脏 1，25(OH)2 D3、

7、腰椎组织维生素D受体mRNA、小肠CaBp-D9K mRNA表达均下降。与模型组比较，菟丝子黄酮中、大剂量组和维生素D3组均可使股骨骨密度、血清和肾脏1，25(OH)2 D3、腰椎组织维生素D受体mRNA、小肠CaBp-D9K mRNA表达增加。菟丝子黄酮能够显著提高去卵巢大鼠股骨骨密度、血清和肾脏1，25(OH)2 D3、腰椎组织维生素D受体mRNA、小肠CaBp-D9K mRNA表达，促进肠钙吸收与成骨细胞活性，增强骨质量。中国组织工程研究杂志出版内容重点：肾移植；肝移植；移植；心脏移植；组织移植；皮肤移植；皮瓣移植；血管移植；器官移植；组织工程全文链接：窗体顶端关键词 ： 实验动物； 组

8、织构建； 骨质疏松症； 菟丝子黄酮； 骨密度；1； 25-二羟基维生素D3； 钙结合蛋白； 大鼠； 国家自然科学基金；    Abstract： BACKGROUND: Cuscuta chinensis is a mature seed of Cuscutachinensis Lam., can warm kidney. Previous studies demonstrated that kidney compound composed of Cuscuta chinensis could apparently inhibit bone loss and improve

9、 bone density.  OBJECTIVE: To explore the influence of flavonoids from Cuscuta chinensis on bone mineral density of femur, 1,25(OH)2D3 levels of serum and kidney, the expression of small intestine CaBp-D9K mRNA in model rats with ovariectomized osteoporosis.  METHODS: A total of 72 Sprague

10、-Dawley female rats were equally and randomly divided into six groups (n=12): sham surgery group, model group, vitamin D3 group and low-, moderate- and high-dose flavonoids groups. The sham surgery group only received sham operation and the other five groups were ovariectomized respectively. One wee

11、k after ovariectomy, the rats were given flavonoids from low-, moderate- and high-dose Cuscuta chinensis and vitamin D3 (2 mg/kg) by intragastric administration for 3 consecutive months. Blood was obtained from the abdominal aorta. Serum was isolated. The kidney was obtained. Enzyme linked immunosor

12、bent assay was used to determine 1,25(OH)2D3 contents of renal and serum. Rats were sacrificed at the end of experiment. The thighbone was taken out to determine bone mineral density. The second lumbar vertebra was taken out to measure the expression of lumbar vertebra and renal vitamin D receptor m

13、RNA using real-time fluorescent reverse transcription-polymerase chain reaction. The small intestine was taken out to measure the expression of small intestine CaBp-D9K mRNA using real-time fluorescent reverse transcription-polymerase chain reaction.RESULTS AND CONCLUSION: Compared with the sham ope

14、ration group, bone mineral density of femur, and 1,25(OH)2D3 levels of serum and kidney and the expression of lumbar vertebra vitamin D receptor mRNA significantly decreased in model group, and the expression of small intestine CaBp-D9K mRNA significantly decreased in model group. Compared with the

15、model group, bone mineral density of femur, and 1,25(OH)2D3 levels of serum and kidney, and the expression of the second lumbar vertebra vitamin D receptor mRNA, and the expression of small intestine CaBp-D9K mRNA were increased in moderate- and high-dose flavonoids groups and vitamin D3 group. Resu

16、lts indicated that flavonoids from Cuscuta chinensis could significantly raise bone mineral density of femur, and 1,25(OH)2D3 levels of serum and kidney and the expression of lumbar vertebra vitamin D receptor mRNA and the expression of small intestine CaBp-D9K mRNA, accelerate intestinal calcium ab

17、sorption and osteoblast activity, and reinforce quality of the bone.中国组织工程研究杂志出版内容重点：肾移植；肝移植；移植；心脏移植；组织移植；皮肤移植；皮瓣移植；血管移植；器官移植；组织工程全文链接：Key words： osteoporosis; bone density; flavones; calcium-binding proteins; osteoblasts; 窗体底端     中图分类号: R318   基金资助: 国家自然科学基金资助项目(81073074，30472200)；河北省自然科学基金项目(H 2013206005)；河北省留学人员科技活动项目择优资助项目(200828)；河北省大学生创新创业训练计划项目(201310089062)通讯作者: 武密山，博士，教授，硕士生导师，河北中医学院基础医学院方剂学教研室，河北省石家庄市 050200    作者简介: 李小林，女，1957年生， 2003年河北医科大学毕业，副主任医师， 主要从事中药治疗风湿骨病作用机制研究。引用本文:   . 去卵巢骨质疏松模型