去卵巢骨質(zhì)疏松模型大鼠小腸鈣結(jié)合蛋白mRNA表達與菟絲子黃酮的干預(yù)

1、.去卵巢骨質(zhì)疏松模型大鼠小腸鈣結(jié)合蛋白mRNA表達與菟絲子黃酮的干預(yù)李小林1，武密山2，朱紫薇3，鄧勇存3，葉圓圓3，趙素芝4，任立中5，李  彬61河北醫(yī)科大學第二醫(yī)院，河北省石家莊市  050000；  2河北中醫(yī)學院基礎(chǔ)醫(yī)學院方劑學教研室，河北省石家莊市  050200；3河北中醫(yī)學院2011級中醫(yī)學專業(yè)，河北省石家莊市  050091； 4石家莊市橋東區(qū)勝北社區(qū)衛(wèi)生服務(wù)中心，河北省石家莊市  050041；5河北醫(yī)科大學第一醫(yī)院骨科，河北省石家莊市  050031；6河北醫(yī)科大學生物化學教研室，河北省石家莊

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：菟絲子是旋花科植物菟絲子Cuscutachinensis Lam.的成熟種子，為溫補腎陽的要藥，前期研究表明，由菟絲子組成的補腎復(fù)方在抑制骨量丟失，改善骨密度方面有明顯療效。目的：探討菟絲子黃酮對去卵巢骨質(zhì)疏松模型大鼠股骨骨密度、血清和腎臟1，25-二羥基維生素D3(1，25(OH)2 D3)含量、小腸鈣結(jié)合蛋白(CaBp-D9K)mRNA表達的影響。方法：72只SD雌性大鼠，隨機數(shù)字表法均分為6組(n=12

6、)：假手術(shù)組、模型組、維生素 D3組和菟絲子黃酮小、中、大劑量組。假手術(shù)組僅行假手術(shù)，其余5組分別行卵巢切除，1周后分別灌胃給予維生素D3 (2 mg/kg)以及小、中、大劑量菟絲子黃酮連續(xù)給藥3個月。腹主動脈取血，分離血清，取出腎臟，采用酶聯(lián)免疫吸附法檢測 1，25(OH)2 D3含量。之后處死動物，取出股骨，測定骨密度；取出第2腰椎，采用實時熒光反轉(zhuǎn)錄聚合酶鏈反應(yīng)(real-time RT-PCR)測定腰椎和腎臟組織維生素D受體mRNA表達。取出小腸，采用RT-PCR測定小腸CaBp-D9K mRNA表達。結(jié)果與結(jié)論：與假手術(shù)組相比，模型組股骨骨密度、血清和腎臟 1，25(OH)2 D3、

7、腰椎組織維生素D受體mRNA、小腸CaBp-D9K mRNA表達均下降。與模型組比較，菟絲子黃酮中、大劑量組和維生素D3組均可使股骨骨密度、血清和腎臟1，25(OH)2 D3、腰椎組織維生素D受體mRNA、小腸CaBp-D9K mRNA表達增加。菟絲子黃酮能夠顯著提高去卵巢大鼠股骨骨密度、血清和腎臟1，25(OH)2 D3、腰椎組織維生素D受體mRNA、小腸CaBp-D9K mRNA表達，促進腸鈣吸收與成骨細胞活性，增強骨質(zhì)量。中國組織工程研究雜志出版內(nèi)容重點：腎移植；肝移植；移植；心臟移植；組織移植；皮膚移植；皮瓣移植；血管移植；器官移植；組織工程全文鏈接：窗體頂端關(guān)鍵詞 ： 實驗動物； 組

8、織構(gòu)建； 骨質(zhì)疏松癥； 菟絲子黃酮； 骨密度；1； 25-二羥基維生素D3； 鈣結(jié)合蛋白； 大鼠； 國家自然科學基金；    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.中國組織工程研究雜志出版內(nèi)容重點：腎移植；肝移植；移植；心臟移植；組織移植；皮膚移植；皮瓣移植；血管移植；器官移植；組織工程全文鏈接：Key words： osteoporosis; bone density; flavones; calcium-binding proteins; osteoblasts; 窗體底端     中圖分類號: R318   基金資助: 國家自然科學基金資助項目(81073074，30472200)；河北省自然科學基金項目(H 2013206005)；河北省留學人員科技活動項目擇優(yōu)資助項目(200828)；河北省大學生創(chuàng)新創(chuàng)業(yè)訓練計劃項目(201310089062)通訊作者: 武密山，博士，教授，碩士生導師，河北中醫(yī)學院基礎(chǔ)醫(yī)學院方劑學教研室，河北省石家莊市 050200    作者簡介: 李小林，女，1957年生， 2003年河北醫(yī)科大學畢業(yè)，副主任醫(yī)師， 主要從事中藥治療風濕骨病作用機制研究。引用本文:   . 去卵巢骨質(zhì)疏松模型