肾阳虚骨质疏松大鼠动物模型的建立
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摘要
目的建立肾阳虚骨质疏松大鼠病证结合动物模型。方法大鼠按体质量随机分为3组,分别为实验组16只、去卵巢组16只及假手术组16只。实验组大鼠采用双侧去卵巢并im氢化可的松的方法进行造模。造模期间对大鼠一般情况进行观察及对生理状态量化检测。3个月后,采用ELISA法检测大鼠血清环磷酸腺苷(c AMP)、环磷酸鸟苷(c GMP)、雌二醇(E2)、睾酮(T)、促肾上腺皮质激素(ACTH)、三碘甲状腺原氨酸(T3)、四碘甲状腺原氨酸(T4)水平。用电子天平称肾上腺,甲状腺及胸腺湿质量,并取股骨进行骨密度和骨矿含量的检测。结果给药造模2周后,实验组大鼠出现毛发干枯、畏寒喜暖、卷缩弓背、小便清长等症状,体质量增长明显迟缓,血清c AMP水平较假手术组显著下降。3个月后,实验组大鼠自发活动次数明显减少,血清c AMP、E2、ACTH、T3、T4水平明显下降,骨密度、骨矿含量显著降低,肾上腺及甲状腺系数明显低于假手术组。结论去势并im氢化可的松建立肾阳虚骨质疏松大鼠动物模型是一种可行的方法。
Objective To establish an animal model of osteoporosis with kidney yang deficiency syndrome in rats. Methods Rats were randomly divided into three groups by body weight, They were experimental, ovariectomized, and Sham operation groups(16 in each group). In the experimental group, rats were subjected to bilateral ovarian resection followed by im injection of hydrocortisone. The general conditions and physiological conditions were observed and measured during the period of the experiment. After three months, the serum levels of cyclic adenosine monophosphate(c AMP), cyclic guanosine monophosphate(c GMP), estrogen(E2), testosterone(T), adrenocorticotropic hormone(ACTH), three typical thyroid original acid(T3), and four typical thyroid original acid(T4) were determined by ELISA method. The fresh weights of adrenal, thymus, and thyroid were collected by using electronic balance. In addition, the bone mineral density(BMD) and bone mineral content(BMC) were detected. Results Two weeks after hydrocortisone treatment, it was observed that the rats in the experimental group displayed kidney yang deficiency syndrome such as dry hairs, chills, back arched, and clear abundant urine. Correspondingly, the body weight increased slowly. The level of c AMP was significantly lower than that of rats in the Sham operation group. Three months later, spontaneous activities of rats in the experimental group markedly decreased, the content of c AMP, E2, ACTH, T3, and T4 significantly decreased when compared with the rats in the Sham operation group. The BMD and BMC decreased obviously, showing a notable statistic difference between the experimental group and Sham operation group. The coefficient of adrenal and thyroid was lower than that of rats in the Sham operation group. ConclusionBilateral ovarian resection followed by im injection of hydrocortisone may be a credible method for establishing the animal model of osteoporosis with kidney yang deficiency syndrome.
Objective To establish an animal model of osteoporosis with kidney yang deficiency syndrome in rats. Methods Rats were randomly divided into three groups by body weight, They were experimental, ovariectomized, and Sham operation groups(16 in each group). In the experimental group, rats were subjected to bilateral ovarian resection followed by im injection of hydrocortisone. The general conditions and physiological conditions were observed and measured during the period of the experiment. After three months, the serum levels of cyclic adenosine monophosphate(c AMP), cyclic guanosine monophosphate(c GMP), estrogen(E2), testosterone(T), adrenocorticotropic hormone(ACTH), three typical thyroid original acid(T3), and four typical thyroid original acid(T4) were determined by ELISA method. The fresh weights of adrenal, thymus, and thyroid were collected by using electronic balance. In addition, the bone mineral density(BMD) and bone mineral content(BMC) were detected. Results Two weeks after hydrocortisone treatment, it was observed that the rats in the experimental group displayed kidney yang deficiency syndrome such as dry hairs, chills, back arched, and clear abundant urine. Correspondingly, the body weight increased slowly. The level of c AMP was significantly lower than that of rats in the Sham operation group. Three months later, spontaneous activities of rats in the experimental group markedly decreased, the content of c AMP, E2, ACTH, T3, and T4 significantly decreased when compared with the rats in the Sham operation group. The BMD and BMC decreased obviously, showing a notable statistic difference between the experimental group and Sham operation group. The coefficient of adrenal and thyroid was lower than that of rats in the Sham operation group. ConclusionBilateral ovarian resection followed by im injection of hydrocortisone may be a credible method for establishing the animal model of osteoporosis with kidney yang deficiency syndrome.
引文
[1]周艳,扶晓明,贺栋梁,等.不同剂量葛根异黄酮联合Vit D对去势大鼠骨质疏松的防治[J].实用医学杂志,2009,25(23):3944-3946.
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[13]郑小伟,包素珍,刘明哲,等.金匮肾气丸对肾阳虚大鼠垂体ACTH基因表达的影响[J].中国中西医结合杂志,2004,24(3):238-240.
[14]沈自尹.有关证与神经内分泌免疫网络的研究[J].中医药学刊,2003,21(1):10-11.
[15]杨裕华,李震.肾阳虚动物模型及其诊断指标研究近况[J].辽宁中医药大学学报,2007,9(5):40-42.
[16]林燕萍,李咏高,王和鸣,等.健骨颗粒对骨质疏松模鼠垂体-甲状腺轴的影响[J].中国骨伤,2002,15(3):30-32.
[17]王建红,伍庆华,刘海云,等.肾阳虚大鼠垂体-甲状腺轴功能动态分析[J].辽宁中医杂志,2006,33(6):751-752.
[18]何成奇,谢薇,熊素芳,等.女性原发性骨质疏松肾虚三证与性激素变化关系的临床研究[J].华西医学,2003,18(1):14-15.
[19]苏成虎,杨新波,黄正明,等.水芹提取物对氢化可的松致肾阳虚小鼠的对抗作用[J].中国中医药信息杂志,2011,18(12):39-42.
[20]贾经汉,邱新建,陈志坚.骨质疏松动物模型的研究进展[J].中国康复医学杂志,2007,22(8):765-768.
[2]夏东胜,郭盛君,郭志强,等.绝经后妇女骨质疏松程度与肾虚证型的关系[J].新中医,2001,33(9):14-15.
[3]肖静,何立群,高建东,等.腺嘌呤与氢化可的松大鼠肾阳虚模型造模方法比较[J].中国比较医学杂志,2008,18(3):77-80.
[4]沈自尹,王文健.中医虚证辨证参考标准[J].临床荟萃,1987,(4):189.
[5]李跃华,董元龙.老年人骨质疏松患病率与肾虚证型关系的调查[J].中国中西医结合杂志,1995,15(6):366-367.
[6]胡永红,李丽春,梁燕,等.中医药治疗肾阳虚型骨质疏松症研究进展[J].河北中医药学报,2012,27(4):42-43.
[7]施明珠,李有贵,钟石,等.北冬虫夏草对氢化可的松诱导的老龄小鼠肾损伤的保护作用[J].现代药物与临床,2009,24(3):160-163.
[8]周文江,姚菊芳,彭秀华,等.肾阳虚证大鼠模型的建立[J].实验动物与比较医学,2007,27(4):242-243.
[9]富灵杰,汤亭亭,戴尅戎.骨质疏松动物模型的建立和应用[J].中华实验外科杂志,2008,25(2):270-272.
[10]钟历勇,沈自尹,蔡定芳,等.补肾健脾活血三类复方对下丘脑-垂体-肾上腺-胸腺轴及CRF基因表达的影响[J].中国中西医结合杂志,1997,17(1):39-41.
[11]李勇枝.中医肾虚证研究进展[J].中国中医基础医学杂志,1996,2(5):59-63.
[12]陈英华,欧阳轶强,孙琪,等.肾阳虚证动物模型规范化研究中诊断指标选择的初步探讨[J].中国中医基础医学杂志,2003,9(10):26-30.
[13]郑小伟,包素珍,刘明哲,等.金匮肾气丸对肾阳虚大鼠垂体ACTH基因表达的影响[J].中国中西医结合杂志,2004,24(3):238-240.
[14]沈自尹.有关证与神经内分泌免疫网络的研究[J].中医药学刊,2003,21(1):10-11.
[15]杨裕华,李震.肾阳虚动物模型及其诊断指标研究近况[J].辽宁中医药大学学报,2007,9(5):40-42.
[16]林燕萍,李咏高,王和鸣,等.健骨颗粒对骨质疏松模鼠垂体-甲状腺轴的影响[J].中国骨伤,2002,15(3):30-32.
[17]王建红,伍庆华,刘海云,等.肾阳虚大鼠垂体-甲状腺轴功能动态分析[J].辽宁中医杂志,2006,33(6):751-752.
[18]何成奇,谢薇,熊素芳,等.女性原发性骨质疏松肾虚三证与性激素变化关系的临床研究[J].华西医学,2003,18(1):14-15.
[19]苏成虎,杨新波,黄正明,等.水芹提取物对氢化可的松致肾阳虚小鼠的对抗作用[J].中国中医药信息杂志,2011,18(12):39-42.
[20]贾经汉,邱新建,陈志坚.骨质疏松动物模型的研究进展[J].中国康复医学杂志,2007,22(8):765-768.