半硫丸治疗实验性甲状腺功能减退症大鼠的作用及其机理研究
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摘要
目的 甲状腺功能减退症(甲减)是由于各种原因引起的甲状腺激素合成、分泌或生物效应不足所导致的体内代谢活动下降的一组临床综合征，是内分泌系统中较为常见的疑难病症。目前临床尚缺乏有效的根治方法。中医药治疗对本病有肯定疗效。为了积极寻求防治甲减的新的治疗方法，充分发挥中医药之整体调节、副作用小、疗效持久的优势，我们在半硫丸治疗甲减的临床基础上，以甲状腺功能减退症动物模型为实验对象，运用现代医学的理论和实验方法，对半硫丸治疗甲状腺功能减退症的作用及其机理进行了研究，从而为临床提供理论与实验依据。
方法 将受试大鼠随机平均分为6组：正常对照组、模型组、甲状腺片组、半硫丸小剂量组、半硫丸大剂量组、半硫丸加甲状腺片组。后五组予以造模，模型大鼠按1.35ml／200g体重／只，每日灌服0.04％他巴唑混悬液，共计造模45天。造模完成后开始灌服相应药物进行治疗实验，连续15天。然后各实验组分别进行如下指标检测：采用放射免疫法测定血清T_3、T_4、FT_3、FT_4、TSH及性激素(FSH、LH、E_2、P、T)水平；运用RT-PCR、Western blotting、Northern blotting，免疫组化等技术分别检测其海马T_3MRmRNA、Gs蛋白α亚基、Gi蛋白α亚基、Go蛋白α亚基的含量，GoαmRNA及SS的表达；光镜观察甲状腺和生殖器官的病理形态学变化，电镜观察甲状腺的超微结构。
结果 1．模型组在造模后出现甲减肾阳虚的行为学改变，反映甲减程度的临床指标T_3、T_4、FT_3、FT_4水平降低、TSH浓度升高，并且其甲状腺组织出现退行性的病理学改变。半硫丸尤其是半硫丸大剂量组不仅能改善实验大鼠的甲减症状，而且能明显升高T_3、T_4、FT_3、FT_4水平，降低TSH浓度，并可改善甲减大鼠甲状腺退行性的病理学改变；2．甲状腺功能减退症模型组大鼠血清性激素(FSH、LH、E_2、P、T)水平明显降低，大鼠生殖器官组织病理学均有明显退行性改变。半硫丸能显著提高实验性甲减大鼠血清性激素水平，改善其生殖器官组织病理形态学结构，且作用呈剂量依从性，大剂量组优于小剂量组；3．甲状腺功能减退症大鼠海马组织存在着T_3NRmRNA表达减弱，Gi蛋白α亚基、Go蛋白α亚基含量升高，GoαmRNA表达上调，SS表达增强等病理变化，不同剂量半硫丸对这些变化有不同程度的调整作用，其中半硫丸大剂量组效果最优。
结论 半硫丸能鼓舞阳气，促进机体的功能活动，对甲减大鼠具有良好的治疗作用，不仅能改善其甲减症状，升高血清甲状腺激素、性激素水平，减轻甲状腺、生殖系统等组织器官的退行性病理改变，而且能够调整甲减大鼠脑海马组织T_3NRmRNA，Gi蛋白α亚基、Go蛋白α亚基、GoαmRNA、SS的病理变化。半硫丸对甲减所引起的行为学的改变和生殖系统病变、甲减性脑损害多系统损伤的作用机理可能与增加内源性甲状腺激素分泌，减轻有害信号转导对机体的损害，调整机体神经内分泌功能紊乱等作用有关。
Objective Hypothyroidism is a common, difficult and complicated disease of endocrine system, which is a syndrome of hypometabolism caused by the deficiency of the synthesis, secretion and biological effect of thyroid hormone. Till now, there are no radical therapeutic methods for it in clinic. Chinese medicine may bring significant therapeutic effect on it. To take vigorous action on searching for a new therapeutic method for prevention and treatment of hypothyroidism by giving full play to the advantages of Traditional Chinese Medicine such as whole adjustment, less side effect, and long-term therapeutic effective, on the base of the clinical practice with Banliuwan, we study the function of Banliuwan on hypothyroidism and its mechanism under the basic theory and experimental methods of modern medicine with the hypothyroidism animal model in order to provide a theory and experimental base for the clinical practice.
Methods The rats were randomly divided into 6 groups, such as normal control group, model group, thyroxin group, small dosage group of Banliuwan, large dosage of Banliuwan, and the integrated group of thyroxin and Banliuwan. The rats of the latter 5 groups were made model by infusing 0.04% tapazole suspension 1.35ml/200g/day for 45 days. After that, the rats were treated with corresponding medicines for 15 days. Then the following detection were taken out: radioimmunoassay on serum FT3, FT4, T3, T4, TSH and sexual hormones (FSH, LH, E2, P, T); RT-PCR, Western blotting, Northern blotting and immunohistochemical method on the content or express of T3NRmRNA, Gsα, Gi α, Go α, Goα mRNA and SS; optical microscope and/or electron microscope observation on the pathology of thyroid and genitals.
Results (1) In the rats of model group, the behavior changes of kidney yang-insufficiency in hypothyroidism were appeared, and the indexes which may imply the severe degree of hypothyroidism presented the decreasing of FT3, FT4, T3 and T4, and the increasing of TSH, at the same time, the thyroid had retrograte changes in pathology. Banliuwan, especially with large dosage had not only improved the symptoms of hypothyroidism, but also increased the content of FT3, FT4, T3 and T4, and decreased the content of TSH, at the same time, it had improved the retrograte changes of the thyroid; (2) In the rats of model group, the level of serum sexual hormones (FSH, LH, E2, P, T) were obviously decreased and the genitals presented retrograte changes pathologically. Banliuwan significantly increased the content of sexual hormones and improved the structure of genitals. The effect depended on the dosage, that is, the effect of large dosage group was better than that of small dosage group. (3) In hippocampus of the hypothy
roidism rats, there were decreased expression of T3NRmRNA, increased expression or content of Gi α, Go α, Go α mRNA and SS. Different dosage of
Banliuwan brought different adjustment function on them, among which the best therapeutic effect was brought in large dosage group.
Conclusion Banliuwan, which may inspire yang-qi to promote the functional activity of the body, has good therapeutic effect on hypothyroidism rats with the improvement of symptoms, increasing of thyroid hormone and sexual hormone, improvement of the retrograte change of genitals and thyroid, as well as the adjustment of the pathological change of TaNRmRNA, Gi a , Go a, Go a mRNA and SS in hippocampus. The mechanism of Banliuwan on the symtom and the polysystem disorders of hypothyroidism, such as genital system and nervous system dysfunction, may be that Banliuwan may increase the endogenous thyroid hormone, reduce the injury by the damaged signal transduction, and adjust the neuroendocrine disorders.
方法 将受试大鼠随机平均分为6组：正常对照组、模型组、甲状腺片组、半硫丸小剂量组、半硫丸大剂量组、半硫丸加甲状腺片组。后五组予以造模，模型大鼠按1.35ml／200g体重／只，每日灌服0.04％他巴唑混悬液，共计造模45天。造模完成后开始灌服相应药物进行治疗实验，连续15天。然后各实验组分别进行如下指标检测：采用放射免疫法测定血清T_3、T_4、FT_3、FT_4、TSH及性激素(FSH、LH、E_2、P、T)水平；运用RT-PCR、Western blotting、Northern blotting，免疫组化等技术分别检测其海马T_3MRmRNA、Gs蛋白α亚基、Gi蛋白α亚基、Go蛋白α亚基的含量，GoαmRNA及SS的表达；光镜观察甲状腺和生殖器官的病理形态学变化，电镜观察甲状腺的超微结构。
结果 1．模型组在造模后出现甲减肾阳虚的行为学改变，反映甲减程度的临床指标T_3、T_4、FT_3、FT_4水平降低、TSH浓度升高，并且其甲状腺组织出现退行性的病理学改变。半硫丸尤其是半硫丸大剂量组不仅能改善实验大鼠的甲减症状，而且能明显升高T_3、T_4、FT_3、FT_4水平，降低TSH浓度，并可改善甲减大鼠甲状腺退行性的病理学改变；2．甲状腺功能减退症模型组大鼠血清性激素(FSH、LH、E_2、P、T)水平明显降低，大鼠生殖器官组织病理学均有明显退行性改变。半硫丸能显著提高实验性甲减大鼠血清性激素水平，改善其生殖器官组织病理形态学结构，且作用呈剂量依从性，大剂量组优于小剂量组；3．甲状腺功能减退症大鼠海马组织存在着T_3NRmRNA表达减弱，Gi蛋白α亚基、Go蛋白α亚基含量升高，GoαmRNA表达上调，SS表达增强等病理变化，不同剂量半硫丸对这些变化有不同程度的调整作用，其中半硫丸大剂量组效果最优。
结论 半硫丸能鼓舞阳气，促进机体的功能活动，对甲减大鼠具有良好的治疗作用，不仅能改善其甲减症状，升高血清甲状腺激素、性激素水平，减轻甲状腺、生殖系统等组织器官的退行性病理改变，而且能够调整甲减大鼠脑海马组织T_3NRmRNA，Gi蛋白α亚基、Go蛋白α亚基、GoαmRNA、SS的病理变化。半硫丸对甲减所引起的行为学的改变和生殖系统病变、甲减性脑损害多系统损伤的作用机理可能与增加内源性甲状腺激素分泌，减轻有害信号转导对机体的损害，调整机体神经内分泌功能紊乱等作用有关。
Objective Hypothyroidism is a common, difficult and complicated disease of endocrine system, which is a syndrome of hypometabolism caused by the deficiency of the synthesis, secretion and biological effect of thyroid hormone. Till now, there are no radical therapeutic methods for it in clinic. Chinese medicine may bring significant therapeutic effect on it. To take vigorous action on searching for a new therapeutic method for prevention and treatment of hypothyroidism by giving full play to the advantages of Traditional Chinese Medicine such as whole adjustment, less side effect, and long-term therapeutic effective, on the base of the clinical practice with Banliuwan, we study the function of Banliuwan on hypothyroidism and its mechanism under the basic theory and experimental methods of modern medicine with the hypothyroidism animal model in order to provide a theory and experimental base for the clinical practice.
Methods The rats were randomly divided into 6 groups, such as normal control group, model group, thyroxin group, small dosage group of Banliuwan, large dosage of Banliuwan, and the integrated group of thyroxin and Banliuwan. The rats of the latter 5 groups were made model by infusing 0.04% tapazole suspension 1.35ml/200g/day for 45 days. After that, the rats were treated with corresponding medicines for 15 days. Then the following detection were taken out: radioimmunoassay on serum FT3, FT4, T3, T4, TSH and sexual hormones (FSH, LH, E2, P, T); RT-PCR, Western blotting, Northern blotting and immunohistochemical method on the content or express of T3NRmRNA, Gsα, Gi α, Go α, Goα mRNA and SS; optical microscope and/or electron microscope observation on the pathology of thyroid and genitals.
Results (1) In the rats of model group, the behavior changes of kidney yang-insufficiency in hypothyroidism were appeared, and the indexes which may imply the severe degree of hypothyroidism presented the decreasing of FT3, FT4, T3 and T4, and the increasing of TSH, at the same time, the thyroid had retrograte changes in pathology. Banliuwan, especially with large dosage had not only improved the symptoms of hypothyroidism, but also increased the content of FT3, FT4, T3 and T4, and decreased the content of TSH, at the same time, it had improved the retrograte changes of the thyroid; (2) In the rats of model group, the level of serum sexual hormones (FSH, LH, E2, P, T) were obviously decreased and the genitals presented retrograte changes pathologically. Banliuwan significantly increased the content of sexual hormones and improved the structure of genitals. The effect depended on the dosage, that is, the effect of large dosage group was better than that of small dosage group. (3) In hippocampus of the hypothy
roidism rats, there were decreased expression of T3NRmRNA, increased expression or content of Gi α, Go α, Go α mRNA and SS. Different dosage of
Banliuwan brought different adjustment function on them, among which the best therapeutic effect was brought in large dosage group.
Conclusion Banliuwan, which may inspire yang-qi to promote the functional activity of the body, has good therapeutic effect on hypothyroidism rats with the improvement of symptoms, increasing of thyroid hormone and sexual hormone, improvement of the retrograte change of genitals and thyroid, as well as the adjustment of the pathological change of TaNRmRNA, Gi a , Go a, Go a mRNA and SS in hippocampus. The mechanism of Banliuwan on the symtom and the polysystem disorders of hypothyroidism, such as genital system and nervous system dysfunction, may be that Banliuwan may increase the endogenous thyroid hormone, reduce the injury by the damaged signal transduction, and adjust the neuroendocrine disorders.
引文
1．邱保国，王秀云，宁选，等．阳虚证型的转变与甲状腺关系的临床探讨．中西医结合杂志，1985；5(8)：479
2．张宏伟，陈如泉．半硫丸对家兔实验性甲减的治疗作用．中医药研究，1993；(3)：43
3．徐敏，邓响潮，张晓辉，等．巴戟滋补膏对甲状腺切除致阳虚兔血清甲状腺激素水平的影响．华西医大学报，1994；25(4)：431
4．秦路平，李博华，郑水庆，等．蛇床子素和淫羊藿甙的提取分离及补肾壮阳作用的研究．人民军医药学专刊，1997；13(4)：208
5．邝安堃，陈家伦，丁雯，等．益气助阳补肾中药对甲减模型动物组织形态学的影响．中国医药学报，1986；1(1)：22
6．邝安堃，丁雯，陈家伦，等．中医辨证论治对原发性甲状腺功能减退症的疗效及作用机理的初步探讨．中西医结合杂志，1983；3(6)：343
7．赵伟康，万叔援，蒋冰冰，等．温补肾阳药对甲状腺机能减退动物能量代谢的影响．中医杂志，1982；23(9)：58
8．任宏义，李浩澎，许凤云，等．肾阳虚证型病机学探讨—参附汤对甲状腺机能减退症大鼠的病理形态学研究．河南中医，1990；10(2)：37
9．徐敏，罗灼玲，张晓晖，等．巴戟滋补膏对甲低阳虚兔组织学变化的影响．新中医，1994；26(8)：63
10．罗灼玲，张立群，李文，等．温肾助阳药治疗甲状腺机能低下阳虚兔模型的组织学和组织化学观察．广州中医学院学报，1992；9(1)：45
11．陈汉平，胡国胜，何金森，等．对甲状腺机能减退阳虚证细胞免疫学的观察．上海中医药杂志，1989；(11)：36
12．庞洁，伍锐敏，曹淑兰．甲亢和甲减患者T淋巴细胞亚群的变化与中医的辩证关系．中日友好医院学报，1996；10(3)：261
13．刘桂梅，胡绍文，杨少敏，等，甲状腺功能减退症患者血液流变学异常的原因和意义．中华内科杂志，1988；27(10)：624
14．李瑞荃，王明芳，潘学会，等．“益视片”对实验性“肾阳虚”大鼠红细胞免疫功能的影响．成都中医药大学学报，1997；(1)：41
15．李文静，陈如泉．半硫丸对甲状腺机能减退肾阳虚大鼠血清SIL-2R水平影响的实验研究．辽宁中医学院学报，2002；4(1)：59
16．徐敏，张晓晖，欧明，等．温肾助阳药对甲低阳虚兔五种微量元素变化的影响．中国中医基础医学杂志，1995；1(3)：20
17．徐敏，张晓辉，欧敏，等．温肾补阳对甲低阳虚兔模型心、肾β受体的影响．福建中医药，1991；22(2)：30
18．赵胜利，刘红，刘洁，等．淫羊藿对甲状腺机能低下小鼠脑β受体和耗氧率的作用．河南医科大学学报，1996；(2)：45
19．徐敏，张晓辉，叶少梅，等．温肾助阳药对阳虚动物模型血液流变性的影响．中国医药学报，1995；10(2)：27
20．张家庆，丁光霞，刘福春，等．助阳药对阳虚动物脱氧核糖核酸合成率的作用．中医杂志，1982；23(9)：61
21．张家庆，刘福春，丁光霞，等．“阳虚”动物脱氧核糖核酸合成率和助阳药作用的研究．中医杂志，1982；23(3)：64
22．张善徵，徐敏，张晓晖．温补肾阳对甲状腺机能低下阳虚兔模型胰岛功能的影响．中医杂志，1988；(11)：63
23．徐敏，邓响潮，张晓辉，等．甲低兔血清皮质酮水平昼夜节律变化及助阳方药的影响．中药新药与临床药理，1995；6(4)：33
24．徐敏，邓响潮，张晓辉，等．温肾助阳方药对甲低阳虚模型体温和心率昼夜节律变化的影响．中国中药杂志，1996；21(4)：247
25．王琴琴，金静芬，蔡厚明，等．“甲减”患者肾阳虚与生长激素分泌变化的联系及温肾助阳的疗效．上海中医药杂志，1996；(6)：18
2．张宏伟，陈如泉．半硫丸对家兔实验性甲减的治疗作用．中医药研究，1993；(3)：43
3．徐敏，邓响潮，张晓辉，等．巴戟滋补膏对甲状腺切除致阳虚兔血清甲状腺激素水平的影响．华西医大学报，1994；25(4)：431
4．秦路平，李博华，郑水庆，等．蛇床子素和淫羊藿甙的提取分离及补肾壮阳作用的研究．人民军医药学专刊，1997；13(4)：208
5．邝安堃，陈家伦，丁雯，等．益气助阳补肾中药对甲减模型动物组织形态学的影响．中国医药学报，1986；1(1)：22
6．邝安堃，丁雯，陈家伦，等．中医辨证论治对原发性甲状腺功能减退症的疗效及作用机理的初步探讨．中西医结合杂志，1983；3(6)：343
7．赵伟康，万叔援，蒋冰冰，等．温补肾阳药对甲状腺机能减退动物能量代谢的影响．中医杂志，1982；23(9)：58
8．任宏义，李浩澎，许凤云，等．肾阳虚证型病机学探讨—参附汤对甲状腺机能减退症大鼠的病理形态学研究．河南中医，1990；10(2)：37
9．徐敏，罗灼玲，张晓晖，等．巴戟滋补膏对甲低阳虚兔组织学变化的影响．新中医，1994；26(8)：63
10．罗灼玲，张立群，李文，等．温肾助阳药治疗甲状腺机能低下阳虚兔模型的组织学和组织化学观察．广州中医学院学报，1992；9(1)：45
11．陈汉平，胡国胜，何金森，等．对甲状腺机能减退阳虚证细胞免疫学的观察．上海中医药杂志，1989；(11)：36
12．庞洁，伍锐敏，曹淑兰．甲亢和甲减患者T淋巴细胞亚群的变化与中医的辩证关系．中日友好医院学报，1996；10(3)：261
13．刘桂梅，胡绍文，杨少敏，等，甲状腺功能减退症患者血液流变学异常的原因和意义．中华内科杂志，1988；27(10)：624
14．李瑞荃，王明芳，潘学会，等．“益视片”对实验性“肾阳虚”大鼠红细胞免疫功能的影响．成都中医药大学学报，1997；(1)：41
15．李文静，陈如泉．半硫丸对甲状腺机能减退肾阳虚大鼠血清SIL-2R水平影响的实验研究．辽宁中医学院学报，2002；4(1)：59
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