【作者】 丁继存；

【导师】 许芝银；

【作者基本信息】 南京中医药大学 ， 中医外科学， 2010， 博士

【摘要】 目的麻雷汤是导师许芝银教授数十年治疗自身免疫性甲状腺炎临床有效方。本实验拟通过观察麻雷汤的急性毒性；观察麻雷汤对EAT小鼠脾淋巴细胞增殖的影响；观察麻雷汤对EAT小鼠外周血T淋巴细胞亚群的影响；观察麻雷汤对EAT小鼠甲状腺激素及自身抗体水平影响；观察麻雷汤对EAT小鼠甲状腺组织病理形态学的影响；观察麻雷汤对EAT小鼠甲状腺组织VEGF、Fas/Fas-L表达的影响。为临床应用该方提供充分的理论和准确的实验数据；探究麻雷汤治疗AT的可能作用机制；探讨AT的病因病机及治疗大法。方法麻雷汤的制备：麻黄300g,雷公藤300g,桃仁300g,肉桂300g。1200g生药煎成500ml水溶液,每ml含生药2.4g。取ICR小鼠50只,体重18.0-22.0g,随机分为5组,每组10只,雌雄各半,每组给药剂量分别为：26.88g/kg、33.6g/kg、42.Og/kg、53. Og/kg、67.Og/kg。实验前禁食(不禁水)12h,各组分别按上述剂量0.4ml／10g小鼠灌胃给药,1日1次,连续给药2周,观察记录实验小鼠活动、行为及死亡情况。取雌性ICR小鼠80只,体重18.0-22.0g,随机分为8组,每组10只。模型组小鼠于皮下多点注射含PTg(0.25mg／ml)完全弗氏佐剂抗原共0.2ml,连续2周,每周一次,第3周起每只小鼠皮下注射PTg(0.25mg/ml)不完全弗氏佐剂抗原0.2ml,连续3周,每周一次。除正常对照组外,其余各组小鼠每天给予饮用高碘水(1升水中加入0.64克碘化钾配制而成)。正常对照组和模型组均给予等容量生理盐水,甲状腺片组：10mg／kg,强的松组：10mg／kg,雷公藤组：25μg／kg,麻雷汤大剂量组：20g／kg；麻雷汤中剂量组：10g／kg；麻雷汤小剂量组：5g／kg。各组均灌胃给药,0.1ml／10g。造模2周后开始给药,连续4周,1日1次。每组取3只小鼠于末次给药后,摘眼球取血,处死,剪开腹膜取脾脏,观察EAT小鼠脾淋巴细胞增殖情况,流式细胞术检测外周血T淋巴细胞亚群。取雌性ICR小鼠160只,体重18.0-22.0g,随机分为8组,每组20只。处理如前。造模2周后开始给药,连续4周,1日1次。采用放射免疫法检测T3、T4、FT3、FT4、TSH、TGAb和TMAb;甲状腺组织作常规苏木素一伊红染色；观察小鼠甲状腺激素及自身抗体水平；免疫组化检测甲状腺组织VEGF、Fas/Fas-L蛋白的表达。结果麻雷汤经口给药,20只小鼠按96g／kg给药后均无异常情况发生,2周内亦无死亡,该剂量为成人日口服剂量的143倍(等效剂量的13倍)；模型组小鼠血清中高滴度甲状腺自身抗体TGAb.TMAb的检出(与正常组相比较P＜0.01),小鼠甲状腺组织的淋巴细胞浸润等表明成功复制出小鼠EAT模型；麻雷汤能抑制EAT小鼠脾淋巴细胞增殖(加或不加conA):调节其外周血T淋巴细胞亚群CD4+／CD8+；降低EAT小鼠甲状腺激素(T3、T4)及甲状腺自身抗体水平；减轻其甲状腺腺体扩张、破坏及淋巴细胞浸润；使EAT小鼠甲状腺组织VEGF及Fas／Fas-L低表达。结论麻雷汤是安全有效的治疗AT药物；麻雷汤抑制EAT小鼠脾淋巴细胞增殖；调节其外周血T淋巴细胞亚群CD4+／CD8+;降低其甲状腺激素(T3、T4)及甲状腺自身抗体水平；减轻其甲状腺腺体扩张、破坏及淋巴细胞浸润；降低其甲状腺组织VEGF及Fas／Fas-L表达。先天禀赋不足是本病的发病的基础和前提；脾肾阳虚是本病发生的主要病理阶段；气滞、痰凝、血瘀三者互结是本病发生的核心机制。脾肾阳虚、痰瘀互结是AT患者的主要病机；温阳破瘀化痰是本病的主要治疗大法。麻雷汤是脾肾阳虚、痰瘀互结型AT患者安全有效的经验方。更多还原

【Abstract】 Purpose:Ma Lei Tang is Professor Xu Zhiyin decades clinically effective treatment of autoimmune thyroiditis. In this study, by looking at Ma Lei Tang to be the acute toxicity; observation of Ma Lei Tang on EAT mice spleen lymphocyte proliferation; observation of Ma Lei Tang on EAT in mice peripheral blood T lymphocyte subsets; observation of Ma Lei Tang on EAT thyroid hormone and antibody levels in mice affected; observation of EAT in mice Ma Lei Tang on the impact of pathological thyroid tissue; observation of EAT in mice Ma Lei Tang on thyroid VEGF, Fas\Fas-L expression. For clinical application of the theory provided by full and accurate experimental data; explore the possible role of treatment Ma Lei Tang in AT mechanisms and Etiology, Pathogenesis and treatment of Dafa in AT.Methods:Preparation of Ma Lei Tang:Ephedra 300g, triptolide 300g, peach kernel 300g, cinnamon 300g.1200g crude drugs fry 500ml aqueous solution, each ml containing crude drugs 2.4g. ICR mice were obtained 50, weighing 18.0-22.Og, were randomly divided into five groups of 10 male and female in half, each dose were: 26.88g/kg,33.6 g/kg,42.0 g/kg,53.0 g/kg,67.0g/kg. Fasting before the experiment (can not help but water) 12h, respectively, in each group according to the above dose 0.4ml/10g mice gavage,1,1, continuous administration of 2 weeks, mice were observed and recorded activities, behavior and circumstances of death.80 female ICR mice were randomly divided into 8 groups of 10 mouse. Model group mice at the subcutaneous injection of multi-point PTg (0.25mg/ml) complete Freund’s adjuvant antigens were 0.2ml, for 2 weeks, once a week,3rd week of each mice were injected subcutaneously PTg (0.25mg/ml) incomplete Freund’s adjuvant antigen 0.2ml,3 consecutive weeks, once a week. In addition to the normal control group, the remaining mice in each group given a daily drink iodine water (1 liter in the preparation made by adding 0.64 grams potassium iodide). The normal control group and model group were given same volume of normal saline, thyroxine group: lOmg/kg, prednisone group:10mg/kg, tripterygium Group:25μg/kg, Ma Lei Tang high-dose group:20g/kg; Ma Lei Tang middle dose group:10g/kg; Ma Lei Tang low-dose group:5g/kg. In each group were gavage,0.1 ml/10g. Making model 2 weeks after administration for 4 weeks, qd. To take three mice in each group after the last administration, picking eye blood, death, cut to take the spleen peritoneum was observed EAT proliferation of mouse spleen lymphocytes by flow cytometry of peripheral blood T-lymphocyte subsets.160 female ICR mice were taken randomly divided into eight groups, each with 20. Treatment as before. Making model 2 weeks after administration for 4 weeks,1 day. Using radioimmunoassay T3, T4, FT3, FT4, TSH, TGAb and TMAb; thyroid tissue for routine hematoxylin-eosin stain; observation of thyroid hormone and antibody levels in mice; Immunohistochemical detection of thyroid tissue VEGF, Fas\ Fas-L protein expression.Results:Ma Lei Tang after oral administration,20 mice after administration by 96g/kg no exception happens, there is no death within 2 weeks, the dose for an adult day oral doses of 143 times (equivalent dose of 13 times); model group of high serum titer of thyroid autoantibodies TGAb, TMAb detection (compared with normal group P<0.01), infiltration of lymphocytes in thyroid tissue in mice, suggests that to replicate the success of mouse EAT model; Ma EAT Ray Tang can inhibit the proliferation of mouse spleen lymphocytes (with or without conA); adjust their peripheral blood T-lymphocyte subsets CD4+/CD8+; reduce the EAT in mice of thyroid hormone (T3, T4) and thyroid antibody levels; mitigate its thyroid gland expansion, destruction and lymphocyte infiltration; to EAT mice thyroid tissue VEGF and Fas, Fas-L with low expression.Conclusion:Ma Lei Tang is safe and effective treatment for AT drugs; Ma Lei Tang inhibited proliferation of mouse spleen lymphocytes EAT; adjust their peripheral blood T-lymphocyte subsets CD4+/CD8+; to reduce their thyroid hormone (T3, T4) and thyroid antibodies level; mitigate the thyroid gland expansion, destruction and lymphocyte infiltration; reduce their thyroid tissue VEGF and Fas, Fas-L expression. Congenital lack of disease incidence is the basis and prerequisite; spleen deficiency is Benbingfasheng The major pathological stage; Qi stagnation, Tanning, blood stasis three each node is the core mechanism Benbingfasheng. Spleen and kidney yang deficiency, phlegm and blood stasis is the main pathogenesis of patients with EAT; Onyang Po Yu phlegm is the main treatment of this disease Dafa. Ma Ray Tom is a spleen deficiency, phlegm and blood stasis type safe and effective in patients with AT experience side.更多还原