【作者】 龚惠琳；

【导师】 许芝银；

【作者基本信息】 南京中医药大学 ， 中医外科学（专业学位）， 2013， 博士

【摘要】 目的：观察雷公藤及其提取物对实验性自身免疫性甲状腺炎的作用效果及机理。方法：1.高碘水配制称取碘化钾0.64g溶于1L蒸馏水中,避光储存。2.分组造模及给药所有小鼠随机分为5组：即正常对照组(等量NS)、EAT模型组(等量NS)、雷公藤组(3.3g/kg)、雷公藤多甙片组(10mg/kg)、雷公藤甲素组(50μg/kg),每组10只。采用皮下多点注射TG抗原加高碘的方法制作EAT小鼠模型。除正常对照组外,各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。同时,各给药组按0.1ml/10g体重灌胃给药,正常对照组与模型对照组给予等体积生理盐水,末次给药后禁食24h。3.取材实验结束后小鼠眼眶取血,室温放置20min,2500r/min离心10min,取血清。采用放射免疫法测定T3、T4、FT3、FT4,TGAb、TMAb;甲状腺病理改变情况的影响；CD4+、 CD8+T淋巴细胞的比值的影响；甲状腺中Fas、Fas L改变情况的影响；甲状腺中caspase3改变情况的影响结果：1. EAT小鼠模型对照组血清T3、T4、FT3、FT4水平都高于正常组,差异有统计学意义(P<0.01)。与模型组比较,雷公藤、雷公藤多甙、雷公藤甲素三组的血清甲状腺激素都下降。雷公藤与雷公藤多甙两组的FT3,FT4,T3,T4下降存在显着性差异；雷公藤甲素组FT3,FT4,T4的下降存在显着性差异,T3差异性不显着。雷公藤、雷公藤多甙、雷公藤甲素三组的血清甲状腺激素都下降。雷公藤多甙组的TMAb、TGAb数值最低,下降最显着。2.模型组CD4+高于正常组、CD8+低于正常组,有显着性差异(p<0.01); CD4+/CD8+高于正常组,有显着性差异(p<0.01)。与模型组比较,雷公藤、雷公藤多甙、雷公藤甲素三组的CD4+比例下降、CD8+比例上升、CD4+/CD8+下降,且存在显着性差异。3.EAT小鼠模型组甲状腺中Fas、Fas-L表达水平都高于正常组,与正常组相比,差异有统计学意义(P<0.01)。与模型组比较,雷公藤、雷公藤多甙、雷公藤甲素三组的Fas、Fas-L都公藤多甙组的Fas、Fas-L数值最低,下降最显着。4.EAT小鼠模型组甲状腺中caspase3表达水平都高于正常组,与正常组相比,差异有统计学意义(P<0.01)。与模型组比较,雷公藤、雷公藤多甙、雷公藤甲素三组的caspase3都下降,存在显着性差异。雷公藤多甙组的caspase3数值最低,下降最显着。结论：本课题为雷公藤及其提取物临床应用于AT的治疗提供了实验依据,同时也为丰富治疗免疫性疾病中药的认识,扩展其应用提供了指导。随着人们对于AT的认识的不断深入以及中医药科学的不断发展,中药治疗AT将具有很大的发展空间和应用前景。更多还原

【Abstract】 PurposeInvestigate the effect and mechanism of Tripterygium and its extracts on experimental autoimmune thyroiditis.Method1. Preparation of Periodate water0.64g Potassium iodide dissolved in1L of distilled water, then storage it without sunlight reached.2. Group modeling and medicine administrationAll the small mice will separate into five groups randomly:Normal control group (striped NS), EAT model group (striped NS), Tripterygium group (3.3g/kg), Tripterygium Glycosides piece group (10mg/kg), Triptolide group (50μg/kg), n=10. Injected subcutaneously TG antigen plus periodate produced EAT mouse model. In addition to the normal control group, the mice in each group at the multiple sites subcutaneous injection of complete Freund’s adjuvant containing PTg (0.25mg/ml) antigen of0.2ml, two weeks in a row, once a week. The first three weeks of each mice were injected subcutaneously PTg (0.25mg/ml) incomplete Freund’s adjuvant antigen0.2ml, for three consecutive weeks, once a week. At the same time, each dose group0.1ml/10g weight is normal control group and model control group was given an equal volume of saline, fasted for24h after the last administration.3. CoverageAfter the end of the experiment, mice orbital blood will take out at room temperature for20min,2500r/min centrifugal10min, then take out the blood serum. Measured by radioimmunoassay T3, T4, FT3, FT4, TGAb, TMAb; thyroid pathological changes in the situation; CD4+, CD8+T lymphocyte ratio impact; thyroid in Fas, Fas L change the situation; thyroid caspase3change the impact of the situation.Result1. EAT mice model control group, blood serum T3, T4, FT3, FT4is higher than the normal group, the difference was statistically significant (P<0.01). Compared with model group, Tripterygium, Tripterygium glycosides, Tripterygium triptolide blood serum thyroid hormone are declined. Tripterygium and Tripterygium Glucosides two groups of FT3, FT4, T3, T4decline with a significant difference; Tripterygium Triptolide group FT3, FT4, the decline in the presence of T4significant difference, however the difference of T3was not significant. Wilfordii, Tripterygium glycosides, Tripterygium triptolide three groups of blood serum thyroid hormone declined. Tripterygium glycosides group TMAb, TGAb has the lowest value, which dropped the most significant.2. Model group, CD4+is higher than the normal group, CD8+is lower than the normal group, there is a significant difference (p<0.01); CD4+/CD8+is higher than the normal group, a significant difference (p<0.01). Group compared with the model group, Tripterygium, Tripterygium glycosides, Tripterygium Triptolide prime CD4+decline in the proportion, CD8+risen, CD4+/CD8+decreased, and there are significant differences.3. EAT mice model group thyroid Fas, Fas-L expression levels are higher than the normal group, compared with the normal group, the difference was statistically significant (P<0.01). Group compared with the model group, Tripterygium, Tripterygium glycosides, Tripterygium Triptolide prime Fas, Fas-L are public rattan polyglycoside group Fas, Fas-L lowest value, dropped the most significant.4. EAT mice model of thyroid caspase3expression levels are higher than the normal group, compared with the normal group, the difference was statistically significant (P<0.01). Group compared with the model group, Tripterygium, Tripterygium glycosides, Tripterygium Triptolide prime caspase3are down, there is a significant difference. Tripterygium glycosides the group caspase3numerical minimum, dropped the most significant.ConclusionThe subject is applied to the Tripterygium and its extract on clinical AT treatment to provide an experimental basis, as well as a rich understanding of Chinese medicine treatment of autoimmune diseases, extend its application to provide guidance. With the continuous development for people to further understand the AT as well as in pharmaceutical science, Chinese medicine treatment of AT will have more development spaces and application prospects.更多还原