摘要
目的:通过观察中药复方软坚消瘿汤对实验性自身免疫性甲状腺炎(EAT)模型大鼠血清甲状腺球蛋白抗体(TgAb)、甲状腺过氧化酶抗体(TpoAb)、IL-1、6、TNF-α的影响及对甲状腺组织病理形态学、细胞凋亡相关蛋白Fas/FasL、Bcl-2/Bax的影响,从多靶点探讨软坚消瘿汤治疗慢性淋巴细胞性甲状腺炎的机理。
材料与方法:
以甲状腺球蛋白与弗氏完全佐剂连续两次皮下注射免疫实验大鼠,其间间隔两天,后三周以甲状腺球蛋白联合弗氏不完全佐剂对实验组大鼠再次进行皮下注射,以加强免疫,同时配合饮用高碘水(1L水中加入0.64 g碘化钠配制而成),获得实验性自身免疫性甲状腺炎(EAT)大鼠的动物模型。取实验组大鼠血清,检测Tg抗体的效价,随即将所有实验大鼠随机分为正常对照组、软坚消瘿汤治疗组、雷公藤多甙片对照组和模型组四组。软坚消瘿汤治疗6周后:(1)以酶联免疫法(ELISA法)检测各组大鼠血清TgAb、TpoAb、IL-1、6、TNF-α的变化;(2)以免疫组化法检测各组大鼠甲状腺组织中细胞凋亡相关蛋白Fas/FasL、Bcl-2/Bax的变化;(3)光镜下观察甲状腺组织病理形态学变化;(4)将各组结果进行统计学分析。
结果:
1.软坚消瘿汤治疗组与模型组比较:大鼠血清甲状腺自身抗体TgAb、TpoAb及IL-1、6、TNF-α水平明显降低,甲状腺组织Fas/FasL、Bax表达减少,甲状腺组织Bcl-2表达增加。光镜下观察软坚消瘿汤治疗组甲状腺组织结构损伤轻于模型组。
2.雷公藤多甙片治疗组与模型组比较:大鼠血清甲状腺自身抗体TgAb、TpoAb及IL-1、6、TNF-α水平明显降低,甲状腺组织Fas/FasL、Bax表达减少,甲状腺组织Bcl-2表达有所增加。光镜下观察雷公藤多甙片治疗组甲状腺组织结构损伤轻于模型组。
3.软坚消瘿汤治疗组与雷公藤多甙片对照组比较:软坚消瘿汤治疗组对各种指标的改善均优于雷公藤多甙片对照组。
结论:
1.软坚消瘿汤具有降低EAT大鼠血清甲状腺自身抗体及相关细胞因子水平的作用。
2.软坚消瘿汤具有减轻EAT大鼠甲状腺细胞凋亡的作用。
3.软坚消瘿汤具有减轻EAT大鼠甲状腺病理损伤的作用。
Purpose: To evaluate the effects of the traditional complex prescription Ruanjianxiaoying decoction in treatment of the EAT mice through the TGAb、TPOAb、IL-1、6、TNF-α、the pathomorphology and apoptosis-regulated proteins Fas/Fasl,Bcl-2/Bax change of the thyroid gland . Investigate the effect Mechanism from multiple target point.
Material and method:
An animal model of experimental autoimmune thyroiditis ( EAT) in rats was developed by injecting Tg and CFA two times,and the interval was two days. Then inject the mice with Tg and IFA for the next three weeks as strengthening the immune.Meanwhile,feed the EAT mice with high iodine water(1 liter water add to 0.64g NaI). So the model was completed . Take the experimental group serum to detect level of thyroglobulin antibody and then all the mice were divided into four groups randomly. They were normal group, Ruanjianxiaoying decoction treatment group , Glucosidorum Tripterygll Totorum treatment group and EAT group.Treat the mice for six weeks with Ruanjianxiaoying decoction:(1)Detect the levels of autoantibodies with Enzyme-linked immunosorbent assay.(2)Observe the level change of Fas/FasL,Bcl-2/Bax in the thyroid of rat with Immunohistochemical staining.(3)Survey the pathomorphology change of the thyroid gland under the light microscope(4)Compare the results between every groups.
Results:
1. Comparing the Ruanjianxiaoying decoction treatment group with the EAT group: the level of the TgAb、TpoAb、IL-1、6、TNF-α、Fas/Fasl and Bax were significantly lower.and the level of the Bcl-2 were higher. The pathomorphology change of the thyroid gland were better than that of the EAT group.
2. Comparing the Glucosidorum Tripterygll Totorum treatment group with the EAT group: the level of the TgAb、TpoAb、IL-1、6、TNF-α、Fas/Fasl and Bax were more lower.and the level of the Bcl-2 were higher. The pathomorphology change of the thyroid gland were better than that of the EAT group.
3. Comparing the Ruanjianxiaoying decoction treatment group with the Glucosidorum Tripterygll Totorum treatment group: the efficacy of the Ruanjianxiaoying decoction treatment group was better.
Conclusion:
1. Ruanjian Xiaoying Decoction has the function of lowering the expressions of antibodies and cytokines in serum levels of the EAT mice.
2. Ruanjianxiaoying Decoction has the function of reducing the apoptosis of the thyroid of the EAT mice.
3. Ruanjian Xiaoying Decoction has the function of alleviating the pathological injury of the thyroid of the EAT mice.
引文
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