阳和汤对自身免疫性甲状腺炎模型大鼠辅助性T细胞亚群的影响
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摘要
目的观察阳和汤对自身免疫性甲状腺炎大鼠辅助性T细胞(Th细胞)亚群平衡偏移的影响。方法 30只大鼠随机分为正常组、模型组和阳和汤组各10只,模型组和阳和汤组大鼠采用猪甲状腺球蛋白配合高碘饮水的方式建立自身免疫性甲状腺炎大鼠模型。造模后阳和汤组予阳和汤配方颗粒(药物浓度0.5 g/ml)1 ml/100 g灌胃,模型组及正常组予等量生理盐水灌胃,每日1次,各组连续灌胃10周。检测各组大鼠血清甲状腺过氧化物酶抗体(TPOAb)、甲状腺球蛋白抗体(Tg Ab)及甲状腺功能[包括促甲状腺激素(TSH)、血清游离三碘甲状腺原氨酸(FT3)、血清游离甲状腺素(FT4)]水平;脾脏提取T淋巴细胞,采用流式细胞检测技术测定Th1、Th2、Th17、调节性T细胞(Treg)表达情况,计算Th细胞亚群比例。结果与正常组比较,模型组和阳和汤组大鼠TSH明显降低,血清FT3、FT4、TPOAb、TgAb明显升高(P<0.01)。与模型组比较,阳和汤组大鼠FT4、TPOAb、TgAb明显降低(P<0.05或P<0.01)。与正常组比较,模型组Th1细胞占总淋巴细胞百分比、Th1/Th2、Th17占总淋巴细胞百分比、Th17/Treg升高,Th2细胞占总淋巴细胞百分比降低(P<0.05或P<0.01)。与模型组比较,阳和汤组Th1/Th2、Th17占总淋巴细胞百分比、Th17/Treg降低,但仍高于正常组(P<0.05或P<0.01)。各组大鼠Treg占总淋巴细胞百分比差异无统计学意义(P>0.05)。结论阳和汤可降低自身免疫性甲状腺炎模型大鼠甲状腺自身抗体滴度,降低大鼠脾脏Th17细胞百分比及Th1/Th2、Th17/Treg值,从而改善大鼠甲状腺自身免疫反应。
Objective To observe the effect of Yanghe Decoction( 阳和汤) on the balance shift of helper T cells( Th cell) subsets in rats with autoimmune thyroiditis. Methods Thirty rats were randomly divided into the normal group,the model group and Yanghe Decoction group,with 10 rats in each group. The model group and Yanghe Decoction group were treated with thyroglobulin plus drinking water to establish autoimmune thyroiditis rats model. After the modeling,the Yanghe Decoction group was given intragastric administration of Yanghe Decoction formula granules( concentration of drug 0. 5 g/ml) 1 ml/100 g. The model group and the normal group were intragastrically administrated with equal volume of saline,once a day,and each group was continuously administered intragastrically for 10 weeks. Serum thyroid peroxidase antibody( TPOAb),thyroglobulin antibody( Tg Ab) and thyroid function [including thyroid stimulating hormone( TSH),serum free triiodothyronine( FT3),serum free thyroid Levels of thymidine( FT4) ]were detected; T lymphocytes were extracted from the spleen and the expression of Th1,Th2,Th17 and regulatory T cells( Treg) was measured by flow cytometry,and the ratio of Th cell subsets was calculated. Results Compared with the normal group,the rats in the model group and Yanghe Decoction group had significantly lower TSH,and serum FT3,FT4,TPOAb,and Tg Ab were significantly higher( P < 0. 01). Compared with the model group,the FT4,TPOAb and Tg Ab in the Yanghe Decoction group were significantly lower( P < 0. 05 or P < 0. 01).Compared with the normal group,Th1 cells in the model group accounted for the percentage of total lymphocytes,Th1/Th2,Th17 as a percentage of total lymphocytes,Th17/Treg increased,and Th2 cells as a percentage of total lymphocytes decreased( P < 0. 05 or P < 0. 01). Compared with the model group,Th1/Th2,Th17 in the Yanghe Decoction group accounted for the percentage of total lymphocytes and Th17/Treg decreased,but still higher than the normal group( P < 0. 05 or P < 0. 01). There was no significant difference in total lymphocyte percentage between the Treg of each group( P > 0. 05). Conclusion Yanghe Decoction can reduce autoantibody titers of thyroid gland in rats with autoimmune thyroiditis,and reduce Th17 cell percentage and Th1/Th2,Th17/Treg in spleen of rats,and improve the thyroid autoimmune reaction in rats.
Objective To observe the effect of Yanghe Decoction( 阳和汤) on the balance shift of helper T cells( Th cell) subsets in rats with autoimmune thyroiditis. Methods Thirty rats were randomly divided into the normal group,the model group and Yanghe Decoction group,with 10 rats in each group. The model group and Yanghe Decoction group were treated with thyroglobulin plus drinking water to establish autoimmune thyroiditis rats model. After the modeling,the Yanghe Decoction group was given intragastric administration of Yanghe Decoction formula granules( concentration of drug 0. 5 g/ml) 1 ml/100 g. The model group and the normal group were intragastrically administrated with equal volume of saline,once a day,and each group was continuously administered intragastrically for 10 weeks. Serum thyroid peroxidase antibody( TPOAb),thyroglobulin antibody( Tg Ab) and thyroid function [including thyroid stimulating hormone( TSH),serum free triiodothyronine( FT3),serum free thyroid Levels of thymidine( FT4) ]were detected; T lymphocytes were extracted from the spleen and the expression of Th1,Th2,Th17 and regulatory T cells( Treg) was measured by flow cytometry,and the ratio of Th cell subsets was calculated. Results Compared with the normal group,the rats in the model group and Yanghe Decoction group had significantly lower TSH,and serum FT3,FT4,TPOAb,and Tg Ab were significantly higher( P < 0. 01). Compared with the model group,the FT4,TPOAb and Tg Ab in the Yanghe Decoction group were significantly lower( P < 0. 05 or P < 0. 01).Compared with the normal group,Th1 cells in the model group accounted for the percentage of total lymphocytes,Th1/Th2,Th17 as a percentage of total lymphocytes,Th17/Treg increased,and Th2 cells as a percentage of total lymphocytes decreased( P < 0. 05 or P < 0. 01). Compared with the model group,Th1/Th2,Th17 in the Yanghe Decoction group accounted for the percentage of total lymphocytes and Th17/Treg decreased,but still higher than the normal group( P < 0. 05 or P < 0. 01). There was no significant difference in total lymphocyte percentage between the Treg of each group( P > 0. 05). Conclusion Yanghe Decoction can reduce autoantibody titers of thyroid gland in rats with autoimmune thyroiditis,and reduce Th17 cell percentage and Th1/Th2,Th17/Treg in spleen of rats,and improve the thyroid autoimmune reaction in rats.
引文
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[5]邓莉,孙文,侯毅,等.自身免疫性甲状腺炎实验动物模型的研究进展[J].天津中医药,2016,33(3):189-192.
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[9]陶茂灿,贺倩倩,孙丹,等.阳和汤加减方治疗脾肾阳虚型系统性硬皮病临床疗效观察及机制探讨[J].浙江中医药大学学报,2016,40(6):445-450.
[10]WIERSINGA WM.Clinical relevance of environmental factors in the pathogenesis of autoimmune thyroid disease[J].Endocrinol Metab(Seoul),2016,31(2):213-222.
[11]KATJA Z,SIMONA G.Hashimoto's thyroiditis:from genes to the disease[J].Current Genomics,2011,12(8):576-588.
[12]RAMOS-LEV AM,MARAZUELA M.Pathogenesis of thyroid autoimmune disease:the role of cellular mechanisms[J].Endocrinol Nutr,2016,63(8):421-429.
[13]罗章伟.甲状腺自身抗体临床研究进展[J].右江医学杂志,2004,32(3):278-280.
[14]MCLACHLAN SM,RAPOPORT B.Why measure thyroglobulin autoantibodies rather than thyroid peroxidase autoantibodies?[J].Thyroid,2004,14(7):510-520.
[15]崔巍,施秉银.自身免疫性甲状腺病甲状腺内Th1/Th2平衡偏移研究[J].西安交通大学学报(医学版),2003,24(2):167.
[16]许峥嵘,李昭瑛.桥本甲状腺炎与Th1/Th2细胞因子[J].医学综述,2006,12(24):1492-1493.
[17]RAPOPORT B,MCLACHLAN SM.Graves'hyperthyroidism is antibody-mediated but is predominantly a Th1-type cytokine disease[J].J Clin Endocrinol Metab,2014,99(11):4060-4061.
[18]李志,杨婷婷,尹昂,等.桥本甲状腺炎患者外周血Th17与Treg细胞及其相关因子表达的研究[J].国际检验医学杂志,2013,34(8):945-946.
[2]TUNBRIDGE WM,VANDERPUMP MP.Population screening for autoimmune thyroid disease[J].Endocrinol Metab Clin North Am,2000,29(2):239-253.
[3]董笑克,邓莉,白颖,等.阳和汤对自身免疫性甲状腺炎大鼠甲状腺结构及甲状腺自身抗体的影响[J].环球中医药,2017,10(6):555-558.
[4]CUI SL,YU J,SHOUJUN L.Iodine intake increases IP-10 Expression in the serum and thyroids of rats with experimental autoimmune thyroiditis[J].Int J Endocrinol,2014,2014:581069.doi:10.1155/2014/581069.
[5]邓莉,孙文,侯毅,等.自身免疫性甲状腺炎实验动物模型的研究进展[J].天津中医药,2016,33(3):189-192.
[6]徐叔云,卞如濂,陈修.药理实验方法学[M].北京:人民卫生出版社,2003:1861
[7]王丹,邹国明,谌莉媚,等.喻文球临床新用阳和汤医案5则[J].江西中医药大学学报,2016,28(5):29-32.
[8]林丹丹,施红.施红教授应用阳和汤治疗系统性硬化病经验[J].福建中医药,2016,47(2):45-46.
[9]陶茂灿,贺倩倩,孙丹,等.阳和汤加减方治疗脾肾阳虚型系统性硬皮病临床疗效观察及机制探讨[J].浙江中医药大学学报,2016,40(6):445-450.
[10]WIERSINGA WM.Clinical relevance of environmental factors in the pathogenesis of autoimmune thyroid disease[J].Endocrinol Metab(Seoul),2016,31(2):213-222.
[11]KATJA Z,SIMONA G.Hashimoto's thyroiditis:from genes to the disease[J].Current Genomics,2011,12(8):576-588.
[12]RAMOS-LEV AM,MARAZUELA M.Pathogenesis of thyroid autoimmune disease:the role of cellular mechanisms[J].Endocrinol Nutr,2016,63(8):421-429.
[13]罗章伟.甲状腺自身抗体临床研究进展[J].右江医学杂志,2004,32(3):278-280.
[14]MCLACHLAN SM,RAPOPORT B.Why measure thyroglobulin autoantibodies rather than thyroid peroxidase autoantibodies?[J].Thyroid,2004,14(7):510-520.
[15]崔巍,施秉银.自身免疫性甲状腺病甲状腺内Th1/Th2平衡偏移研究[J].西安交通大学学报(医学版),2003,24(2):167.
[16]许峥嵘,李昭瑛.桥本甲状腺炎与Th1/Th2细胞因子[J].医学综述,2006,12(24):1492-1493.
[17]RAPOPORT B,MCLACHLAN SM.Graves'hyperthyroidism is antibody-mediated but is predominantly a Th1-type cytokine disease[J].J Clin Endocrinol Metab,2014,99(11):4060-4061.
[18]李志,杨婷婷,尹昂,等.桥本甲状腺炎患者外周血Th17与Treg细胞及其相关因子表达的研究[J].国际检验医学杂志,2013,34(8):945-946.