摘要
目的探讨柴胡软坚消瘿颗粒对肝郁脾虚型桥本甲状腺炎大鼠辅助性T细胞1(helper T cell 1,Th1)/辅助性T细胞2(helper T cell 2,Th2)、辅助性T细胞17(helper T cell 17,Th17)/调节性T细胞(regulatory T cell,Treg)平衡的影响。方法随机数字表法将48只大鼠分为正常组(12只)和造模组(36只),采用甲状腺球蛋白注射结合高碘饮水+慢性束缚+游泳过劳+饮食失节制备肝郁脾虚桥本甲状腺炎大鼠模型,共33只大鼠造模成功,将造模成功大鼠随机分为模型组、金水宝片[0.45 g/(kg·d)]和柴胡软坚消瘿颗粒组[11.43 g/(kg·d)],每组11只,连续给药8周。观察大鼠一般情况,实验前后记录大鼠体重、摄食量、粪便含水量,实验结束后旷场实验分析大鼠行为;末次给药后12小时,取血,流式细胞术检测Th1、Th2、Th17、Treg细胞百分比;酶联免疫吸附法检测大鼠血清甲状腺球蛋白抗体(thyroglobulin antibody,TGAb)、甲状腺过氧化酶抗体(thyroidperoxidase antibody,TPOAb)、游离三碘甲状腺原氨酸(free triiodothyronine,FT3)、游离甲状腺素(freethyroxine,FT4)、促甲状腺激素(thyroid stimulating hormone,TSH)、γ-干扰素(γ-interferon,IFN-γ)、白细胞介素10(interleukin 10,IL-10)、白细胞介素17(interleukin 17,IL-17)、白细胞介素35(interleukin35,IL-35)的水平;取血后分离大鼠甲状腺组织,苏木素—伊红染色法检测大鼠甲状腺组织病理形态;蛋白免疫印迹法检测大鼠甲状腺组织T盒子转录因子(T-box transcription factor,T-bet)、GATA结合蛋白3(GATA binding protein 3,GATA3)、叉头盒蛋白p3(forkhead box protein p3,Foxp3)、维A酸相关孤独受体γt(retinoid related orphan receptorγt,RORγt)的表达。结果与正常组相比,模型组大鼠甲状腺损伤严重,血清TPOAb、TGAb、FT3、FT4水平、Th1、Th17、Th1/Th2、Th17/Treg、IFN-γ、IL-17水平及蛋白T-bet、RORγt表达显著升高(P<0.01),TSH、Th2、Treg、IL-10、IL-35水平及蛋白GATA3、Foxp3表达显著降低(P<0.01);与模型组相比,金水宝片组、柴胡软坚消瘿颗粒组大鼠甲状腺损伤减轻,血清TPOAb、TGAb、FT3、FT4水平、Th1、Th17、Th1/Th2、Th17/Treg、IFN-γ、IL-17水平及蛋白T-bet、RORγt表达显著降低(P<0.01),TSH水平、Th2、Treg、IL-10、IL-35水平及蛋白GATA3、Foxp3表达显著升高(P<0.01)。结论柴胡软坚消瘿颗粒对肝郁脾虚桥本甲状腺炎大鼠治疗有一定的疗效,可能与调节Th1/Th2及Th17/Treg细胞平衡有关。
Objective To investigate the effect of Chaihu Ruanjian Xiaoying Granule on the balance of helper T cell 1(Th1)/helper T cell 2(Th2)and helper T cell 17(Th17)/regulatory T cell(Treg)in Hashimoto’s Thyroiditis(HT)rats with stagnation of liver-qi and deficiency of spleen type.Methods 48 rats were divided into normal group(12 rats)and model group(36 rats)by random number table.The rats in model group were injected with thyroglobulin and combined with high iodine drinking water,chronic restraint,swimming overwork,eating disorder.The HT rat model of stagnation of liver-qi and deficiency of spleen type were prepared.33 rats who were successfully modeled and then they were randomly divided into model group,Jinshuibao Tablet[0.45 g/(kg·d)]group and Chaihu Ruanjian Xiaoying Granule group[11.43 g/(kg·d)],11 rats per groupe.They were continuously administered for 8 weeks.The general condition of rats were observed,the weight,food intake,and fecal water content of were recorded during the experiment The behavior of rats was analyzed by the open field experiment;24 hours after the last administration,blood was taken,the percentages of Th1,Th2,Th17 and Treg cells were detected by the method of flow cytometry;the levels of thyroglobulin antibody(TGAb),thyroid peroxidase antibody(TPOAb),free triiodothyronine(FT3),free thyroxine(FT4),thyroid stimulating hormone(TSH),γ-interferon(IFN-γ),IL-10,IL-17,and IL-35 in serum were detected by enzyme-linked immunosorbent assay method;after blood collection,rat thyroid tissue was separated,and the pathological morphology of rat thyroid tissue was detected by hematoxylin-eosin staining method;the expression of T-box transcription factor(T-bet),GATA binding protein 3(GATA3),forkhead box protein p3(Foxp3)and retinoid related orphan receptorγt(RORγt)in rat thyroid tissue was detected by Western blotting.Results Compared with the normal group,the thyroid damaged rats in the model group was severe,the levels of TPOAb,TGAb,FT3,FT4,Th1,Th17,Th1/Th2,Th17/Treg,IFN-γ,IL-17 in serum and expressions of protein T-bet and RORγt were increased significantly,the levels of TSH,Th2,Treg,IL-10,IL-35 and the expressions of protein GATA3 and Foxp3 were reduced significantly(P<0.01);compared with the model group,rats in the Jinshuibao Tablet group and Chaihu Ruanjian Xiaoying Granule group had less thyroid injury,the levels of TPOAb,TGAb,FT3,FT4,Th1,Th17,Th1/Th2,Th17/Treg,IFN-γ,IL-17 in serum and expressions of protein T-bet and RORγt were reduced significantly,the levels of TSH,Th2,Treg,IL-10,IL-35 and the expressions of protein GATA3 and Foxp3 were increased significantly(P<0.01).Conclusion Chaihu Ruanjian Xiaoying Granule has effect on the treatment of HT rats of stagnation of liver-qi and deficiency of spleen type,which may be related to regulating the balance of Th1/Th2 and Th17/Treg cells.
作者
许可
张碧辰
商雪纯
张兰
XU Ke;ZHANG Bichen;SHANG Xuechun;ZHANG Lan(First School of Clinical Medicine,Liaoning University of Chinese Medicine,Shenyang 110032,China)
出处
《环球中医药》
CAS
2021年第10期1747-1753,共7页
Global Traditional Chinese Medicine
基金
辽宁省科学技术计划(2019-ZD-0967)。
关键词
柴胡软坚消瘿颗粒
肝郁脾虚型桥本甲状腺炎
辅助性T细胞1
辅助性T细胞2
辅助性T细胞17
调节性T细胞
Chaihu Ruanjian Xiaoying granule
Hashimoto’s thyroiditis of stagnation of liver-qi and deficiency of spleen type
Helper T cell 1
Helper T cell 2
Helper T cell 17
Regulatory T cell
