碘诱发NOD和Balb/c小鼠甲状腺炎的实验研究

目的 观察碘过量对不同遗传背景小鼠甲状腺炎诱发作用的异同.方法 选用NOD和Balb/c小鼠各14只,每个品系小鼠均分为对照组、碘化钠(NaI)组,分别饮用纯净水和0.05%NaI水8周.放射免疫法(RIA)和ELISA法分别检测血清中总T4(TT4)、甲状腺球蛋白抗体(TgAb)、甲状腺过氧化物酶抗体(TPOAb)和促甲状腺激素(TSH)水平,光镜下观察甲状腺形态学改变和免疫组化染色的甲状腺细胞凋亡情况,四甲基偶氮唑兰(MTT)法检测颈部淋巴结和脾淋巴细胞对甲状腺球蛋白刺激的增殖反应.结果 饮碘水8周后,NOD和Balb/c小鼠甲状腺相对质量,NaI组[(104.83±14.52)、(155.79±20.77)mg/kg]较对照组[(71.80±20.42)、(105.15±21.98)mg/kg]明显增加(t值分别为-3.293、-4.429,P均<0.01),甲状腺组织表现为滤泡扩张、胶质潴留;NOD、Balb/c小鼠血清TT4水平,NaI组[(29.52±4.42)、(19.53±2.35)nmol/L]较对照组[(33.40±5.38)、(23.47±6.22)nmol/L]呈下降趋势(t值分别为1.374、1.567,P>0.05);血清TSH水平,与对照组[(3.55±1.41)、(5.55±0.56)μg/L]比较,Balb/c小鼠NaI组[(4.14±1.71)μg/L]呈升高趋势(t=-0.705,P>0.05),NOD小鼠NaI组[(6.98±0.66)μg/L]明显升高(t=-3.562,P<0.01);NOD、Balb/c小鼠NaI组TgAb(1281、1364 cpm)和TPOAb水平(2.50×103、0.14×103U/L)与对照组(1297、1220 cpm,3.17×103、0.03×103 U/L)比较没有明显变化(Z值分别为-0.081、-0.703,-0.244、-1.293,P均>0.05).NOD小鼠NaI组甲状腺细胞凋亡数量较BMb/c小鼠增加,并且甲状腺内可见大量淋巴细胞浸润,组织结构破坏和纤维化的灶性病变;NOD小鼠淋巴结和脾淋巴细胞数量,NaI组(1.100±0.014、1.076±0.033)较对照组(0.993±0.011、1.005±0.003)增加(t值分别为-11.672、-4.314,P均<0.01).结论 碘可致Balb/c鼠甲状腺肿大,功能下降;NOD鼠除七述病变外,甲状腺发生明显炎症反应,并且针对Tg产生了致敏的淋巴细胞.碘过量可诱发NOD鼠发生自身免疫性甲状腺炎.     

碘致NOD.H-2h4小鼠自身免疫甲状腺炎的实验研究

目的 探讨碘致自身免疫甲状腺炎发生过程中炎性细胞浸润和甲状腺自身抗体的动态变化.方法 NOD.H-2h4和昆明种小鼠各128只随机分入普通水组和0.05%碘化钠水组,分组后1、2、4、8、12、16、24周小鼠眶静脉取血,测定血清自身抗体和甲状腺激素,甲状腺组织制作石蜡切片,HE染色.结果 NOD.H-2h4小鼠饮用0.05%碘化钠水1周即有自身免疫性甲状腺炎的发生,补碘12周时患病率和炎性细胞浸润程度达最高峰.补碘8周,其血清TgAb开始升高,之后随着补碘时间的延长,其水平逐渐增高,而对照组血清TgAb始终保持低平曲线.结论 碘过量可以促进NOD.H-2h4小鼠自身免疫甲状腺炎(甲状腺淋巴细胞浸润)的发生和发展,血清TgAb是其炎症反应的一个标志.     

间日疟原虫重组蛋白Pvs25和Pvs28免疫小鼠应答机制观察

目的观察间日疟传播阻断疫苗候选抗原Pvs25和Pvs28的免疫应答机制。方法分别用与弗氏佐剂乳化后的Pvs25和Pvs28重组蛋白皮下免疫DBA/2小鼠;采用ELISA分别对各组小鼠脾细胞培养上清的IFN-γ、IL-4、IL-10动态检测;以ELISPOT方法检测小鼠脾脏IFN-γ+和IL-4+细胞数量。结果经重组蛋白免疫的两组小鼠脾细胞培养上清IFN-γ的含量于初次免疫后第14d升高,与对照组相比差异显著(P<0.01),但随后降至对照组水平;IL-4和IL-10均在初次免疫后第28d出现有意义的升高(P<0.01),后升高更为明显,分别直至免疫后期70d、56d出现下降,但直至112d仍明显高于免疫前的水平(P<0.01)。初次免疫后第70d?98d小鼠脾脏IFN-γ+和IL-4+细胞数量,与对照组相比IFN-γ+无明显变化,而IL-4+则明显增多(P<0.01)。结论Pvs25和Pvs28重组蛋白可诱导小鼠产生以Th2反应为主的免疫应答。     

高血压并冠状动脉粥样硬化与辅助性T淋巴细胞亚群

背景辅助性T细胞亚群中Th1细胞分泌干扰素(IFN-γ),Th2细胞分泌白介素4(IL-4),有报道认为动脉粥样硬化病人Th1/Th2平衡紊乱。目的探讨血管紧张素Ⅱ(AngⅡ),IFN-γ和IL-4与高血压患者冠状动脉粥样硬化的关系,研究他们的Th1/Th2功能平衡有无紊乱。方法临床及冠状动脉造影确诊的高血压合并冠状动脉粥样硬化患者30例为病例组,冠状动脉造影排除冠状动脉粥样硬化的高血压患者22例为对照组。采用放射免疫法检测血浆AngⅡ水平,ELISA法检测血浆中IFN-γ和IL-4水平。结果1)病例组AngⅡ[(64.7±30.1)vs对照组(36.1±12.0)ng/L,P<0.05]、IFN-γ[(10.3±5.4)vs(4.2±1.5)ng/L,P<0.01]水平增高;2)病例组IL-4水平无明显变化[(15.7±7.0)vs对照组(13.5±7.0)ng/L,P>0.05];3)病例组IFN-γ与IL-4比值较对照组增大(0.72±0.34vs0.35±0.17,P<0.01);4)AngⅡ水平与IFN-γ水平呈正相关(r=0.51,P<0.01),而与IL-4水平无相关性(r=0.11,P>0.05)。结论高血压患者的高AngⅡ水平使辅助性T淋巴细胞亚群Th1细胞活性升高,导致辅助性T淋巴细胞亚群Th1/Th2的功能失衡与冠状动脉粥样硬化发生、发展有关。     

碘过量和甲状腺球蛋白免疫诱发NOD小鼠甲状腺炎的病变特征

Objective To observe the pathological characteristics of thyroiditis induced by iodine excess and thyroglobulin (Tg) immunization and to explore the mechanism of thyroiditis induced by iodine excess. Methods NOD mice were used for intaking 0.05% Nal water and(or) Tg immunization. Morphologic change in thyroid and apoptosis were observed. The levels of serum TT4, TSH, thyroglobulin antibody (TgAb) and thyroid peroxidase antibody (TPOAb) were measured. Responding to Tg, lymphocytic proliferation of lymph node and spleen, interleukin-4(IL-4)and γ-interferon(IFN-γ) levels in culture medium of splenocytes were detected. Real-time PCR Was used to detect mRNA expressions of IL-4, IFN-γ, chemokine ligand 10 (CXCL10) and intercellular adhesion molecular-1(ICAM-1) in thyroid. Results Distended thyroid follicles,colloid accumulation, intense lymphocytic infiltration and disorganization were seen in thyroid of iodine excess group, along with increased apoptosis of thyroid cells(34.66～ 2.78 vs 5.11±0.62 ,P<0.01). The levels of TT4 were lowered while TSH raised ,but no production of thyroid-specific autoantibodies was revealed. Lymph node and spleen cells showed positive respornse under stimulation of Tg. The level of IFN-γ[(1. 272±0.049 vs 1. 139±0. 025)ng/L,P<0. 01] was raised in culture medium of splenocytes but not IL-4. The expression of IFN-γ, CXCLI0 and ICAM-1 mRNA were increased in thyroid. But in Tg group, some lymphocytes were scattered in thyroid, autoantibodies emerged ,and the level of IL-4 was increased in cuhure medium of splenocytes[(18. 508±0. 113 vs 13. 368±0. 016)ng/L, P<0. 01]. ledine excess combined with Tg enhanced these inflammatory reaction. Conclusion Iodine excess induced thyroiditis in NOD mice. The process seems to be Th1 response dominant organ-specific autoimmune diseases. Iodine excess and Tg immunizatiou play a synergistic role in inducing experimental autoimmune thyroiditis.     

碘过量和甲状腺球蛋白免疫诱发NOD小鼠甲状腺炎的病变特征

Objective To observe the pathological characteristics of thyroiditis induced by iodine excess and thyroglobulin (Tg) immunization and to explore the mechanism of thyroiditis induced by iodine excess. Methods NOD mice were used for intaking 0.05% Nal water and(or) Tg immunization. Morphologic change in thyroid and apoptosis were observed. The levels of serum TT4, TSH, thyroglobulin antibody (TgAb) and thyroid peroxidase antibody (TPOAb) were measured. Responding to Tg, lymphocytic proliferation of lymph node and spleen, interleukin-4(IL-4)and γ-interferon(IFN-γ) levels in culture medium of splenocytes were detected. Real-time PCR Was used to detect mRNA expressions of IL-4, IFN-γ, chemokine ligand 10 (CXCL10) and intercellular adhesion molecular-1(ICAM-1) in thyroid. Results Distended thyroid follicles,colloid accumulation, intense lymphocytic infiltration and disorganization were seen in thyroid of iodine excess group, along with increased apoptosis of thyroid cells(34.66～ 2.78 vs 5.11±0.62 ,P<0.01). The levels of TT4 were lowered while TSH raised ,but no production of thyroid-specific autoantibodies was revealed. Lymph node and spleen cells showed positive respornse under stimulation of Tg. The level of IFN-γ[(1. 272±0.049 vs 1. 139±0. 025)ng/L,P<0. 01] was raised in culture medium of splenocytes but not IL-4. The expression of IFN-γ, CXCLI0 and ICAM-1 mRNA were increased in thyroid. But in Tg group, some lymphocytes were scattered in thyroid, autoantibodies emerged ,and the level of IL-4 was increased in cuhure medium of splenocytes[(18. 508±0. 113 vs 13. 368±0. 016)ng/L, P<0. 01]. ledine excess combined with Tg enhanced these inflammatory reaction. Conclusion Iodine excess induced thyroiditis in NOD mice. The process seems to be Th1 response dominant organ-specific autoimmune diseases. Iodine excess and Tg immunizatiou play a synergistic role in inducing experimental autoimmune thyroiditis.     

碘过量和甲状腺球蛋白免疫诱发NOD小鼠甲状腺炎的病变特征

Objective To observe the pathological characteristics of thyroiditis induced by iodine excess and thyroglobulin (Tg) immunization and to explore the mechanism of thyroiditis induced by iodine excess. Methods NOD mice were used for intaking 0.05% Nal water and(or) Tg immunization. Morphologic change in thyroid and apoptosis were observed. The levels of serum TT4, TSH, thyroglobulin antibody (TgAb) and thyroid peroxidase antibody (TPOAb) were measured. Responding to Tg, lymphocytic proliferation of lymph node and spleen, interleukin-4(IL-4)and γ-interferon(IFN-γ) levels in culture medium of splenocytes were detected. Real-time PCR Was used to detect mRNA expressions of IL-4, IFN-γ, chemokine ligand 10 (CXCL10) and intercellular adhesion molecular-1(ICAM-1) in thyroid. Results Distended thyroid follicles,colloid accumulation, intense lymphocytic infiltration and disorganization were seen in thyroid of iodine excess group, along with increased apoptosis of thyroid cells(34.66～ 2.78 vs 5.11±0.62 ,P<0.01). The levels of TT4 were lowered while TSH raised ,but no production of thyroid-specific autoantibodies was revealed. Lymph node and spleen cells showed positive respornse under stimulation of Tg. The level of IFN-γ[(1. 272±0.049 vs 1. 139±0. 025)ng/L,P<0. 01] was raised in culture medium of splenocytes but not IL-4. The expression of IFN-γ, CXCLI0 and ICAM-1 mRNA were increased in thyroid. But in Tg group, some lymphocytes were scattered in thyroid, autoantibodies emerged ,and the level of IL-4 was increased in cuhure medium of splenocytes[(18. 508±0. 113 vs 13. 368±0. 016)ng/L, P<0. 01]. ledine excess combined with Tg enhanced these inflammatory reaction. Conclusion Iodine excess induced thyroiditis in NOD mice. The process seems to be Th1 response dominant organ-specific autoimmune diseases. Iodine excess and Tg immunizatiou play a synergistic role in inducing experimental autoimmune thyroiditis.     

碘过量和甲状腺球蛋白免疫诱发NOD小鼠甲状腺炎的病变特征

Objective To observe the pathological characteristics of thyroiditis induced by iodine excess and thyroglobulin (Tg) immunization and to explore the mechanism of thyroiditis induced by iodine excess. Methods NOD mice were used for intaking 0.05% Nal water and(or) Tg immunization. Morphologic change in thyroid and apoptosis were observed. The levels of serum TT4, TSH, thyroglobulin antibody (TgAb) and thyroid peroxidase antibody (TPOAb) were measured. Responding to Tg, lymphocytic proliferation of lymph node and spleen, interleukin-4(IL-4)and γ-interferon(IFN-γ) levels in culture medium of splenocytes were detected. Real-time PCR Was used to detect mRNA expressions of IL-4, IFN-γ, chemokine ligand 10 (CXCL10) and intercellular adhesion molecular-1(ICAM-1) in thyroid. Results Distended thyroid follicles,colloid accumulation, intense lymphocytic infiltration and disorganization were seen in thyroid of iodine excess group, along with increased apoptosis of thyroid cells(34.66～ 2.78 vs 5.11±0.62 ,P<0.01). The levels of TT4 were lowered while TSH raised ,but no production of thyroid-specific autoantibodies was revealed. Lymph node and spleen cells showed positive respornse under stimulation of Tg. The level of IFN-γ[(1. 272±0.049 vs 1. 139±0. 025)ng/L,P<0. 01] was raised in culture medium of splenocytes but not IL-4. The expression of IFN-γ, CXCLI0 and ICAM-1 mRNA were increased in thyroid. But in Tg group, some lymphocytes were scattered in thyroid, autoantibodies emerged ,and the level of IL-4 was increased in cuhure medium of splenocytes[(18. 508±0. 113 vs 13. 368±0. 016)ng/L, P<0. 01]. ledine excess combined with Tg enhanced these inflammatory reaction. Conclusion Iodine excess induced thyroiditis in NOD mice. The process seems to be Th1 response dominant organ-specific autoimmune diseases. Iodine excess and Tg immunizatiou play a synergistic role in inducing experimental autoimmune thyroiditis.     

碘过量和甲状腺球蛋白免疫诱发NOD小鼠甲状腺炎的病变特征

Objective To observe the pathological characteristics of thyroiditis induced by iodine excess and thyroglobulin (Tg) immunization and to explore the mechanism of thyroiditis induced by iodine excess. Methods NOD mice were used for intaking 0.05% Nal water and(or) Tg immunization. Morphologic change in thyroid and apoptosis were observed. The levels of serum TT4, TSH, thyroglobulin antibody (TgAb) and thyroid peroxidase antibody (TPOAb) were measured. Responding to Tg, lymphocytic proliferation of lymph node and spleen, interleukin-4(IL-4)and γ-interferon(IFN-γ) levels in culture medium of splenocytes were detected. Real-time PCR Was used to detect mRNA expressions of IL-4, IFN-γ, chemokine ligand 10 (CXCL10) and intercellular adhesion molecular-1(ICAM-1) in thyroid. Results Distended thyroid follicles,colloid accumulation, intense lymphocytic infiltration and disorganization were seen in thyroid of iodine excess group, along with increased apoptosis of thyroid cells(34.66～ 2.78 vs 5.11±0.62 ,P<0.01). The levels of TT4 were lowered while TSH raised ,but no production of thyroid-specific autoantibodies was revealed. Lymph node and spleen cells showed positive respornse under stimulation of Tg. The level of IFN-γ[(1. 272±0.049 vs 1. 139±0. 025)ng/L,P<0. 01] was raised in culture medium of splenocytes but not IL-4. The expression of IFN-γ, CXCLI0 and ICAM-1 mRNA were increased in thyroid. But in Tg group, some lymphocytes were scattered in thyroid, autoantibodies emerged ,and the level of IL-4 was increased in cuhure medium of splenocytes[(18. 508±0. 113 vs 13. 368±0. 016)ng/L, P<0. 01]. ledine excess combined with Tg enhanced these inflammatory reaction. Conclusion Iodine excess induced thyroiditis in NOD mice. The process seems to be Th1 response dominant organ-specific autoimmune diseases. Iodine excess and Tg immunizatiou play a synergistic role in inducing experimental autoimmune thyroiditis.     

碘过量和甲状腺球蛋白免疫诱发NOD小鼠甲状腺炎的病变特征

Objective To observe the pathological characteristics of thyroiditis induced by iodine excess and thyroglobulin (Tg) immunization and to explore the mechanism of thyroiditis induced by iodine excess. Methods NOD mice were used for intaking 0.05% Nal water and(or) Tg immunization. Morphologic change in thyroid and apoptosis were observed. The levels of serum TT4, TSH, thyroglobulin antibody (TgAb) and thyroid peroxidase antibody (TPOAb) were measured. Responding to Tg, lymphocytic proliferation of lymph node and spleen, interleukin-4(IL-4)and γ-interferon(IFN-γ) levels in culture medium of splenocytes were detected. Real-time PCR Was used to detect mRNA expressions of IL-4, IFN-γ, chemokine ligand 10 (CXCL10) and intercellular adhesion molecular-1(ICAM-1) in thyroid. Results Distended thyroid follicles,colloid accumulation, intense lymphocytic infiltration and disorganization were seen in thyroid of iodine excess group, along with increased apoptosis of thyroid cells(34.66～ 2.78 vs 5.11±0.62 ,P<0.01). The levels of TT4 were lowered while TSH raised ,but no production of thyroid-specific autoantibodies was revealed. Lymph node and spleen cells showed positive respornse under stimulation of Tg. The level of IFN-γ[(1. 272±0.049 vs 1. 139±0. 025)ng/L,P<0. 01] was raised in culture medium of splenocytes but not IL-4. The expression of IFN-γ, CXCLI0 and ICAM-1 mRNA were increased in thyroid. But in Tg group, some lymphocytes were scattered in thyroid, autoantibodies emerged ,and the level of IL-4 was increased in cuhure medium of splenocytes[(18. 508±0. 113 vs 13. 368±0. 016)ng/L, P<0. 01]. ledine excess combined with Tg enhanced these inflammatory reaction. Conclusion Iodine excess induced thyroiditis in NOD mice. The process seems to be Th1 response dominant organ-specific autoimmune diseases. Iodine excess and Tg immunizatiou play a synergistic role in inducing experimental autoimmune thyroiditis.     

碘致甲状腺滤泡上皮细胞损伤在甲状腺炎发生机制中的探讨

目的 探讨过量碘对甲状腺滤泡上皮超微结构的影响,从而探讨甲状腺损伤与自身免疫甲状腺炎之间的关系.方法 NOD.H-2h4和昆明小鼠随机分人饮用正常水组和5倍、10倍、100倍高碘水组.各组动物分别于实验开始后4、8及24周处死,锇酸固定,醋酸双氧铀和柠檬酸铅双重染色,透射电镜成像.结果碘过量造成NOD.H-2h4小鼠甲状腺上皮细胞过氧体和次级溶酶体数量增多、上皮细胞凋亡、坏死、滤泡结构破坏和炎细胞的浸润,而且这种损伤与碘剂量呈正相关.结论 碘过量对甲状腺上皮细胞的氧化损伤作用可能是自身免疫甲状腺炎产生的必要条件.     

碘过量对NOD和Balb/c鼠甲状腺TRAIL和TRAIL-sR1表达的影响

目的 观察碘过量对自身免疫性疾病敏感品系NOD鼠和非敏感品系Balb/c鼠甲状腺细胞凋亡相关基因肿瘤坏死因子的相关凋亡诱导配体(tumor necrosis factor-related apoptosis-inducing ligand, TRAIL)和其受体TRAIL刺激性受体1(TRAIL-sR1)表达的影响,了解碘过量诱导实验性自身免疫性甲状腺炎(EAT)发病过程中TRAIL和TRAIL-sR1的作用机制.方法 6～7周龄雌性NOD鼠和Balb/c鼠各16只,按体质量各随机分为对照组和碘过量(HI)组,每组8只.对照组饮高压灭菌水,HI组饮0.05%NaI添加水,饲养8周后处死.测量甲状腺相对质量,进行甲状腺组织形态学观察,测量血清甲状腺素(TT4)和甲状腺刺激激素(TSH),检测甲状腺滤泡上皮细胞凋亡情况,应用实时定量PcR(real time PCR)方法检测TRAIL和TRAIL-sR1 mRNA表达情况.结果 NOD鼠HI组甲状腺相对质量[(104.8±14.5)mg/kg]高于对照组[(71.8±20.4)mg/kg],血清TT4水平[(30.77±3.59)mmol/L]低于对照组[(36.43±2.66)mmol/L],TSH水平[(6.98±0.66)μg/L]高于对照组[(5.55±0.56)μg/L],组间比较差异均有统计学意义(t值分别为7.773、-9.526、-4.458,P均<0.05).HI组甲状腺出现滤泡扩张、胶质潴留及明显的淋巴细胞浸润伴灶性纤维化.Balb/c鼠川组甲状腺相对质量[(155.8±20.8)mg/kg]高于对照组[(105.1±22.0)mg/kg],血清TT4水平[(19.75±3.32)mmol/L]低于对照组[(23.46±6.21)mmol/L],TSH水平[(4.14±1.71)μg/L]高于对照组[(3.55±1.41)μg/L],组间比较差异均有统计学意义(t值分别为7.554、-7.239、3.140,P均<0.05);HI组甲状腺仅有甲状腺滤泡扩张,胶质潴留,而未见明显的淋巴细胞浸润.HI组NOD鼠和Balb/c鼠甲状腺滤泡上皮细胞的凋亡指数(3.97±0.91、1.05±0.45)分别高于对照组(0.21±0.15、0.10±0.03),组间比较差异均有统计学意义(t值分别为-7.167、-17.772,P均<0.05),Balb/c鼠HI组的甲状腺滤泡上皮细胞的凋亡指数低于NOD鼠HI组,组间比较差异有统计学意义(t=-7.625,P<0.05).NOD鼠HI组TRAIL mRNA表达水平(0.018 88±0.005 77)高于对照组(0.00961±0.005 91),组间比较差异有统计学意义(t=-2.710,P<0.05);Balb/c鼠HI组TRAIL表达水平(0.001 24±0.000 46)与对照组(0.000 59±0.000 39)比较,差异无统计学意义(t=-1.940,P>0.05);NOD鼠和Balb/c鼠HI组的TRAIL-sR1 mRNA表达水平(0.000 53±0.000 15、0.000 42±0.000 09)均高于对照组(0.000 28±0.000 05、0.000 17±0.000 06),组间比较差异有统计学意义(t值分别为3.050,3.990,P均<0.05),Balb/c鼠HI组的TRAIL和TRAIL-sR1的mRNA表达水平均低于NOD鼠HI组,组间比较差异均有统计学意义(t值分别为-3.370、-4.760,P均<0.05).结论 碘过量可使NOD和Balb/c鼠发生胶质潴留性甲状腺肿,并可造成NOD鼠甲状腺发生明显的炎症反应.TRAIL和TRAIL-sR1表达水平升高是碘过量导致甲状腺滤泡上皮凋亡和炎症发生的分子基础之一.遗传在甲状腺炎的发病过程中起到至关重要的作用.     

Induction of autoimmune thyroiditis by depletion of CD4+CD25+ regulatory T cells in thyroiditis-resistant IL-17, but not interferon-gamma receptor, knockout nonobese diabetic-H2h4 mice

Iodine-induced experimental autoimmune thyroiditis in the nonobese diabetic (NOD)-H2h4 mouse is a prototype of animal models of Hashimoto's thyroiditis in humans. Recent studies have shown the resistance to thyroiditis of NOD-H2h4 mice genetically deficient for either IL-17 or interferon (IFN)-γ, implicating both of T helper type 1 (Th1) and Th17 immune responses in disease pathogenesis. However, we hypothesized that robust induction of a single arm of effector T cells (either Th1 or Th17) might be sufficient for inducing thyroiditis in NOD-H2h4 mice. To address this hypothesis, enhanced immune responses consisting of either Th1 or Th17 were induced by anti-CD25 antibody-mediated depletion of regulatory T cells (Treg) in thyroiditis-resistant IL-17 knockout (KO) or IFN-γ receptor (IFN-γR) KO, respectively, NOD-H2h4 mice. Depletion of Treg in IL-17 KO mice (i.e. Th1 enhancement) elicited antithyroglobulin autoantibodies and thyroiditis. Immunohistochemical analysis of the thyroid glands revealed the similar intrathyroidal lymphocyte infiltration patterns, with CD4+ T and CD19+ B cells being dominant between the wild-type and Treg-depleted IL-17 KO mice. In contrast, Treg-depleted IFN-γR KO mice remained thyroiditis resistant. Intracellular cytokine staining assays showed differentiation of Th1 cells in IL-17 KO mice but not of Th17 cells in IFN-γR KO mice. Our findings demonstrate that a robust Th1 immune response can by itself induce thyroiditis in otherwise thyroiditis-resistant IL-17 KO mice. Thus, unlike Th17 cells in IFN-γR KO mice, Th1 cells enhanced by Treg depletion can be sustained and induce thyroiditis.     

碘摄入量对血清甲状腺球蛋白的影响——5年流行病学随访研究

目的 观察碘摄入量对血清甲状腺球蛋白(Tg)水平的影响.方法 对1999年盘山(轻度碘缺乏地区)、彰武(碘超足量地区)和黄骅(碘过量地区)血清Tg水平正常的3 099人进行随访,将2004年随访到的2 448人作为研究人群.测定Tg、甲状腺球蛋白抗体(TgAb)、甲状腺过氧化物酶抗体(TPOAb)、促甲状腺素(TSH)、甲状腺容积,并询问甲状腺疾病的家族史和个人史.分析Tg的影响因素.结果 1999年初访时,不同碘摄入地区Tg水平差异显著[盘山7.5(4.4～13.1)μg/L、黄骅6.8(3.6～11.2)μg/L彰武5.9(3.2～10.7)μg/L,P<0.01];年龄、性别、TgAb阳性、甲状腺肿大、TSH异< 凹鬃聪偌膊「鋈耸范訲g水平的影响亦显著.为排除混杂因素的影响,对TgAb、甲状腺容积、TSH、甲状腺疾病个人史均无异常的1 856人的随访数据进行分析,其初访及随访的Tg水平均有显著地区差异,轻度碘缺乏以及碘过量地区都显著高于碘超足量地区.三地区随访时Tg水平均显著升高,且增加值(△Tg)存在显著地区差异[盘山3.1(-0.2～8.0)μg/L、黄骅3.5(0.5～9.0)μg/L/35彰武2.5(0.3～6.1)μg/L,P<0.01];碘摄入量、初访时的Tg水平、TSH水平及年龄为ATg的独立影响因素.结论 碘摄入量显著影响血清Tg水平,当用血清Tg水平来估计地区碘摄入水平及其变化时,应该同时考虑年龄及TSH等因素.     

碘对小鼠产后甲状腺炎影响的实验研究

目的 探讨碘对小鼠产后甲状腺炎(postpartum thyroiditis,PPT)发生,发展的影响.方法 44只8周龄雌性C57BL/6J小鼠均饲以低碘饲料(含碘量≤35 μg/kg),按体质量随机分成4组:非妊娠实验性自身免疫性甲状腺炎(EAT)组 8只,采用猪甲状腺球蛋白加完全弗氏佐剂复制EAT模型,最终存活6只,NI-PPT组(正常碘),10HI-PPT组(10倍碘),50HI-PPT组(50倍碘)各12只,复制EAT模型(方法 同非妊娠EAT组)后,与性成熟雄鼠交配,分别有7,6,6只小鼠受孕.4 组鼠分别饮用含KI为0.3,0.3,3.0,15.0 mg/L的碘水.妊娠母鼠生产4周后,观察4组小鼠甲状腺组织病理改变,血清中甲状腺球蛋白抗体(Tg-Ab)和甲状腺过氧化物酶抗体(TPO-Ab)水平,血清中甲状腺激素(TT3,TT4)水平以及脾脏中IFN-γ,和IL-4 mRNA的表达水平.结果 甲状腺组织病理检查示甲状腺内炎细胞浸润,上皮细胞扁平,滤泡萎缩或破坏.非妊娠EAT组,NI-PPT组,10HI-PVT组小鼠的炎细胞浸润程度均低于50HI-PPT组,组间两两比较,差异有统计学意义(P＜0.05).非妊娠EAT组,NI-PPT组,10HI-PPT组和50HI-PPT组的血清TPO-Ab水平分别为(14.32±8.85)%,(64.45±10.52)%,(38.46±5.57)%,(90.09±9.98)%,任意两组组间比较,差异均有统计学意义(P＜0.05),血清Tg-Ab水平分别为(33.74±3.71)%,(29.65±2.06)%,(37.21±3.87)%,(33.87±4.17)%,组间比较差异无统计学意义(F＝0.484,P＞0.05),血清TT3水平分别为(2.47±0.69)%,(1.57±0.25)%,(1.60±0.28)%,(1.82±0.75)%,组间比较差异无统计学意义(F＝1.596,P＞0.05),非妊娠EAT组,NI-PPT组和10HI-PPT组血清中TT4水平[(99.87±5.97)%,(89.13±7.64)%,(91.05±5.82)%]与50HI-PPT组[(66.68±5.47)%]比较,差异均有统计学意义(P＜0.05).在非妊娠EAT组,NI-PPT组,10HI-PPT组和50HI-PPT组,小鼠脾脏IFN-γmRNA表达水平分别为1.02±0.10,1.37±0.10,1.39±0.12,1.68±0.06,除NI-PPT组和10HI-PVF组的组间比较之外,其他的任意两组组间比较.差异均有统计学意义(P＜0.05),IL-4 mRNA表达水平分别为0.24±0.05,0.35±0.05,0.49±0.04,0.53±0.06.10HI-PPT组,50HI-PPT组与非妊娠EAT组和NI-PPT组比较,差异均有统计学意义(P＜0.05).结论 妊娠及产后易诱发PPT,增加适量碘的摄入量,可降低PPT的炎症反应,而补碘过量也是诱发或增加产后甲状腺炎发生的重要因素.因此,妊娠及产后补碘应遵循合理和科学的原则.     

碘过量和多聚肌苷酸-聚胞苷酸及甲状腺球蛋白诱发小鼠甲状腺炎对Toll样受体3表达的影响

目的 观察碘过量(high iodine,HI)和多聚肌苷酸-聚胞苷酸[Polyinosinic-Polycytidylic acid,Poly (I:C),Poly]及甲状腺球蛋白(Thyroglobulin,TG)诱发小鼠甲状腺炎对Toll样受体3(Toll-like receptor 3,TLR3)表达的影响,探讨TLR3在自身免疫性甲状腺炎发病中的作用.方法 NOD(Non-obese diabetic)小鼠42只,体质量(20±3)g.按体质量将小鼠随机分为6组:对照组、HI组、Poly组、TG组、HI+TG组、HI+Poly组,每组7只.对照组:饮用去离子水,腹腔注射生理盐水0.1 ml,每天1次,连续1周,在处死小鼠前1周隔日1次,同样剂量生理盐水再注射3次;HI组:饮用0.05%的碘化钠去离子水,腹腔注射生理盐水(同对照组);Poly 组:饮用去离子水,腹腔注射0.1 ml Poly(1 g/L,按5 mg/kg体质量),每天1次,连续1周,处死前1周隔日1次,同剂量Poly再注射3次;TG组:饮去离子水,腹腔注射生理盐水(同对照组),皮下免疫猪TG 0.1 mg,在喂养第4、8周时分别再加强免疫1次,剂量减半;HI+Poly组:给药方法同HI组和Poly组;HI+TG组:给药方法同HI组和TG组.喂养8周后处死小鼠,取出甲状腺组织,冰冻切片、常规HE染色,光镜下观察小鼠甲状腺组织形态学变化:根据甲状腺组织炎细胞浸润数量及浸润范围、滤泡破坏范围等进行炎症程度分级;应用TLR3抗体对甲状腺切片进行免疫荧光染色,荧光显微镜下观察TLR3的表达,体视学分析甲状腺TLR3阳性细胞数密度变化.结果光镜下,Poly组甲状腺未见炎细胞浸润,HI组和TG组小鼠甲状腺都有不同程度的炎细胞浸润,HI+TG组和HI+Poly组甲状腺炎症细胞浸润和甲状腺滤泡破坏严重,炎症分级均在"++"以上.免疫荧光显示.HI组和Poly组的甲状腺滤泡上皮细胞可见到TLR3表达,在HI组和HI+Poly组炎症区域出现TLR3表达强阳性的炎症细胞.体视学分析甲状腺TLR3阳性细胞数密度,对照组、HI组、Poly组、TG组、HI+TG组、HI+Poly组组间比较差异有统计学意义(F=7.870,P<0.01);与对照组[(0.062±0.025)mm2]比较,HI+Poly组[(9.287±0.522)mm2]增加最为显著(P<0.01),而且HI+Poly组高于HI组[(2.570±0.257)mm2]和Poly组[(1.361±0.148)mm2,P均<0.01],HI+TG组[(4.843±0.405)mm2]高于HI组和TG组[(1.601±0.268)mm2,P均<0.01].结论 HI和TG免疫可诱发NOD鼠发生甲状腺炎,并刺激甲状腺滤泡上皮表达TLR3,Poly加重了HI诱发的NOD鼠甲状腺炎的病理变化过程;浸润的炎症细胞中亦有TLR3强阳性的细胞,提示TLR3途径参与了自身免疫性甲状腺炎的发病过程.     

不同碘水平对哺乳期母鼠和仔鼠碘代谢及甲状腺功能影响的研究

目的 研究不同碘水平对哺乳期母鼠和仔鼠的碘代谢及甲状腺功能的影响.方法 Wistar母鼠随机分为4组:重度缺碘组(SID),轻度缺碘组(MiID),正常碘组(NI),碘过量组(ExI).所有大鼠均食用缺碘饲料,饮水给予不同剂量的碘化钾,喂养3个月后交配,检测哺乳14 d时母鼠及其仔鼠的尿碘、乳汁碘、血碘和血液甲状腺激素(TH)水平,测定母鼠甲状腺重量,观察母鼠和仔鼠甲状腺形态学改变.结果 (1)母鼠及仔鼠尿碘、乳汁碘和血碘均随饮食碘供给量的增加而增加,其组间变化幅度以尿碘为最高、乳汁碘次之、血碘最低.(2)与NI组比较,SID组母鼠血清TT44降低[(16.7±12.0对36.4±15.0)nmol/L,P<0.05],TSH[(5.73+2.90对1.38+0.30)mIU/L,P<0.01]和TT3/TT4比值(6.6+2.7对2.1±0.3,P<0.01)升高,仔鼠TT4[(10.6+2.3对16.4±4.7)nmoL/L,P<0.05]降低;MiID和ExI组无论母鼠和仔鼠均与NI组无明显差异.(3)SID组母鼠和仔鼠发生典型小滤泡增生性甲状腺肿,MiID组母鼠呈现轻度甲状腺肿,ExI组母鼠甲状腺呈现轻度多形性特征,而两组仔鼠甲状腺与NI组无明显差别.结论 重度缺碘会导致母-子甲状腺功能减退,但在轻度缺碘和碘过量时通过母体和仔代的代偿作用可以维持母-子正常的碘营养和甲状腺功能.     

Thyroid iodine content and serum thyroglobulin: cues to the natural history of destruction-induced thyroiditis

Twenty-eight patients with destructive thyroiditis were followed to study the natural history of healing of thyroid gland injury. All had sequential measurements of thyroidal iodine [127I] content by fluorescent scanning (normal mean, 10.1 mg), 17 had serial serum thyroglobulin (Tg) measurements (normal, less than 21 ng/ml), and 13 had perchlorate discharge studies during the recovery phase. Seventeen patients had painful subacute thyroiditis (SAT), 9 had painless thyroiditis with thyrotoxicosis (PTT), and 2 had postpartum thyroiditis with thyrotoxicosis (PPT). Thyroidal iodine content decreased from a mean of 9.8 to a nadir of 3.8 mg in patients with SAT and from 8.5 to a nadir of 3.5 mg in patients with PTT. Mean serum Tg concentrations were highest (approximately 165 ng/ml) in both groups 1-3 months after the onset of symptoms. Abnormalities in both 127I content and Tg levels persisted for 2 or more yr in some individuals. No patient had detectable Tg antibodies by hemagglutination, but low titers were detected intermittently by sensitive RIA in 5 PTT patients. Microsomal antibodies were positive in only 1 of 16 SAT patients, but in 4 of 7 PTT patients and in both PPT patients. Three patients had positive perchlorate discharge tests (2 of 8 with SAT, 0 of 4 with PTT, and 1 of 1 with PPT). Permanent hypothyroidism occurred in 3 patients (2 with PTT; 1 with SAT and positive antibodies), but did not correlate with perchlorate results. HLA typing and serum immunoglobulin measurements were not useful for predicting the clinical course. These data indicate that several years may be necessary for complete resolution of destructive thyroiditis; many patients have evidence of thyroid injury persisting long after serum thyroid hormone and TSH levels become normal.     

Iodine and thyroid autoimmune disease in animal models.

Thyroid autoimmune diseases are complex, polygenic afflictions the penetrance of which is heavily dependent on various environmental influences. In their pathogenesis, an afferent stage (enhanced autoantigen presentation), a central stage (excessive expansion and maturation of autoreactive T and B cells), and an efferent stage (effects of autoreactive T cells and B cells on their targets) can be discerned. At each stage, a plethora of inborn, endogenous or exogenous factors is able to elicit the abnormalities characteristic of that stage, thus opening the gateway to thyroid autoimmunity. Iodine is an important exogenous modulating factor of the process. In general, iodine deficiency attenuates, while iodine excess accelerates autoimmune thyroiditis in autoimmune prone individuals. In nonautoimmune prone individuals, the effects of iodine are different. Here iodine deficiency precipitates a mild (physiological) form of thyroid autoimmune reactivity. Iodine excess stimulates thymus development. Iodine probably exerts these effects via interference in the various stages of the autoimmune process. In the afferent and efferent stage, iodine-induced alterations in thyrocyte metabolism and even necrosis most likely play a role. By contrast, in the central phase, iodine has direct effects on thymus development, the development and function of various immune cells (T cells, B cells macrophages and dendritic cells) and the antigenicity of thyroglobulin.