促甲状腺激素受体,甲状腺过氧化物酶及甲状腺球蛋白基因表达…

为探讨促甲状腺激素受体。甲状腺过氧化物酶及甲状腺球蛋白ｍＲＮＡ在自身免疫甲状腺疾病（ＡＩＴＤ）病人甲状腺组织的表达情况及其相互关系，应用Ｎｏｒｔｈｅｒｎ印记杂交技术观察了ＴＳＨ受体、ＴＰＯ以及ＴＧ ｍＲＮＡ在７例Ｇｒａｖｅｓ病（ＧＤ），２例桥本甲状腺素炎（ＨＴ）病人甲状腺组织中的表达。研究显示：ＴＳＨ受体ｍＲＮＡ与ＴＰＯ和ＴＧ ｍＲＮＡ在ＧＤ组和ＨＴ组表达不一致，ＧＤ组ＴＳＨ受体ｍＲＮＡ与ＴＰＯ和     

桥本甲状腺炎和Graves病人类白细胞抗原—DR的免疫组化研究

为探讨甲状腺上皮细胞异常表达人类白细胞抗原（ＨＬＡ）－ＤＲ（ＤＲ抗原）与甲状腺自身免疫反应的关系，应用卵白素生物素复合物免疫组化法对１６例桥本甲状腺炎（ＨＴ）、１４例Ｇｒａｖｅｓ病（ＧＤ）和６例正常人ＤＲ抗原阳性甲状腺上皮细胞进行定位和定量研究，并与血清甲状腺球蛋白抗体（ＴＧＡｂ）、甲状腺过氧化物酶抗体（ＴＰＯＡｂ）值进行相关分析。结果显示，正常甲状腺上皮细胞不表达ＤＲ抗原。在所有被检ＨＴ和ＧＤ患者，均可见表达ＤＲ抗原的甲状腺上皮细胞，以ＨＴ组尤为多见。其主要分布于淋巴细胞浸润区，表达程度与血清ＴＧＡｂ、ＴＰＯＡｂ值密切正相关。提示浸润淋巴细胞可能促使甲状腺上皮细胞表达ＤＲ抗原，ＨＴ和ＧＤ患者血中出现ＴＧＡｂ、ＴＰＯＡｂ与其甲状腺上皮细胞异常表达ＤＲ抗原，易于受到免疫炎性损伤有关。     

自身免疫性甲状腺疾病树突状细胞和 HLA-DR 抗原的免疫组化研究

用ＡＢＣ免疫组化法研究桥本甲状腺炎（ＨＴ）和Ｇｒａｖｅｓ病（ＧＤ）患者甲状腺内的Ｓ１００蛋白阳性树突状细胞（ＤＣ）和ＨＬＡＤＲ（ＤＲ）抗原阳性甲状腺上皮细胞（ＴＥＣ）。发现ＨＴ和ＧＤ甲状腺内的Ｓ１００蛋白阳性ＤＣ较正常明显增多，且大多数ＤＣ与ＴＥＣ或浸润淋巴细胞密切接触。ＨＴ和ＧＤ的ＴＥＣ常见ＤＲ抗原异常表达，其主要分布在淋巴细胞浸润区。这说明ＤＣ异常增多、ＴＥＣ异常表达ＤＲ抗原可能与甲状腺自身免疫反应的启动和（或）延续有密切关系。     

甲状腺抗原诱导大鼠实验性自身免疫性甲状腺炎的研究

目的观察人类甲状腺球蛋白（ｈＴＧ），人类甲状腺过氧化物酶（ｈＴＰＯ）以及豚鼠促甲状腺激素受体（ｇＴＳＨＲ）免疫诱导Ｗｉｓｔａｒ大鼠产生实验性自身免疫性甲状腺炎（ＥＡＴ）的过程。方法雄性Ｗｉｓｔａｒ大鼠共分四组，每组１０只，三组分别多次注射ｈＴＧ、ｈＴＰＯ、ｇＴＳＨＲ，第四组注射生理盐水作为对照。自０至１９１天，每次注射后，分别测定循环中ＴＴ３、ＴＴ４水平以及ＴＧＡｂ、ＴＰＯＡｂ、ＴＲＡｂ、ＴＳＡｂ和ＴＢＡｂ的活性；ＥＡＴ的严重程度采用甲状腺切片的显微镜观察进行评估。结果与正常组比较，实验组大鼠血清中存在高水平的ＴＧＡｂ、ＴＰＯＡｂ和ＴＳＡｂ；且甲状腺组织可见不同程度的淋巴细胞浸润，类似人类桥本氏甲状腺炎。此外，在ＴＳＨＲ组中不仅血清ＴＳＡｂ显著升高，而且甲状腺组织显示滤泡上皮增生、乳头状突起等Ｇｒａｖｅｓ’病（ＧＤ）的病理学特征。结论三种异源性抗原均能诱发正常Ｗｉｓｔａｒ大鼠产生ＥＡＴ，ＴＳＨＲ免疫的大鼠还显示了ＧＤ的倾向。     

自身免疫性甲状腺疾病患者甲状腺过氧化物酶抗体测定的临床意义

我们对 176例自身免疫性甲状腺疾病 (ＡＩＴＤ)、5例亚急性甲状腺炎和 5 0例正常人做甲状腺过氧化物酶抗体 (ＴＰＯＡＢ)和甲状腺微粒体抗体 (ＴＭＡＢ)测定 ,并将二者进行比较。对象与方法1.对象 :ＡＩＴＤ患者 176例 ,男性 3 4例 ,女性 14 2例 ,年龄11～ 74岁 ,平均年龄 44 .5岁。 176例ＡＩＴＤ患者分为下列 4组 :(1)初诊Ｇｒａｖｅ’ｓ病组 (Ａ组 ) :3 9例 ,均未经抗甲状腺药物(ＡＴＤ)治疗 ,有高代谢临床表现 ,ＦＴ3 、ＦＴ4增高 ,ｓＴＳＨ减低 ,ＴＳＨ受体抗体 (ＴＲＡＢ)阳性。 (2 )Ｇｒａｖｅ’ｓ病ＡＴＤ治疗后甲亢已控制组 (Ｂ组 …     

甲状腺疾病患者血清TSAb和TBII的检测

采用细胞培养生物学方法和放射受体分析法检测甲状腺疾病患者血清甲状腺刺激抗体（ＴＳＡｂ）和促甲状腺激素结合抑制免疫球蛋白（ＴＢＩＩ）。发现Ｇｒａｖｅｓ病未治组ＴＳＡｂ和ＴＢＩＩ活性与阳性率明显高于Ｇｒａｖｅｓ病缓解组和桥本甲状腺炎（ＨＴ）组，后两组ＴｓＡｂ和ＴＢＩＩ活性与阳性率高于甲状腺腺瘤组和正常对照组。Ｇｒａｖｅｓ病未治组、缓解组和ＨＴ组其ＴＳＡｂ和ＴＢＩＩ的检出率分别为９１．７％和７９．２％、４８．０％和４４．０％及２３．１％和６１．５％。甲状腺腺瘤组和对照组ＴＳＡｂ和ＴＢＩＩ均为阴性，其活性亦无统计学差异。血清ＴＳＡｂ与ＴＢＩＩ活性以及ＴＳＡｂ、ＴＢＩＩ活性与Ｔ＿３、Ｔ＿４、ＦＴ＿３和ＦＴ＿４浓度之间均无显著相关性。提示ＴＳＨ受体抗体具有异质性，联合检测这两种抗体对了解自身免疫性甲状腺疾病（ＡＩＴＤ）的发病机理，合理诊治甲状腺疾病具有重要的临床价值。     

老年甲亢患者甲状腺刺激抗体检测及临床应用

以血清ＩｇＧ刺激培养的甲状腺机能亢进症患者甲状腺细胞，通过测定ｃＡＭＰ的释放值确定甲状腺刺激抗体（ＴＳＡｂ）的活性。结果显示：①老年Ｃｒａｖｅｓ病（ＧＤ）未治组ＴＳＡｂ活性显著高于ＧＤ缓解组与缓解停药组，３组患者ＴＳＡｂ的阳性率分别为８８．９％、４７．８％和４５．５％，甲状腺腺瘤组和对照组ＴＳＡｂ均为阴性。②ＧＤ缓解停药时５例ＴＳＡｂ阳性者中４例（８０．０％）于１年内复发，而６例阴性者仅１例（１６．７％）复发。③ＴＳＡｂ活性与血清Ｔ３、Ｔ４浓度无明显相关关系。提示ＴＳＡｂ测定对老年ＧＤ的诊断、鉴别诊断、治疗和预后判断有重要的指导意义。     

甲状腺功能低下大鼠中枢5-HT对TSH分泌的影响

目的 探讨甲状腺功能低下（甲低）大鼠中枢５－羟色胺（５－ＨＴ）系统的代谢变化与机制,以及５－ＨＴ对促甲状腺激素（ＴＳＨ）分泌的调节作用。方法 用０．５％ＮａＣＩＯ４诱导实验大鼠甲低功,采用荧光分析方法测定大鼠脑组织５－ＨＴ、５－ＨＩＡＡ含量及ＭＡＯ活性,放免法测定血清Ｔ３、Ｔ４及ＴＳＨ含量。结果 甲低组大鼠血清Ｔ３、Ｔ４含量明显下降,而血清ＴＳＨ含量及脑组织中５－ＨＩＡＡ、５－ＨＩＡＡ／５－ＨＴ比     

加碘盐和自身抗体对Graves病抗甲状腺药物治疗的影响

为探讨加碘盐 (ＡＩＳ)饮食和甲状腺自身抗体对Ｇｒａｖｅｓ病 (ＧＤ)病人抗甲状腺药物 (ＡＴＤ)治疗的影响 ,我们对 2 76例ＧＤ病人进行了平均 5年追踪研究 ,报告如下。一、资料和方法表 1　ＡＩＳ对ＧＤ病人ＡＴＤ治疗的影响 ( ｘ±ｓ)组别例数 治疗前甲状腺重量 (ｇ)首次缓解需时(ｄ)1年复发率(% )5年复发率(% )维持ＡＴＤ日用量(片 )缓解期ＦＴ3(ｎｇ/Ｌ)缓解期ＦＴ4 (ｎｇ/Ｌ)缓解期ＴＳＨ(ＩＵ/Ｌ)缓解期ＴＧＡｂ阳性率(% )缓解期ＴＰＯＡｂ阳性率(% )缓解期甲状腺重量 (ｇ)ＮＩＳ组 13 82 8.43± 45 .3 1± 1.45 3 7.681.68± 2 .43 …     

自身免疫性甲状腺病甲状腺浸润细胞亚群的观察

用免疫组织化学方法及单克隆抗体ＣＤ４、ＣＤ８、ＣＤ３、Ｉ２、Ｔａｃ对３０例ＧＤ患者和２２例ＨＴ患者甲状腺内浸润淋巴细胞进行观察。结果表明ＧＤ或ＨＴ的甲状腺内ＣＤ＋４Ｔ细胞数量均高于ＣＤ８＋Ｔ细胞；ＨＴ甲状腺内ＣＤ８＋Ｔ细胞、Ｉ２＋细胞、Ｔａｃ＋细胞百分数量显著高于ＧＤ甲状腺内相应细胞的百分数量。活化的淋巴细胞与血中ＴＧＡ和ＭＣＡ水平呈显著正相关。结合ＧＤ与ＨＴ的病理改变，提示ＨＴ甲状腺内免疫反应较ＧＤ时更为激烈，ＣＤ８＋Ｔ细胞在ＨＴ中的作用可能更为重要，甲状腺活化的淋巴细胞与自身抗体产生具有密切关系。     

Heterogeneity of TSH receptor-binding antibodies in Hashimoto's thyroiditis and Graves' disease

The possible heterogeneity of TSH receptor antibodies in Graves' disease (GD) and Hashimoto's thyroiditis (HT) with respect to the binding site on the receptor and corresponding biological effect was studied. Employing an immunoprecipitation assay (IPA), the sera of 80% of the patients with GD (24 out of 30) and 76% of the patients with HT (16 out of 21) contained TSH receptor-binding antibodies, compared to none of the sera from 17 normal volunteers and 8 patients with nontoxic multinodular goiter. TSH inhibited immunoprecipitation by GD and HT sera. In HT sera (n = 9), but not in GD sera (n = 5), heterogeneity of the TSH-induced inhibition was observed. Four HT sera showed complete inhibition of immunoprecipitation at a saturating concentration (19.8 nM) of TSH. Five HT sera, like the 5 GD sera, showed partial inhibition of immunoprecipitation by 19.8 nM TSH. Thyroid stimulating immunoglobulins (TSI) were found in four of the five GD sera and in only one of the nine HT sera. The results suggest that different subpopulations of TSH receptor antibodies, characterized by other receptor binding sites or different affinities, are associated with autoimmune thyroid disease.     

Autoimmunity involving the human sodium/iodide symporter: fact or fiction?

Presence, functional activity and clinical relevance of autoantibodies directed against the human sodium iodide symporter (NIS) in thyroid autoimmune diseases have become the subject of much controversy in recent years. Earlier reports have claimed that NIS may represent a major thyroid autoantigen that elicits formation of functionally relevant autoantibodies in a significant proportion of patients with Graves' disease (GD) and Hashimoto's thyroiditis (HT). Moreover, a recent study has extended this notion by reporting detection of NIS-autoantibodies in 22% and 24% of a small number of patients with GD and HT, respectively, but not in patients with other autoimmune diseases. However, in striking contrast to these reports, two independent groups of investigators have now presented convincing evidence that NIS-directed autoantibodies occur with low frequency among a large sample of patients with autoimmune thyroid diseases. Moreover, no evidence of specific iodide uptake inhibiting activity was obtained once sera had been subjected to dialysis and/or IgG extraction. Thus, although the controversy has not been definitively resolved, hNIS does not appear to be a major functionally relevant antigen in autoimmune thyroid diseases. Moreover, when detected in addition to TPO and TSH receptor autoantibodies, NIS-directed autoantibodies do not appear to contribute any diagnostic power for GD and HT.     

缝隙连接蛋白43、26在自身免疫性甲状腺疾病患者甲状腺上皮细胞中的表达

目的 探讨缝隙连接蛋白43、26(Cx43、Cx26)在自身免疫性甲状腺疾病(AITD)患者甲状腺上皮细胞中的表达,探索其在AITD的发病机制、转归及预后中所起的作用.方法 采用二步法免疫组织化学技术(S-P法)测定甲状腺腺瘤旁正常甲状腺组织(对照组,16例)、Graves病(GD组,30例)和桥本甲状腺炎(HT组,30例)患者甲状腺上皮细胞中Cx43、Cx26的分布及表达情况.结果 ①对照、GD和HT组甲状腺上皮细胞均有Cx43、Cx26的表达.既定位于细胞质中,又定位于细胞膜上.②对照、GD和HT组甲状腺上皮细胞Cx43表达阳性率分别为75.00%(12/16)、100.00%(30/30)和33.33%(10/30),Cx26表达阳性率分别为68.75%(11/16)、100.00%(30/30)和20.00%(6/30).Cx43、Cx26在GD组的表达强度均明显高于对照组(Z值分别为4.782、5.310,P均<0.017),在HT组的表达强度均明显低于对照组(Z值分别为2.703、3.123,P均<0.017).结论 Cx43、Cx26可在人甲状腺上皮细胞中表达;Cx43、Cx26在AITD的表达强度存在异质性,这种缝隙连接蛋白表达的异质性可能与AITD的发生、发展及预后有关.     

自身免疫性甲状腺疾病甲状腺组织中bcl-2家族蛋白的表达

目的研究凋亡相关基因bcl鄄2家族蛋白bcl鄄2、mcl鄄1、bcl鄄XL和bax在自身免疫性甲状腺疾病(AITD)甲状腺组织中的表达特征及与AITD发病机制之间的内在联系。方法以甲状腺腺瘤旁正常甲状腺组织为对照(C组,20例),采用免疫组织化学ElivisionTM二步染色法检测凋亡相关蛋白bcl鄄2、mcl鄄1、bcl鄄XL和bax在桥本甲状腺炎(HT组,33例)和Graves病(GD组,28例)患者甲状腺组织中的表达与分布。结果bcl鄄2蛋白表达强度GD组>C组>HT组(P<0.01);mcl鄄1蛋白表达强度GD组>C组>HT组(P<0.01);bcl鄄XL蛋白表达强度HT组和GD组强于C组(P<0.01),但HT组和GD组间差异无统计学意义(P>0.05);bax蛋白的表达强度HT组>GD组和C组(P<0.01),但GD组和C组间差异无统计学意义(P>0.05);HT组中,在淋巴细胞浸润区域附近的甲状腺滤泡上皮细胞(TEC)bcl鄄2表达弱,bax和mcl鄄1表达强;远离淋巴细胞浸润区域的TECbcl鄄2表达强,bax和mcl鄄1表达弱(P<0.05)。结论(1)抗凋亡bcl鄄2和mcl鄄1蛋白在HT中表达的减弱以及在GD中表达的增强对于HT甲状腺滤泡细胞凋亡的增加和GD甲状腺滤泡细胞的增殖可能起一定作用;(2)bax蛋白在HT中表达增强所起的促凋亡的作用对疾病的发生发展起一定作用;(3)bcl鄄2与bax表达强度的比值对于凋亡的调控起重要作用;(4)bcl鄄2家族蛋白bc     

Differential regulation of Fas-mediated apoptosis in both thyrocyte and lymphocyte cellular compartments correlates with opposite phenotypic manifestations of autoimmune thyroid disease.

Several mechanisms are probably involved in determining the evolution of autoimmune thyroid disease (AITD) towards either hypothyroidism and the clinical syndrome known as Hashimoto's thyroiditis (HT) or toward hyperthyroidism and the symptoms of Graves' disease (GD). To gain further insight into such mechanisms we performed an exhaustive comparative analysis of the expression of key molecules regulating cell death (Fas, Fas ligand [FasL], Bcl-2) and apoptosis in both thyrocytes and thyroid infiltrating lymphocytes (TILs) from patients with either GD or HT. GD thyrocytes expressed less Fas/FasL than HT thyrocytes, whereas GD TILs had higher levels of Fas/FasL than HT TILs. GD thyrocytes expressed increased levels of the antiapoptotic molecule Bcl-2 compared to the low levels detected in HT thyrocytes. The opposite pattern was observed in GD (low Bcl-2) and HT (high Bcl-2) TILs. The patterns of apoptosis observed were consistent with the regulation of Fas, FasL, and Bcl-2 described above. Our findings suggest that in GD thyroid the regulation of Fas/FasL/Bcl2 favors apoptosis of infiltrating lymphocytes, possibly limiting their autoreactive potential and impairing their ability to mediate tissue damage. Moreover, the reduced levels of Fas/FasL and increased levels of Bcl-2 should favor thyrocyte survival and favor the thyrocyte hypertrophy associated with immunoglobulins stimulating the thyrotropin (TSH) receptor. In contrast, the regulation of Fas/FasL/Bcl2 expression in HT promotes thyrocyte apoptosis, tissue damage, and a gradual reduction in thyrocyte numbers leading to hypothyroidism. These findings help define key molecular mechanisms contributing to the clinical outcome of thyroid autoimmunity.     

Difference in species specificity of TSH receptor antibodies in Graves' disease and Hashimoto's thyroiditis

TSH receptor antibodies have been detected in the sera of patients with Graves' disease (GD) and Hashimoto's thyroiditis (HT). Since non-human thyroid tissue fractions or cells are used in the majority of assays, the species specificity of TSH receptor antibodies in GD or HT could be important. The species specificity was evaluated by means of an immunoprecipitation assay (IPA) using Triton X-100 solubilized TSH receptors prepared from human, porcine and rat thyroid as well as guinea pig fat cells (GPFC). In each assay the majority (4 or 5) of the 6 patients with GD were IPA-positive. In contrast, 9 out of 11 patients (82%) with HT had a positive human and rat IPA, while only 3 out of 11 (27% p less than 0.05) sera were positive in the porcine and GPFC assays. Conclusions: no species specificity of TSH receptor antibodies was detected for patients with GD; a selective species specificity for human and rat TSH receptors was found for HT sera. This suggests that TSH receptor antibodies in HT either recognize different determinants on the receptor than the antibodies in GD or are of lower affinity. Furthermore, the use of porcine thyroid tissue or GPFC may lead to an underestimation of the presence and level of TSH receptor antibodies in patients with HT.