TSH和TSAb刺激正常人甲状腺细胞cAMP生成的研究

以改良Hank's液溶解的bTSH和TSAb刺激体外培养的正常人甲状腺细胞。结果,甲状腺细胞释放cAMP 的量随细胞含量(5～20×10~4/孔)、bTSH 浓度(0～10~4mu/L)或病人血清用量(0～350μl/孔)的增加而增加,并且,当细胞培养时间(0～96h)或刺激时间(0～6h)延长时,cAMP 的生成量亦升高。提示体外培养的甲状腺细胞完整而具活力。本研究为临床检测TSAb 奠定了实验基础。     

肾上腺素能神经递质对人正常甲状腺细胞cAMP生成的影响

研究肾上腺素(E)、去甲肾上腺素(N)和异丙肾上腺素(1)对培养的人正常甲状腺细胞生成cAMP的影响。发现三种神经递质(10~(-3)～10~(-5)M)均可促进甲状腺细胞基础cAMP的释放,cAMP产值与E、N或Ⅰ间存在明显的时效(0～4小时)和量效关系。E或Ⅰ与TSH联用,可增强对甲状腺细胞的刺激效应,N与TSH合用,cAMP释放显著低于二者单独使用。提示甲状腺细胞受肾上腺素能神经递质兴奋和抑制的双重调节。     

正常人甲状腺单层细胞培养甲状腺球蛋白分泌的研究

用手术切除的甲状腺腺瘤旁相对正常的甲状腺组织10例,经组织制备与甲状腺单层细胞培养。用bTSH10、10~2和10~3μu/ml的三个浓度及先后分0天、3天和6天三个时间,观察不同浓度bTSH与不同培养天数对甲状腺单层细胞产生Tg值的变化,未用bTSH的基础培养下0天的Tg基础对照值为每孔4.90±0.07ng/ml,3天、6天升高不显著。各浓度bTSH刺激甲状腺细胞后第3天6天与0天,以及6天与3天相比均有显著升高。各浓度bTSH刺激后Tg的分泌值与相应日对照值均有显著升高,而各浓度间两两比较,仅第6天在bTSH三个浓度刺激下其释放Tg值有量效关系,而第3天则无统计学差别。实验结果证明正常人甲状腺单层细胞培养中bTSH具有促进分泌的能力。     

转染中国仓鼠卵巢细胞人TSH受体的功能评价

目的 研究转染中国仓鼠卵巢(CHO)细胞人促甲状腺激素(hTSH)受体(hTSHR)的功能.方法 抽提已转染hTSHR的CHO细胞基因组DNA,PCR扩增目的条带,并对产物进行序列测定.采用受体的放射配体结合分析法,以固定量125I的牛促甲状腺激素(125I-bTSH)和浓度递增的未标记bTSH与hTSHR发生竞争结合反应分析受体的基本功能;通过TSH刺激试验测定细胞环磷酸腺苷(cAMP)含量,评价hTSHR的生物学活性.结果 PCR扩增片段长度与预期相符,且序列完全正确.竞争结合反应表明,随着bTSH浓度的递增,125I-bTSH与hTSHR的结合量逐渐下降;TSH刺激试验显示,随着bTSH浓度的递增,细胞cAMP含量亦逐渐升高,当bTSH浓度为10 mIU/mL时,cAMP水平达到峰值.结论 cDNA序列整合在CHO细胞基因组内并在细胞膜上表达的hTSHR,具有受体的基本功能特性和良好的生物学活性.该实验为自身免疫性甲状腺疾病患者血清甲状腺刺激性抗体(TSAb)和阻断性抗体(TSBAb)的检测奠定了基础.     

转染中国仓鼠卵巢细胞人TSH受体的功能评价

目的研究转染中国仓鼠卵巢(CHO)细胞人促甲状腺激素(hTSH)受体(hTSHR)的功能。方法抽提已转染hTSHR的CHO细胞基因组DNA,PCR扩增目的条带,并对产物进行序列测定。采用受体的放射配体结合分析法,以固定量125I的牛促甲状腺激素(125I-bTSH)和浓度递增的未标记bTSH与hTSHR发生竞争结合反应分析受体的基本功能;通过TSH刺激试验测定细胞环磷酸腺苷(cAMP)含量,评价hTSHR的生物学活性。结果PCR扩增片段长度与预期相符,且序列完全正确。竞争结合反应表明,随着bTSH浓度的递增,125I-bTSH与hTSHR的结合量逐渐下降;TSH刺激试验显示,随着bTSH浓度的递增,细胞cAMP含量亦逐渐升高,当bTSH浓度为10 mIU/mL时,cAMP水平达到峰值。结论cDNA序列整合在CHO细胞基因组内并在细胞膜上表达的hTSHR,具有受体的基本功能特性和良好的生物学活性。该实验为自身免疫性甲状腺疾病患者血清甲状腺刺激性抗体(TSAb)和阻断性抗体(TSBAb)的检测奠定了基础。     

锂对甲状腺细胞内信号系统的影响

锂对猪甲状腺细胞的 TSH/Forskolin 刺激的 cAMP 累积有抑制作用,对 TSH 引起细胞内游离钙浓度升高反应亦有抑制作用。锂诱发的甲状腺机能减退可能是由于锂对甲状腺细胞内主要的信号系统(腺苷酸环化酶—cAMP 系统和钙信号系统)的抑制作用而引起的。     

甲状腺内注射地塞米松治疗甲状腺机能亢进80例观察

将８０例甲状腺机能亢进症（甲亢）患者随机分为两组，治疗组５０例，对照组３０例，两组患者均常规给予抗甲状腺药物治疗，待甲亢症状缓解，Ｔ３、Ｔ４正常或接近正常时，加用甲状腺片４０ｍｇ／ｄ。治疗组在常规治疗的基础上给予地塞米松５－１０ｍｇ加入２％普鲁卡因２ｍｌ，分别注入甲状腺两叶内或结节内，第１次，１０次为一疗程。结果治疗组甲状腺缩小显著，所需时间也较对照组短。     

MHC-Ⅱ类分子与hTSH受体共表达系统的建立

目的:应用电击基因导入法,建立MHC-Ⅱ类分子与hTSHR共表达体系(hMl2),为GD发病机制的研究奠定基础.方法:采用电击基因导入法将质粒pCRTM3-hTSHR转染于M12细胞(H-2d),应用G418筛选出阳性细胞,经有限稀释获得单克隆细胞株.用逆转录聚合酶链反应(RT-PCR)法检测单克隆细胞内hTSHR胞外段mRNA的表达;用甲亢病人血清粗提IgG及bTSH刺激后测定上清中cAMP值以检测单克隆细胞株表达的hTSHR是否具有天然hTSHR功能.结果:应用有限稀释法获得14株单克隆细胞.RT-PCR结果显示:3号、5号、6号、8号、11号细胞内有高水平的hTSHR胞外段mRNA的转录.应用甲亢病人血清粗提IgG及bTSH刺激后,6号、8号细胞上清中cAMP值明显升高,与对照组之间存在显著差别(P＜0.05).结论:应用电击基因导入法成功地建立了MHC-Ⅱ类分子与hTSH受体共表达体系.     

诱生的NO对原代培养人甲状腺细胞的影响

目的 研究原代培养人甲状腺细胞在IL-1α刺激下NO生成情况及NO对甲状腺细胞毒作用和对甲状腺功能影响。方法 IL-1α和L—NAME刺激甲状腺细胞48h，测定培养液中NO含量和LDH活力以及细胞中TPO活性；用SNP刺激甲状腺细胞4h，测定甲状腺细胞碘有机化。结果 IL-1α剂量依赖性诱导甲状腺细胞产生NO；IL-1α能提高甲状腺细胞LDH释放，降低TPO活性，L-NAME可抑制上述作用；SNP剂量依赖性抑制甲状腺细胞碘有机化。结论 IL-1α能诱导甲状腺细胞产生大量NO；NO对甲状腺细胞有部分细胞毒作用．可抑制TPO活性和甲状腺细胞碘有机化．     

用中国仓鼠卵巢细胞测定甲状腺刺激性抗体实验条件的探讨

目的探讨用中国仓鼠卵巢促甲状腺素受体细胞(Chinese hamster overy-thyrotropin receptor cell,CHO-TSHR细胞)测定甲状腺刺激性抗体(TSAb)的实验条件。方法参照文献方法,观察不同培养稀释液、细胞浓度、细胞培养时间、IgG浓度及IgG刺激时间对CHO-TSHR细胞cAMP生成的影响。结果无NaCl的Hanks液可提高cAMP的生成量;细胞浓度为1×105孔-1、细胞培养48 h、IgG浓度为1.0 g.L-1、IgG刺激时间为3 h时cAMP生成量最大。结论用CHO-hTSHR细胞作TSAb测定的合适条件为:(1)采用无NaCl的Hanks液做培养稀释液;(2)细胞浓度为1×105孔-1;(3)细胞培养时间为48 h;(4)IgG用量为1.0 g.孔-1;(5)IgG刺激时间为3 h。     

甲状腺细胞的简易冷冻保存方法

以含４０％小牛血清、８％二甲亚砜的Ｅａｇｌｅ培养液作为冻存液，采用温度梯度冷冻法保存人正常或Ｇｒａｖｅｓ病（ＧＤ）甲状腺细胞，结果显示，冷冻保存３、６或１２个月的甲状腺细胞成活率均大于８５％，各冻存时相的细胞对ＴＳＨ刺激生成ｃＡＭＰ的反应性无明显差异，并与非冷冻甲状腺细胞的反应强度相似。提示冻存一年内的甲状腺细胞仍保持良好的活力与功能，可以作为体外，研究甲状腺生理功能和病理变化的材料。     

人绒毛膜促性腺激素与甲状腺功能关系的实验室和临床研究

采用细胞培养方法研究孕妇甲状腺刺激活性（ＴＳＡｂ）及人绒毛膜促性腺激素（ｈＣＧ）对甲状腺细胞环磷酸腺苷（ｃＡＭＰ）生成的调节作用。结果发现：①早孕组（孕８￣１２周）及产前组（孕３４￣３８周）血清促甲状腺激素（ＴＳＨ）水平明显降低，早孕组ＴＳＨ与血ｈＣＧ显著负相关。②早孕组及产前组血清ＩｇＧ的ＴＳＡｂ明显高于对照组，尤以早孕组为著，两组孕妇ＴＳＡｂ阳性率分别为７９％和５６％，ＴＳＡｂ可显著刺激甲状腺     

The effect of thyrotropin receptor antibodies on the proliferation of FRTL-5 cells and the expression of protooncogene c-fos mRNA.

ＴｈｅｅｆｅｃｔｏｆｔｈｙｒｏｔｒｏｐｉｎｒｅｃｅｐｔｏｒａｎｔｉｂｏｄｉｅｓｏｎｔｈｅｐｒｏｌｉｆｅｒａｔｉｏｎｏｆＦＲＴＬ５ｃｅｌｓａｎｄｔｈｅｅｘｐｒｅｓｉｏｎｏｆｐｒｏｔｏｏｎｃｏｇｅｎｅｃｆｏｓｍＲＮＡＦａｎｇＰｅｉｈｕａ方佩华，ＬüＭ...     

低渗透压反应液促进甲状腺细胞合成cAMP机理

应用ＦＲＴＬ－５大鼠甲状腺细胞研究了无氯化钠低渗透压反应液促进促甲状腺激素（ＴＳＨ）刺激的甲状腺细胞合成环磷酸腺苷（ｃＡＭＰ）的机理。结果：无氯化钠低渗液组细胞内和细胞外液ｃＡＭＰ浓度均高于含氯化钠等渗液组。无氯化钠低渗液组细胞外液ｃＡＭＰ与总ｃＡＭＰ比率高于对照组。低渗液组细胞内和细胞外液ｃＡＭＰ浓度随反应时间延长持续升高；等渗组细胞内ｃＡＭＰ浓度在反应１０ｍｉｎ后达最高水平，以后无增加。结果表明，无氯化钠低渗反应液可能对提高甲状腺细胞膜ｃＡＭＰ的通透性，增强ＴＳＨ受体后反应有一定作用。     

正常和甲亢甲状腺细胞检测甲状腺刺激抗体的对比研究

采用细胞培养方法,对比研究正常和Graves病(GD)甲状腺细胞检测甲状腺刺激抗体(TSAb)的敏感性、特异性及重复性.结果显示:(1)正常和GD甲状腺细胞检测的TSAb活性具有显著相关性.(2)GD病人血清TSAb活性与甲状腺球蛋白抗体(TGAb)和甲状腺过氧化物酶抗体(TPOAb)无明显相关关系.(3)正常甲状腺检测GD组及桥本氏病(HT)组的阳性率分别为0.926和0.182,而GD甲状腺测定的阳性率为0.889和0.182.提示正常和GD甲状腺检测TSAb均具有特异性强、敏感性高和重复性好的特点,GD细胞测定TSAb有更高的实用价值.     

Thyroid status of normal pregnant women in Dhaka City

The thyroid gland secretes thyroid hormones having important role in embryogenesis and fetal development during pregnancy. The present study was carried out to find out alteration in the thyroid size and serum thyroid hormonal levels (T4, T3 and TSH) in normal pregnant women as compared to non-pregnant women in Dhaka City. For this purpose, thirty-five pregnant women during third trimester attending OPD Dhaka Medical College Hospital from January 1998 to July 1998 were selected randomly and twenty-one non-pregnant women of childbearing age group were selected as control. The study revealed a high prevalence of goiter in both pregnant (34.2%) and non-pregnant women (28.5%). The study showed that the serum TT4 level of non-pregnant women was normal (mean +/- SD = 113 +/- 23 nmol/L) and the serum TT4 of pregnant mother during third trimester was significantly higher (mean +/- SD = 200 +/- 63 nmol/L, P < 0.001) compared to that of non-pregnant women. The serum TT3, levels of both pregnant and non-pregnant women were within normal range without significant difference between them. It was found that the serum TSH level of pregnant mother was within normal limit and was closely similar to that of non-pregnant women showing no significant difference between them. It can be concluded from the study that although goiter was present in a considerable number of pregnant women but they were euthyroid status as observed by increased production of serum total thyroxin (TT4) by thyroid to fulfill the maternal requirement, with nearly unchanged TT3 and TSH secretion.     

Hyperthyroidism due to inappropriate TSH secretion with associated hyperprolactinaemia--a case report and review of the literature.

A patient with inappropriate thyrotrophin (TSH) secretion is described. She initially presented with classical hyperthyroidism during pregnancy, responded to propylthiouracil and, subsequently, had a normal delivery. Hyperthyroidism persisted and 7.5 months later a subtotal thyroidectomy was performed. After a further 16 months, mild symptoms of hyperthyroidism recurred. She again responded to propylthiouracil, but developed galactorrhoea. At that stage, it was noted that she had persistently elevated circulating TSH in the presence of elevated T4 and T3 levels. Her symptomatology was mild, although objective indices of thyroid activity, including pulse rate, BMR, sex hormone binding globulin and cholesterol, were indicative of hyperthyroidism. CT scan and tomography of the sella were normal. She had a markedly exaggerated TSH response to thyrotrophin releasing hormone (TRH). Basal TSH and responsiveness to TRH was suppressed by high dose dexamethasone. The TSH response to TRH was partially suppressed by exogenous T3, but there was no effect on basal TSH levels. TSH also decreased slightly with L-dopa and bromocriptine. Circulating TSH rose markedly during methimazole administration. TSH alpha and beta subunits were elevated and appropriate for the high TSH. In addition, both subunits increased following TRH. The patient had basal hyperprolactinaemia with an impaired prolactin (PRL) response to TRH and metoclopramide. PRL suppressed with L-dopa and bromocriptine. The remaining anterior pituitary function was intact. Most of the laboratory findings argue against the presence of a TSH producing pituitary tumour and the most likely cause for inappropriate TSH secretion in this patient is selective resistance of the thyrotroph to thyroid hormones. A mild element of peripheral resistance might also be present. The hyperprolactinaemia could be related to lactotroph resistance to thyroid hormone. The complexities of treatment in this patient are stressed. Therapy was initially attempted with low dose dexamethasone, but this had no effect. T3 treatment produced an exacerbation of her symptomatology and did not influence basal TSH, thyroid hormones, or 131I uptake. Bromocriptine administration for 11 months partially suppressed basal TSH without influencing T3 and there was an increase in T4. Methimazole did decrease her T4 and T3, but TSH and PRL rose to even greater levels. Her hyperthyroidism was eventually controlled with an ablative dose of 131I. Thyroid hormone will be given in an attempt to suppress her TSH.     

甲状腺球蛋白负反馈调节机制

甲状腺球蛋白（Tg）是甲状腺滤泡中最重要含量最丰富的蛋白质,临床作为甲状腺癌复发和持续状态的肿瘤标志物被广泛 深入的研究。传统观点认为Tg是甲状腺激素合成的物质基础,不参与调节甲状腺激素合成分泌,甲状腺激素合成分泌受甲状 腺-下丘脑-脑垂体轴负反馈调节机制调控。本文阐述的甲状腺球蛋白负反馈调节机制基于实验研究结论认为：甲状腺球蛋白对 甲状腺滤泡细胞合成甲状腺激素功能存在负向调控作用,可以拮抗促甲状腺激素TSH的正向调控作用,推测甲状腺滤泡细胞功 能是Tg和TSH共同作用的结果,并提出滤泡周期模型揭示甲状腺滤泡异质性的原因。本文综述甲状腺球蛋白负反馈调节机制 研究进展,展望了该研究的临床意义,对比甲状腺-下丘脑-脑垂体轴负反馈调节机制,说明两个理论之间的差异,加深对甲状腺 两个负反馈调节机制的认识。