Sun Feng-yuan, Pan Ye, Song Guo-xiang, Chang Ji-wu
Department of Ophthalmology, Second Hospital of Tianjin Medical University, Tianjin 300211, China.
Zhonghua Yan Ke Za Zhi. 2006 Feb;42(2):155-8.
To investigate gene and protein expression level of thyrotropin-receptor (TSHR) in retro-bulbar connective tissue and fat tissue in patients with thyroid-associated ophthalmopathy (TAO).
Retro-bulbar tissues specimens were obtained from 11 cases of TAO patients and 10 control cases during the operation. RNA was extracted from the tissue specimens using single-step method of acid guanidinium-thiocyanate-phenol-chloroform extraction and TSHR mRNA was analyzed by reverse transcription polymerase chain reaction (RT-PCR). Immunochemistry staining was performed in patients expressed TSHR, using a mouse polyclonal anti-TSHR peptide antibody to observe protein expression level of TSHR.
The size of TSHR mRNA segment is 521 bp. The rate of TSHR mRNA expression in TAO retro-bulbar tissue was 91% and was 20% in the control cases, the former was significantly greater than the later. Protein expression rate was 66.7% in TAO patients, and was negative in the controls.
High TSHR mRNA expression level and protein level in TAO retro-bulbar tissue and no expression in control tissue indicate that TSHR exists as a common antigen in the orbit and thyroid, and plays a key role in the pathogenesis of TAO.
探讨促甲状腺激素受体（TSHR）在甲状腺相关眼病（TAO）患者球后结缔组织和脂肪组织中的基因及蛋白表达水平。
手术中获取11例TAO患者及10例对照者的球后组织标本。采用酸胍-硫氰酸盐-苯酚-氯仿一步法从组织标本中提取RNA，通过逆转录聚合酶链反应（RT-PCR）分析TSHR mRNA。对表达TSHR的患者进行免疫化学染色，使用小鼠多克隆抗TSHR肽抗体观察TSHR蛋白表达水平。
TSHR mRNA片段大小为521 bp。TAO球后组织中TSHR mRNA表达率为91%，对照者为20%，前者显著高于后者。TAO患者蛋白表达率为66.7%，对照者为阴性。
TAO球后组织中TSHR mRNA表达水平及蛋白水平较高，而对照组织中无表达，表明TSHR作为眼眶和甲状腺中的共同抗原，在TAO发病机制中起关键作用。
