人促甲状腺激素受体的制备及其敏感性测定  

Preparation and Sensitivity Measure of Human Thyrotropin (TSH) Receptor

作　　者：贺铭[1] 杨永年[1] 

机构地区：[1]上海医科大学中山医院内分泌科

出　　处：《上海铁道大学学报》1999年第3期22-24,共3页

摘　　要：目的探索一项简便、经济的提取人甲状腺促甲状腺激素（ｔｈｙｒｏｉｔｒｏｐｉｎ，ＴＳＨ）受体－Ｇｓ蛋白－腺苷酸环化酶信息传递系统及检测ＴＳＨ受体敏感性的方法。方法用高速离心法，从低温保存的人甲状腺组织提取细胞膜蛋白ＴＳＨ受体及与之偶联的Ｇｓ蛋白－腺苷酸环化酶，以腺苷酸环化酶的活性反映ＴＳＨ受体与配基ＴＳＨ结合的敏感性。结果提取膜蛋白量（３０～１８０μｇ）与ｃＡＭＰ生成量之间呈显著的线性相关（ｒ＝０．９８７，Ｐ＜０．００５）；膜蛋白量相同时（５０μｇ），配基人ＴＳＨ（ｈＴＳＨ）浓度（０～２５００μＩＵ／ｍｌ）与ｃＡＭＰ生成量之间呈极显著的线性相关（ｒ＝０．９５４，Ｐ＜０．００１）；Ｇｓ蛋白兴奋剂ＧＴＰγＳ浓度（０～１．０ｍｍｏｌ／Ｌ）与ｃＡＭＰ生成量之间亦呈极显著性相关（ｒ＝０．９８６，Ｐ＜０．００１）。结论该方法制备到完整的ＴＳＨ受体－Ｇｓ蛋白－腺苷酸环化酶信号传递系统，腺苷酸环化酶活性能较好反映ＴＳＨ受体对配基ｈＴＳＨ刺激的敏感性。此方法简便经济，可用于甲状腺ＴＳＨ受体－Ｇｓ蛋白－腺苷酸环化酶信号传递系统的实验室研究。Objective To explore a simple and economical experimental method that can extract human thyroid TSH receptor G s protein Adenylate cycles (AC) signal pathway and measure the sensitivity of TSH receptor.Methods TSH receptor in cell membraone proteins and G s protein AC that coupled with TSH receptor was extracted from human thyroid preserved in low temperature by highspeed centrifugation.The sensitivity of TSH receptor combined with ligand human TSH (hTSH) was presented as the activity of AC.Results The amount of membrane protein preparation (30￣180 μg) was significantly related to the amount of cAMP produced ( r =0.987, P <0.05), the concentration of ligand hTSH (0￣2 500 μ IU/ml) and G s protein stimulator GTP γ S (0￣1.0 mmol/L) were highly significantly related to the amount of cAMP produced ( r =0.954, P <0.01 and r =0.986, P <0.001).Conclusion The perfect TSH receptor G s protein AC signal pathway was obtained by this method, and the activity of AC can perfectly stand for the sensitivity of TSH receptor combined with ligand hTSH, this simple and economic method can be used in experimental study in thyroid TSH receptor G s protein AC signal pathway.

关 键 词：甲状腺 受体 促甲状腺激素 G蛋白 敏感性 

分 类 号：R581[医药卫生—内分泌] R392－33[医药卫生—内科学]