人小胶质细胞中促甲状腺激素受体的表达、分布及活性观察  

Expression,distribution,and activity of TSHR in human microglia

作　　者：陈雪兰 傅士恩 匡雅琪 朱莹丽 黄振兴[1] 杨海燕[1] 罗佐杰[1] CHEN Xuelan;FU Shien;KUANG Yaqi;ZHU Yingli;HUANG Zhenxing;YANG Haiyan;LUO Zuojie(Department of Endocrinology,The First Affiliated Hospital of Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西医科大学第一附属医院内分泌科,南宁530021

出　　处：《山东医药》2024年第14期18-21,共4页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(82260159)。

摘　　要：目的 探讨人小胶质细胞(HMC3)中促甲状腺激素受体(TSHR)的表达、分布及活性。方法 取人小胶质细胞HMC3,以人甲状腺细胞及肝细胞作为对照,采用逆转录聚合酶链式反应检测TSHRN mRNA表达,以验证TSHR mRNA转录物。通过Western blotting法和免疫荧光染色分析测定HMC3细胞中TSHR蛋白水平及分布。将HMC3细胞分为0、20、50、100、200 ng/mL促甲状腺激素受体刺激抗体(TSAb)组和阴性对照组,将各组细胞加入100μmol/L磷酸二酯酶抑制剂IBMX预孵育60 min,然后向TSAb组分别加入相应浓度的TSHR特异性抗体TSAb培养24 h。使用免疫测定试剂盒测定各组环磷酸腺苷(cAMP)水平,验证TSHR在HMC3细胞中的活性。结果 在HMC3细胞中检测到TSHR mRNA,其序列与人甲状腺细胞及肝细胞来源的mRNA相同。TSHR蛋白在HMC3细胞的细胞膜中表达。与0 ng/mL TSAb组相比,20 ng/mL TSAb组cAMP水平无统计学差异,50、100、200 ng/mL TSAb组cAMP水平增加,且高浓度组cAMP水平高于低浓度组(P<0.05或<0.01)。结论 TSHR在人小胶质细胞中存在并表达于细胞膜,且具有信号传导功能。Objective To investigate the expression,distribution,and activity of thyroid-stimulating hormone receptor(TSHR)in human microglia(HMC3).Methods In this study,we used human microglial cells(HMC3)as the experimental group and human thyroid cells and hepatocytes as the controls.Reverse transcription polymerase chain reaction was used to detect the expression of TSHR mRNA to identify the transcript of TSHR mRNA.Western blotting and immunofluorescence staining analysis was used to detect TSHR protein level and the distribution in HMC3 cells.HMC3 cells were divided into the 0,20,50,100,200 ng/mL thyroid-stimulating hormone receptor-stimulat⁃ing antibody(TSAb)groups and negative control group.Each group was preincubated with 100μmol/L phosphodies⁃terase inhibitor(IBMX)for 60 minutes.The corresponding concentrations of TSAb were added to cells of the TSAb groups and then they were incubated for 24 h.We measured cyclic adenosine monophosphate(cAMP)levels by immu⁃noassay kits to verify the functional activity of TSHR in HMC3 cells.Results TSHR mRNA was detected in HMC3 cells with the same sequence as mRNAs derived from human thyroid cells and hepatocytes.TSHR protein was ex⁃pressed in the cell membrane of HMC3 cells.Compared with the 0 ng/mL TSAb group,there was no significant statis⁃tical difference in cAMP levels in the 20 ng/mL TSAb group.However,cAMP levels increased in the 50,100,and 200 ng/mL TSAb groups,and the high-concentration group had higher cAMP levels than the low-concentration group(P<0.05 or P<0.01).Conclusion TSHR is expressed in the cell membrane of human microglia and has a signal⁃ing function.

关 键 词：促甲状腺激素受体 小胶质细胞 促甲状腺激素受体刺激抗体 环磷酸腺苷 

分 类 号：R581[医药卫生—内分泌]