摘要
目的探讨促甲状腺激素受体(TSHR)在人微血管内皮细胞(HMEC-1)的表达及其活性。方法体外培养HMEC-1,采用逆转录-聚合酶链反应(RT-PCR)法检测细胞TSHR mRNA的表达;将细胞接种在六孔板上,分空白对照组、低剂量组、中剂量组和高剂量组,空白对照组给予不含促甲状腺激素(TSH)的基础培养基培养,低剂量组、中剂量组和高剂量组分别给予0.10 U/L,1.00 U/L,10.00 U/L的TSH干预。24 h后采用酶联免疫吸附法(ELISA)检测细胞内环磷酸腺苷(c AMP)水平,研究HMEC-1细胞中的TSHR是否具有活性。显著性检验采用单因素方差分析,组间比较采用LSD-t法。结果 RT-PCR结果显示HMEC-1表达TSHR mRNA;ELISA结果显示TSH刺激HMEC-1后细胞内cAMP表达增加,低剂量组cAMP水平[(5.23±0.86)mmol/L]与空白对照组[(1.77±0.25)mmol/L]比较,差异有统计学意义(t=3.47,P<0.01),并且此效应呈剂量依赖性(F=92.36,P<0.01)。结论 HMEC-1细胞中存在TSHR,且该受体可与TSH特异性结合发挥作用。
Objective To investigate the expression and activity of thyroid stimulating hormone receptor(TSHR)in human microvascular endothelial cells(HMEC-1).Methods Culture HMEC-1 cells in vitro and detected TSHR mRNA by reverse transcription-polymerase chain reaction(RT-PCR).The cells were seeded on the six-well plate and divided into blank control group,low dose group,middle dose group and high dose group.The blank control group was cultured with basal medium without thyroid stimulating hormone(TSH).The low dose group,middle dose group and high dose group were given 0.10 U/L,1.00 U/L,10.00 U/L TSH intervention,respectively,and after 24 h,the intracellular cyclic adenosine monophosphate(cAMP)content was measured by enzyme-linked immunosorbent assay(ELISA)to study whether TSHR on HMEC-1 was active.Significant test was analysed by one-way anova,and LSD-t was used on amparion among groups.Results RT-PCR results showed that HMEC-1 expressed TSHR mRNA.ELISA results showed that the expression of cAMP increased in HMEC-1 stimulated by TSH,there was significant difference between low dose group and blank control group[(5.23±0.86)mmol/L,(1.77±0.25)mmol/L,t=3.47,P<0.01],and this effect was dose-dependent(F=92.36,P<0.01).Conclusion TSHR is present in HMEC-1,and the receptor may interact specifically with TSH.
作者
孟强
时敏敏
陈静
伊鹏飞
王娜
孙琳
Meng Qiang;Shi Minmin;Chen Jing;Yi Pengfei;Wang Na;Sun Lin(Department of Endocrionology,Affiliated Hospital of Jining Medical University,Jining 272029,China)
出处
《中华诊断学电子杂志》
2018年第1期69-72,共4页
Chinese Journal of Diagnostics(Electronic Edition)
基金
山东省自然科学基金(ZR2014HL031)
关键词
受体
促甲状腺素
微血管
内皮细胞
Receptors,thyrotropin
Microvessels
Endothelial cells
作者简介
通讯作者：孙琳，Email：sunlinsddx@sina.com
