促甲状腺激素受体可在微血管内皮细胞表达∗

摘要

目的：：检测促甲状腺激素受体(TSHR)在小鼠脑微血管内皮细胞(bEnd.3)的表达。方法：取对数生长期 bEnd.3细胞,采用 RT-PCR 和 Western Boltting 方法检测 bEnd.3细胞 TSHR mRNA 和蛋白表达水平。结果：bEnd.3细胞总 RNA 浓度为2.9186μg/μl,A260/A280为1.97。TSHR mRNA 的扩增效率为96.41%±0.82%,溶解曲线呈单峰,相对表达量为0.64±0.14(相对睾丸组织)。电泳结果显示 bEnd.3细胞能清晰表达 TSHR mRNA。Western Blotting 显示 bEnd.3细胞可以检测到 TSHR 的α和β亚基蛋白表达,β亚基相对表达量为0.46±0.05(相对β-actin)。结论：小鼠脑微血管内皮细胞可表达 TSHR,该结果有助于临床研究 TSH 对血管内皮细胞功能的影响。%Objective:To evaluate the expression of thyroid stimulating hormone receptor (TSHR)on mouse microvascular endothelial cells (bEnd.3 ).Method:Real-time PCR and Western-boltting were used to analyze the expression of TSHR mRNA and TSHR protein in the bEnd.3 cells.Results:TSHR mRNA expression was detected in the bEnd.3 cells.The content of RNA in the bEnd.3 cells was 2.9186μg/μl,and its ratio of A260/A280 was 1. 97.TSHR was amplified and the amplification efficiency was 96.41%±0.82%.Dissociation curve from real-time PCR was unimodal.The relative quantity of TSHR mRNA in the bEnd.3 cells was 0.64±0.14,when the quantity of TSHR mRNA in the testicle was set as 1.0.TSHRαandβsubunit were detected by Western-blotting.The pro-tein ratio of TSHRβ/β-actin was 0.46±0.05 in the gray scale analysis.Conclusion:The expression of TSHR could be found in the bEnd.3 cells.This finding may be helpful to explore the effect of thyroid stimulating hormone on mi-crovascular endothelial cells.