VR1+细胞模型的构建和Nielsen假说的验证

为研究类香草素受体(vanilloid receptortype 1,VR1)作为刺激性室内空气污染物的触发性生物靶分子的生理作用,在VR1-cDNA质粒基础上,应用分子克隆和细胞转染技术构建了体外培养的VR1 -HEK293细胞模型,应用此细胞模型进行辣椒素和甲醛致VR1 -HEK293细胞死亡实验.结果表明,VR1 -HEK293细胞模型构建成功,VR1 细胞死亡实验是一种简便有效的鉴定VR1体外表达的实验方法;甲醛能够激活VR1,且激活反应能被VR1特异性拮抗剂capsazepine所阻断,从而推断甲醛对气道的刺激作用也是通过VR1所介导的.用体外细胞学实验证实了VR1是甲醛作用的生物靶分子,以实验证实了Nielsen假说的正确性.

Construction of VR1+cell model and verification of Nielsen's hypothesis

To explore the physiological role of vanilloid receptor type 1 (VR1) as a possible bio-target molecule of indoor-air pollutant of formaldehyde, VR1 -HEK293 cell model was constructed using the VR1-cDNA plasmid through the molecular cloning and transfection technology. The experiments of capsaicin and formaldehyde inducing death of transfected VR1 -HEK293 cells were carried out. The results showed that VR1 -HEK293 cell model was constructed successfully, and measurement of VR1 -HEK293 cell death rate showed to be an available and effective method, which could be applied to detecting the VR1 expression in cells in vitro. And activated VR1 by formaldehyde could be blocked by capsazepine, a specific antagonist of VR1, Therefore it was suggested that the irritant effect of airway might be also mediated by VR1. Therefore, it was concluded that VR1 as a possible bio-target molecule of formaldehyde, and Nielsen's hypothesis was correct according to our experiments in vitro.