目的:研究CCK B型受体(cholecystokinin B receptor, CCKBR)与μ型阿片受体(mu opioid receptor, MOR)的异源聚合体对CCKBR功能的调控作用.方法:建立稳定单独转染CCKBR、共同转染CCKBR和MOR的细胞系,通过细胞免疫荧光及Western blotting的方法验证细胞系建立成功,利用不同浓度的CCK-8刺激细胞,检测两种细胞系中受体活化后胞浆钙离子浓度的变化、ERK激酶的活化及细胞增殖活性的检测.结果:相对于单转CCKBR的细胞,稳定双转CCKBR和MOR的细胞中,CCKBR受 CCK-8刺激后引起的胞浆钙离子释放增加,ERK活化增强,细胞增殖速度加快.结论:CCKBR与MOR的异源聚合可增强CCKBR的功能.%Objective: To investigate the influence of heteromerization of cholecystokinin B receptor (CCKBR) and mu opioid receptor (MOR) on the CCKBR' s functions. Methods: HEK293T cells stably transfected with only CCKBR, or both CCKBR and MOR plasmids were used. Expression and membrane localization of the two receptors were examined by immunocytochemical staining and Western blotting, intracellular calcium concentration and ERK activation were used to detect the CCKBR' s function and cell proliferation after CCK-8 stimulation were detected too. Results: In the HEK293T cells, CCKBR and MOR expressed stably and mainly localized on the cell membrane. Compared with the CCKBR single transfected cells, the CCK-8-induced elevation of intracellular calcium concentration and ERK activation in cells co-transfected with both receptor plasmids were significantly stronger. In addition, under the stimulation of CCK-8, the co-transfected cells also proliferated faster. Conclusion: The heteromerization of CCKBR and MOR can enhance the function of CCKBR which may participate in the physiological regulation of opioid and anti-opioid system.
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