人骨髓间充质干细胞表面CGRP受体存在的研究

目的寻找CGRP受体在MSCs表面存在与否的证据。方法采用梯度离心法和选择性培养基分离、纯化骨髓间充质干细胞,根据分组,培养体系中添加CGRP、CGRP（8-37）。于传代第三代,MTT法检测细胞增殖能力,RT-PCR检测细胞mRNA表达,Western blot检测细胞蛋白表达,以证实MSCs表面是否存在CGRP受体。结果CGRP受体特异性拮抗剂能阻断CGRP对MSCs增殖的促进作用,RT-PCR法证实MSCs表达CGRP受体mRNA,Western blot法证实MSCs表达CGRP受体蛋白。结论MSCs表达CGRP受体,从受体拮抗有效、mRNA表达、蛋白表达角度能得以证实。

Research for Evidence of CGRP Receptor Existence in Human Mesenchymal Stem Cells

Objective To find the evidence of whether the Calcitonin gene-related peptide （CGRP） receptor exists in the mesenchymal stem cells （MSCs）. Methods The human MSCs were isolated from healthy volunteers＇ bone mar rows with gradient centrifugatioru The cells were treated with CGRP or CGRP（8-37） respectively for different groups. Third generation cells were used to detect proliferation by MTT exam, to detect rnRNA expression of CGRP receptor by RTPCR, and to detect protein of CGRP receptor by Western blot. Results Specific antagonists of CGRP Receptor can interdict the of CGRP on MSCs proliferatioru PT-PCR test demonstrated the rnRNA expression of CGRP receptor in MSCs. The Western blot test demonstrated the protein expression of CGRP receptor. Conclusion According to the antagonist test, mRNA test and protein test, it is suggested that CGRP receptor exists in the MSCs.