The effect of calcitonin gene-related peptide on proliferation and migration of human umibilical vein and its mechanisms
Objective To investigate the effect of calcitonin gene-related peptide(CGRP)on proliferation and migration of human umbilical vein endothelial cells(HUVECs)and the underlying mechanisms.Methods HUVECs were cultured and randomly divided into control group,CGRP group,bevacizumab(VEGF inhibitor)group,and L-NMMA(nitric oxide synthase inhibitor)group.The MTT assay was employed to evaluate the effects of CGRP,bevacizumab and L-NMMA on the viability of HUVECs.The scratch assay was conducted to assess the migration of HUVECs and the expression levels of relevant proteins were detected by immunocytochemistry assay and Western blot.Results CGRP treatment markedly promoted the proliferation of HUVECs by binding to its receptor,calcitonin recep-tor-like receptor(CRLR)(t=3.434,P＜0.05).When VEGF or eNOS was inhibited,the CGRP-induced endothelial cell proliferation was significantly suppressed,as indicated by decreased cell viabilities(P＜0.05)and the expression of cell makers(P＜0.05).In the CGRP stimulated HUVECs,inhibition of VEGF or eNOS could also suppress cell migration as detected by the scratch assay(P＜0.05).Additionally,inhibition of VEGF or eNOS decreased the expression of the Raf-1,a kinase that is closely related to proliferation(P＜0.05).It also reduced the activities of its downstream extracellular-signal-regulated kinase(ERK)(P＜0.05)and c-Jun N-terminal kinase(JNK)(P＜0.05)signaling pathways in HUVECs stimulated by CGRP.Conclusion CGRP may induce HUVECs proliferation and migration through activating the VEGF/eNOS-Raf-1-ERK/JNK signaling pathway,suggesting that inhibition of VEGF or eNOS may serve as a potential target for anti-angiogenic therapy.
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维普
