[Calcitonin gene-related peptide promoting migration of rat bone marrow mesenchymal stem cells and stimulating expression of vascular endothelial growth factor].
OBJECTIVE
To explore the effects of calcitonin gene-related peptide (CGRP) on the migration of bone marrow mesenchymal stem cells (BMSCs) and vascular endothelial growth factor (VEGF) expression in vitro.


METHODS
The BMSCs were isolated from Sprague Dawley rats using whole bone marrow adherence method. At 1, 2, and 3 weeks after culture, the expressions of CGRP receptor (CGRPR) was detected by Western blot. The BMSCs were treated with CGRP at concentration 1 x 10(-8) mol/L (experimental group) and did not treated (control group), and the efficacy of BMSCs migration was analyzed by Transwell chamber assay after 72 hours; at 1, 3, 5, and 7 days, the mRNA expressions of vascular cell adhesion molecule 1 (VCAM-1) were detected by real-time fluorescent quantitative PCR; the protein expressions of VEGF were examined using immunohistochemistry and Western blot.


RESULTS
CGRPR expressed stably in the cultured BMSCs and reached the peak at 2 weeks. CGRP had a significantly enhanced role in promoting cell migration. The number of cell migration was (3.20 +/- 1.77) cells/HP in experimental group and (1.11 +/- 0.49) cells/HP in control group, showing significant difference (t = 4.230, P = 0.001). In experimental group, the expressions of VCAM-1 mRNA increased with time and reached the peak at 7 days. There were significant differences in the expressions of VCAM-1 mRNA between control group and experimental group at 3, 5, and 7 days (P < 0.05). Immunocytochemistry results showed positive DAB staining for VEGF at 5 and 7 days in experimental group. Western blot results showed that the protein expressions of VEGF increased significantly at 5 and 7 days in experimental group when compared with control group (P < 0.05), which was signfiantly higher at 5 days than at 7 days in experimental group (P < 0.05).


CONCLUSIONS
CGRP can promote the migration of BMSCs and stimulate the protein expression of VEGF, which may plays an important role in regulating bone metabolism by increasing angiogenesis.
Department of Orthopaedics and Traumatology, Nanfang Hospital, Southern Medical University, Guangzhou Guangdong 510515, PR China.
Zhao
Z
Wang
Dan
D
Jin
Yonghua
Y
Tuo
Xiaolei
X
Guo
Jun
J
Wen
Jian
J
Zhou
Liheng
L
Xia
Yongtao
Y
Zhang
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology / Q000378:metabolism*; D015740:Calcitonin Gene-Related Peptide; D002465:Cell Movement; D002478:Cells, Cultured; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D018015:Receptors, Calcitonin Gene-Related Peptide; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*
D018015:Receptors, Calcitonin Gene-Related Peptide; D042461:Vascular Endothelial Growth Factor A; C467498:vascular endothelial growth factor A, rat; D015740:Calcitonin Gene-Related Peptide