[Regulation of sonic hedgehog on vascular endothelial growth factor, basic fibroblast growth factor expression and secretion in bone marrow mesenchymal stem cells]

Jiaqin Cai, Yizhou Huang, Xiaohe Chen, Honglei Xie, Yongcan Huang, Li Deng
Chinese Journal of Reparative and Reconstructive Surgery 2012, 26 (1): 112-6

OBJECTIVE: Sonic hedgehog (Shh) signaling pathway is involved in an important part of regulating angiogenesis. To investigate the effects of recombinant Shh N-terminate (rShh-N) on the expression and secretion of angiogenesis-related factor-vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF).

METHODS: Bone marrow mesenchymal stem cells (BMSCs) were isolated from 3-day-old healthy Sprague Dawley rats and cultured to passage 3 in vitro. rShh-N at the concentrations of 0, 10, 100, and 200 ng/mL were applied to culture BMSCs in groups A, B, C, and D, respectively. At 12, 24, 48, and 72 hours of culture, the expressions of VEGF and bFGF mRNA and the levels of VEGF and bFGF in supernatant were measured with real-time quantitative PCR and ELISA, respectively.

RESULTS: At the gene level, compared with group A, the expressions of VEGF and bFGF mRNA were enhanced in group D (P < 0.05) and the upregulation was more significant at 12 and 48 hours than 24 and 72 hours (P < 0.01). In group C, bFGF mRNA expression was substantially promoted at 12-72 hours (P < 0.05) and VEGF mRNA level was upregulated at 24-72 hours (P < 0.05), and both reached peak at 72 hours (P < 0.01). In group B, VEGF mRNA expression was inhibited at 12 hours (P < 0.05), but the level increased at 48 and 72 hours (P < 0.05); bFGF mRNA expression was obviously promoted at 12-48 hours (P < 0.05) and the maximum appeared at 48 hours (P < 0.01). At the protein level, the secretion of VEGF and bFGF in group D was significantly increased at 12-72 hours, as compared with group A (P < 0.05). In group C, VEGF and bFGF secretion was increased at 24-72 hours (P < 0.05). The secretion of VEGF in group B was inhibited at 12 and 48 hours (P < 0.05) and was promoted at 24 hours (P < 0.05); bFGF secretion was up-regulated at 24 and 48 hours (P < 0.05). The secretion of VEGF and bFGF in supernatant at 48 and 72 hours were significantly more than those at 12 and 24 hours in 4 groups (P < 0.05).

CONCLUSION: rShh-N treatment can enhance the expression and secretion of VEGF and bFGF in BMSCs, which could provide the experimental evidence for the further application of Shh-MSCs in the treatment of ischemia-related diseases and bone repair.

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