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Published on: February 28, 2017
[Effect of notch signaling pathway on VEGF promoting rat mesenchymal stem cell proliferation]
Feng-Ling Liao1, Ri-Ling Chen1, Shan Jiang1
1Children's Medical Center of Affiliated Hospital, Guangdong Medical College, Zhanjiang 524000, Guangdong Province, China.
Abstract:
This study was purposed to investigate the effect of Notch signaling pathway on VEGF promoting the proliferation of rat mesenchymal stem cells (MSC). Rat MSC were cultured in vitro, and the cells in logarithmic growth phase were used for experiments. The inhibitor DAPT was used to block Notch signaling pathway, and the effect of the pathway on VEGF promoting proliferation of MSC was observed. The experiment was divided into 4 groups: control, VEGF, DAPT and VEGF+DAPT. The CCK-8 was used to assay the cells proliferation of each group, while RT-PCR was used to detect the changes of related genes (Notch1, Notch2, Flk-1, Hes-1) at mRNA levels. The results indicated that the cells survival rate MSC in DAPT group and VEGF+DAPT group was low in each time point (24 h, 48 h, 72 h), the cell number decreased, and the cells became rounded. The survival rate of MSC in VEGF group was the highest; the difference of cell survival rate was statistically significant between the groups (P < 0.01); Compared with the control group, the mRNA expression level of Notch1, Notch2 and Flk-1 in VEGF group was raised, while the expression level of Notch1 and Notch2 in DAPT group and VEGF+DAPT group come down, with statistically significant differences (P < 0.05); whereas the mRNA expression level of Hes-1 in VEGF group was down-regulated, but that in DAPT group and VEGF+DAPT group was up-regulated, and the difference was statistically significant (P < 0.05). Flk-1 mRNA level in DAPT group and VEGF+DAPT group was slightly lower, but the difference was not statistically significant (P > 0.05). It is concluded that Notch signaling pathway plays an important role in promoting the proliferation of rat MSC, treated with VEGF, however, the DAPT can weaken this effect.
Insights
The Notch signaling pathway is crucial for vascular endothelial growth factor (VEGF)-induced proliferation of rat mesenchymal stem cells (MSCs). Blocking this pathway with DAPT significantly reduces MSC proliferation, indicating its essential role.
Area of Science:
- Cell Biology
- Stem Cell Research
- Molecular Signaling
Background:
- Vascular Endothelial Growth Factor (VEGF) is known to promote cell proliferation.
- The Notch signaling pathway regulates various cellular processes, including stem cell fate and proliferation.
- Understanding the interplay between VEGF and Notch signaling in mesenchymal stem cells (MSCs) is crucial for regenerative medicine.
Purpose of the Study:
- To investigate the role of the Notch signaling pathway in mediating VEGF-induced proliferation of rat mesenchymal stem cells (MSCs).
- To determine how inhibiting the Notch pathway affects VEGF's ability to promote MSC proliferation.
Main Methods:
- Rat MSCs were cultured and treated with VEGF and/or the Notch inhibitor DAPT.
- Cell proliferation was assessed using the CCK-8 assay.
- Gene expression of Notch1, Notch2, Flk-1, and Hes-1 was analyzed at the mRNA level using RT-PCR.
Main Results:
- VEGF treatment significantly increased MSC survival and proliferation compared to the control group.
- Inhibition of the Notch pathway with DAPT reduced MSC survival and proliferation, even in the presence of VEGF.
- VEGF upregulated Notch1, Notch2, and Flk-1 mRNA levels, while downregulating Hes-1.
- DAPT treatment downregulated Notch1 and Notch2, upregulated Hes-1, and had a minor, non-significant effect on Flk-1 mRNA levels.
Conclusions:
- The Notch signaling pathway plays a significant role in promoting VEGF-induced proliferation of rat MSCs.
- Inhibiting the Notch pathway with DAPT attenuates the proliferative effects of VEGF on MSCs.
- These findings highlight the importance of the Notch pathway in MSC proliferation and suggest potential therapeutic targets.
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