SymbolmiR-378 effects on the proliferation and apoptosis of bone marrow mesenchymal stem cells under hypoxic-ischemic condition

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BACKGROUND:To investigate the influence of miR-378 on bone marrow mesenchymal stem cells proliferation and apoptosis under hypoxic-ischemic condition will provide a new method for further improving the survival of bone marrow mesenchymal stem cells in the infarcted myocardium.OBJECTIVE:To observe the viability of bone marrow mesenchymal stem cells and their ability of angiogenesis under hypoxic-ischemic condition after miR-378 transfection.METHODS:The bone marrow mesenchymal stem cells of Sprague-Dawley rat were cultured in vitro and divided into the non-transfection group and the transfection group. Mesenchymal stem cells were transfected with miR-378 mimic in the transfection group while the non-transfection group was taken as the control group. After transfection, the two groups were incubated under hypoxic-ischemic condition (serum-free medium,1% O 2, 5% CO2, 94% N2) at 37 °C for 24 hours. Viability and proliferation of bone marrow mesenchymal stem cells were evaluated by the trypan-blue exclusion assay and MTS assay respectively. The cell apoptosis was detected by TUNEL assay. The culture supernatant of the two groups was collected separately and used as the conditioned medium after their exposure to hypoxic-ischemic environment. Endothelial cells were then co-cultured with the conditioned medium for the vasculature formation assay.RESULTS AND CONCLUSION:After the experience of hypoxia-ischemia for 24 hours and 72 hours, there was a larger number of the living cells in the transfection group in contrast to the non-transfection group (both P < 0.01). Compared with the non-transfection group, miR-378-MSCs group presented a stronger ability of proliferation, and their proliferation rates were obviously higher at 24 and 48 hours (P < 0.01 and P < 0.05, respectively). The percentage of apoptotic cells in the transfection group was significantly declined under the hypoxic-ischemic condition (P < 0.05). The vasculature formation assay indicated that the lumen-like structures were found in both of the two groups, however, the number of lumen-like structures was remarkably increased in the transfection group (P < 0.01). miR-378 could promote the proliferation of bone marrow mesenchymal stem cells and suppress their apoptosis under hypoxic-ischemic condition. It could also enhance their ability of vasculogenesis.Funding:the National Natural Science Foundation of China, No. 81070125, 81270213; the Scientific and Technological Plan of Guangdong Province, No. 2010B031600032; the Fundamental Research Funds for Universities-Key Cultivation Project of Young Teachers in Sun Yat-sen University, No. 13ykzd16

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