无血清条件对诱导转染pEGFP-C1/Akt的骨髓间充质干细胞凋亡的影响

Objective To explore the effect of serum deprivation (SD)-induced apoptosis on mesenchymal stem cells transfected with pEGFP-C1/Akt. Methods The study included experiment group (MSCs transfected with pEGFP-C1/Akt), inhibit group (MSCs transfected by pEGFP-C1/Akt were inhibited by LY294002 20μmol/L) and control group (MSCs). At 4h, 12h, 24h and 36h after SD culture, the expressions of Akt and Caspase-3 mRNA were determined by RT-PCR. Western blot was used for detecting the expression of Akt protein. Immunohistochemistry was used to analyze the expression of Caspase-3 protein. Apoptosis was determined by flow cytometry. Results At 4h, 12h, 24h and 36h after SD culture, the expressions of Akt mRNA and protein had significant elevation in experiment group compared with inhibit group and control group, respectively (P＜0.01). The expressions of Caspase-3 mRNA and protein, and the percentage of apoptosis cells were markedly reduced in experiment group compared with inhibit group and control group, respectively (P＜0.01). Moreover, there were no significant difference between inhibit group and control group shout the expressions of Akt, Caspase-3 and the percentage of apoptosis cells. Conclusion Akt can protect MSCs from SD-induced apoptosis via PI3K Akt and Caspase-3 pathway.