Effect of exsomes on SH-SY5Y cells after hypoxic ischemia/reperfusion injury

Abstract

Objective To study the effect of exsomes in SH-SY5Y cells after hypoxic ischemia/reperfusion injury. Methods Human umbilical vein endothelial cell hypoxic ischemia/reperfusion (HUVEC I/R) injury models were established, and the exosomes derived from HUVEC I/R were extracted and identified. SH-SY5Y cell hypoxic ischemia/reperfusion injury models (SH-SY5Y I/R) were established, and cells from SH-SY5Y I/R were divided into control group and exosomes-treated group. The proliferation of SH-SY5Y cells was evaluated by CCK-8 assay 24, 48 and 72 h after cell inoculation. Transwell assay and wound-healing assay were used to examine the invasion and migration. Hochest33258 staining and Flow cytometry were used to monitor the changes of cell cycle and apoptosis. Expressions of Caspase-3, Bax and Bcl-2 were measured by real-time fluorescence quantificative-PCR and Western blotting. Results As compared with those in the control group, the proliferation abilities of SH-SY5Y cells in exosomes-treated group were significantly promoted (48 h: 0.70±0.05 vs. 0.94±0.08; 72 h: 0.83±0.05 vs. 1.02±0.06), the cell cycle rate of S phase was significantly increased (14.39%±4.11% vs. 20.54%±3.46%), and G0/G1 phase was statistically decreased (71.26%±5.24% vs. 66.87%±4.23%, P<0.05). What’s more, cell invasive was significantly promoted (44.00±6.56vs. 70.67±6.11), and relative wound injury area was significantly reduced in the exosomes treated group (0.61±0.07 vs. 0.52±0.10); significant differences were noted between the two groups (P<0.05). The mRNA expressions of Bax and Caspase-3 were significantly decreased and the mRNA expressions of Bcl-2 was significantly increased in the exosomes-treated group as compared with those in the control group (P<0.05). Conclusion HUVEC I/R-derived exosomes play neuro-protective role in human SH-SY5Y cells after hypoxic I/R injury. Key words: Exosome; Stroke; Ischemia/reperfusion injury; Neuroprotection