Effects of docosahexaenoic acid on expression of Ang-2 and VEGF in rat brain microvascular endo-thelial cells subjected to oxygen-glucose deprivation / 中华麻醉学杂志

ABSTRACT

Objective To evaluate the effects of docosahexaenoic acid ( DHA) on the expression of angiotensin?2 ( Ang?2) and vascular endothelial growth factor ( VEGF) in rat brain microvascular endo?thelial cells (BMVECs) subjected to oxygen?glucose deprivation (OGD). Methods Primarily cultured rat BMVECs were divided into 3 groups ( n=18 each) using a random number table control group ( group C) , OGD group and DHA group. The cells were cultured with glucose?free and serum?free DMEM culture medium in OGD and DHA groups. In group DHA, DHA 40μmol was added, and then the cells were ex?posed to 1%O2?5%CO2?94%N2 in an air?tight incubator. The cells were cultured in the normal glucose and normal oxygen conditions in group C. All the cells were cultured for 24 h. Cell apoptosis was detected using Annexin V∕propidium iodide flow cytometry assay, and the apoptosis rate was calculated. The concentra?tions of Ang?2, VEGF, prostaglandin E2 ( PGE2 ) and prostacyclin ( PGI2 ) in the supernatant of the cul?ture medium were determined by enzyme?linked immunosorbent assay. The expression of Ang?2 mRNA and VEGF mRNA in cells was detected by real?time polymerase chain reaction. The expression of cyclooxygen?ase?2 ( COX?2) protein in cells was detected by Western blot. Results Compared with group C, the ap? optosis rate and concentrations of Ang?2, VEGF, PGE2 and PGI2 in the supernatant were significantly in?creased, and the expression of Ang?2 mRNA, VEGF mRNA and COX?2 protein was significantly up?regu?lated in OGD and DHA groups (P<0.05). Compared with group OGD, the apoptosis rate and concentra?tions of Ang?2, VEGF, PGE2 and PGI2 in the supernatant were significantly decreased, and the expression of Ang?2 mRNA, VEGF mRNA and COX?2 protein was significantly down?regulated in group DHA ( P<0.05) . Conclusion DHA can inhibit the expression of Ang?2 and VEGF in rat brain BMVECs subjected to OGD and reduce cell apoptosis, and down?regulated expression of COX?2 protein is involved in the mecha?nism.