Xingnaojing Injection Protects against Cerebral Ischemia Reperfusion Injury via PI3K/Akt-Mediated eNOS Phosphorylation

Figure 6

XNJ induced endothelial NO synthase (eNOS) phosphorylations and nitric oxide (NO) enhancement in HBMECs after exposure to “OGD”. HBMECs were treated with XNJ (1.5μl/ml, 2.5μl/ml) and received 3 h OGD followed by 24 h recovery. LY294002 (10μM) was added to XNJ (2.5μl/ml) group. (a) The protein expression of phosphorylated eNOS (p-eNOS) at Ser-1177 and eNOS was analyzed by western blot; (b) p-eNOS was upregulated in the HBMECs exposed to OGD pretreated with XNJ (2.5μl/ml) by immunofluorescent staining. Positive stainings as shown in green. Scale bars: 50 μm. (c)NO production in supernatant was assayed by Griess method. LY294002 (10μM) pretreatment alleviated the XNJ induced eNOS phosphorylation and NO production. Data were expressed as means ± SD (n = 4). p < 0.001 vs. CTL; #p < 0.05 vs. OGD group; ##p < 0.01 vs. OGD group; ###p < 0.001 vs. OGD group; & p < 0.05 vs. OGD +XNJ (2.5μl/ml) group; && p < 0.01 vs. OGD +XNJ (2.5μl/ml) group.

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