Figure 6.

nNOS mediates the protective effects of aucubin in vitro. (a) and (b) Western blot analysis of nNOS, iNOS, and eNOS in the indicated heart tissue post-MI surgery ((a) representative blots and (b) statistical analysis result) (n = 6, ^∗p < 0.05 versus vehicle-sham and ^#p < 0.05 versus vehicle-MI). (c) and (d) Western blot analysis of nNOS, iNOS, and eNOS in the aucubin- (50 μM) pretreated H9c2 cardiomyocytes after exposure to hypoxia for 24 h ((c) representative blots and (d) statistical analysis result) (n = 4 sample). ^∗p < 0.05 versus normoxia + PBS and ^#p < 0.05 versus hypoxia + PBS. (e) and (f) Cardiomyocytes were pretreated with NAC or aucubin and then stimulated with H₂O₂. (f) Cell viability detected by CKK8 assay in the indicated group (n = 6 samples). (f) ROS level was detected by DCF-DA in the indicated group (n = 6 samples). (g) and (h) Cardiomyocytes were pretreated with L-NAME, L-VINO, or L-canavanine or/and aucubin and then exposed to hypoxia. (g) Cell viability detected by CKK8 assay in the indicated group (n = 6 samples). (h). ROS level was detected by DCF-DA in the indicated group (n = 6 samples). ^∗p < 0.05 versus normoxia + PBS and ^#p < 0.05 versus hypoxia + PBS. All experiments were repeated independently three times.