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1.
Figure 5

Figure 5. From: Hydrogen Peroxide Inhibits Cytochrome P450 Epoxygenases.

Direct effect of ROS on 3H-14,15-EET in vitro. A through D, Representative HPLC chromatograms following incubation of 3H-14,15-EET with H2O2 or the superoxide-generating system xanthine plus xanthine oxidase (X+XO). 14,15-EET is not directly oxidized by H2O2 or superoxide (n=3). CPM indicates counts per minute. n indicates number of experiments.

Brandon T. Larsen, et al. Circ Res. ;102(1):59-67.
2.
Figure 6

Figure 6. From: Hydrogen Peroxide Inhibits Cytochrome P450 Epoxygenases.

Effect of H2O2 on AA- or 11,12-EET–induced dilation of HCAs. A, AA-induced dilation of endothelium-intact HCAs is reduced by transient exposure to 10−5 mol/L H2O2 (n=7, *P<0.05 vs vehicle), suggesting that H2O2 interferes with the synthesis and/or action of EETs. B, 11,12-EET–induced dilation of endothelium-denuded HCAs is not inhibited by H2O2 (n=6), suggesting that H2O2 does not interfere with the downstream action of EETs on VSMCs. n indicates number of vessels.

Brandon T. Larsen, et al. Circ Res. ;102(1):59-67.
3.
Figure 1

Figure 1. From: Hydrogen Peroxide Inhibits Cytochrome P450 Epoxygenases.

BK-induced dilation of HCAs in the presence of CYP/EET inhibitors or catalase. A, BK induces dilation of HCAs that is not inhibited by sulfaphenazole, EEZE, iberiotoxin, or 6-(2-propargyloxyphenyl)hexanoic acid (PPOH) (n=31, 6, 5, 6, and 7, respectively), indicating little contribution of EETs to this response. B, BK-induced dilation is reduced by catalase (n=31 and 21, respectively; *P<0.05 vs vehicle), indicating that H2O2 partially mediates this response. n indicates number of patients.

Brandon T. Larsen, et al. Circ Res. ;102(1):59-67.
4.
Figure 4

Figure 4. From: Hydrogen Peroxide Inhibits Cytochrome P450 Epoxygenases.

Effect of H2O2 on 14C-AA metabolism by microsomes containing recombinant human CYP2C9 or CYP2J2. A and C, Representative chromatograms following separation of metabolites by reverse-phase HPLC. Migration times of authentic standards are noted on each chromatogram. CPM indicates counts per minute. B and D, EET formation by CYP2C9 and CYP2J2 is inhibited in a concentration-dependent fashion by H2O2 (n=3 to 8 at each concentration; *P<0.05 vs respective vehicle). n indicates number of experiments.

Brandon T. Larsen, et al. Circ Res. ;102(1):59-67.
5.
Figure 2

Figure 2. From: Hydrogen Peroxide Inhibits Cytochrome P450 Epoxygenases.

BK-induced ROS production in isolated HCAs. A, Representative histofluorescence images of HCAs incubated with DHE and DCFH to visualize superoxide and H2O2 production, respectively, in response to BK. Some vessels were denuded of endothelium or treated with catalase. Bar=100 μm. B, BK induces superoxide and H2O2 production in HCAs, as evidenced by the increased ratio of DHE and DCFH fluorescence intensity (I) vs baseline (I0), respectively (n=7). BK-induced ROS production is endothelium dependent (n=4). Catalase inhibits the increase in BK-induced DCFH fluorescence, indicating specificity for H2O2 (n=4). n indicates number of patients. *P<0.05 vs vehicle, †P<0.05 vs BK+vehicle.

Brandon T. Larsen, et al. Circ Res. ;102(1):59-67.
6.
Figure 3

Figure 3. From: Hydrogen Peroxide Inhibits Cytochrome P450 Epoxygenases.

Detection of an EET-mediated component of BK-induced dilation of HCAs in the presence of catalase. A, BK-induced dilation of HCAs in the presence of catalase is inhibited by sulfaphenazole, EEZE, and iberiotoxin (n=21, 5, 5, and 4, respectively; *P<0.05 vs catalase+vehicle), suggesting that EETs mediate BK-induced dilation when H2O2 is reduced. B, Diagram of the dual-cannulated, tandem-perfused HCA bioassay preparation. Endothelium-denuded detector vessels were perfused downstream of endothelium-intact donor vessels. Dilation of detectors was monitored by videomicroscopy. C, Bioassay detection of transferable vasodilators released from HCA endothelium. Donor-applied BK induces dilation of detectors (n=5) in a manner that is partially inhibited by donor-applied catalase (n=5) but not by detector-applied EEZE (n=5). However, an inhibitory effect of detector-applied EEZE is unmasked in the presence of donor-applied catalase (n=5), suggesting that EETs represent transferable vasodilators, the release of which is enhanced when H2O2 is reduced. n indicates number of patients. *P<0.05 vs BK+vehicles, †P<0.05 vs BK+donor catalase+detector vehicle.

Brandon T. Larsen, et al. Circ Res. ;102(1):59-67.

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