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Experimental Biology and Medicine 228:454-458 (2003)
© 2003 Society for Experimental Biology and Medicine

HEME OXYGENASE

Functional Expression of Human Heme Oxygenase-1 Gene in Renal Structure of Spontaneously Hypertensive Rats

Alvin I. Goodman{dagger}, Shou Quan*, Liming Yang*, Arika Synghal{dagger} and Nader G. Abraham*,1

* Department of Pharmacology and
{dagger} Division of Nephrology, Department of Medicine, New York Medical College, Valhalla, New York 10595

Abstract

Heme oxygenase (HO), by catabolizing heme to bile pigments, regulates the levels and activity of cellular hemoprotein and HO activity. We examined the effect of delivery of the human HO-1 gene on cellular heme in renal tissue using a retroviral vector. We used a single intracardiac injection of the concentrated infectious viral particles in 5-day-old spontaneously hypertensive rats; 25 were transduced with empty vector and 25 were transduced with the human HO-1 gene. Functional expression of human and rat HO-1 was measured after 2 and 4 weeks. Reverse transcription polymerase chain reaction showed that human HO-1 mRNA was expressed as early as 2 weeks, with the highest levels in the kidney. Western blot analysis showed distribution of human HO-1 protein in rat kidney structures, predominantly in the thick ascending limb of the loop of Henle as well as in proximal tubules and preglomerular arterioles. These areas also demonstrated higher HO activity as measured by increased conversion of heme to bilirubin and carbon monoxide. Functional expression of the human HO-1 gene was associated with a decrease in blood pressure in 4- and 8-week-old spontaneously hypertensive rats. Compared with nontransduced rats, human HO-1 gene overexpression in transduced rats was associated with a 35% decrease in urinary 20-hydroxyeicosatetraenoic acid, a potent vasoconstrictor and an inhibitor of tubular Na+ transport, which may be related to the decrease in blood pressure.

Key Words: heme oxygenase • cytochrome P450 • hypertension

The kidney plays a central role in the regulation of salt and water balance affecting the control of blood pressure (BP). Fluid and electrolyte homeostasis by this organ involves different mechanisms, including the localized formation of substances that affect renal tubular function and/or blood flow, such as 20-hydroxyeicosatetraenoic acid (20-HETE), an {omega}-hydroxylated metabolite of arachidonic acid (AA), which has been shown to be a potent inhibitor of renal tubular Na+/K+-ATPase activity as well as a powerful constrictor of kidney microvessels (1). Renal excretion of 20-HETE is reciprocally related to the expression of heme oxygenase-1 (HO-1)(2) and reduction in its production or the increase of the latter may favor, at least in part, the lowering of BP in spontaneously hypertensive rats (SHRs). Because 20-HETE promotes vasoconstriction at renal and extrarenal sites, it may be a mediator of prohypertensive mechanisms in SHRs, in which 20-HETE production was reported to be increased (3). The {omega}-hydroxylation of AA to 20-HETE is catalyzed by enzymes of the cytochrome P-450 monooxgenases 4A isoform (CYP4A) gene family. This subfamily encodes several CYP enzymes that are capable of hydroxylating the terminal {omega}-carbon and, to a lesser extent, the {omega}-1 hyproxylation of saturated and unsaturated fatty acids as well as various prostaglandins (4). In CYP enzymes, the heme sits deep inside the protein with its iron ligated to a cysteine thiolate group and this coordination of arrangement gives CYP enzymes their unique spectral and catalytic properties.

Heme and products derived from its metabolism potentially influence renal function and BP by affecting the expression of and/or activity of hemoproteins, including CYP and cyclooxygenases (COX-1 and COX-2). One of the metabolites of heme, carbon monoxide (CO), has been shown to function as a vasodilator. It has also been documented that increased expression of HO attenuates reactivity to constrictor agonists and that HO inhibitors magnify myogenic tone in gracilis muscle arterioles (5). Recent studies have also indicated that administration of HO inhibitors increases arterial pressure in normotensive rats (6). These observations suggest that endogenous HO-derived CO probably plays a role in the regulation of basal arterial tone and, thus, contributes to modulating BP (7). Induction of HO-1 by heavy metals, or by its substrate heme (8, 9), was shown to increase HO activity and decrease BP in SHRs, thus, connotating a reciprocal relationship between HO gene expression and hypertension in SHRs (9–12). A previous study showed high levels of HO-1 in isolated proximal tubules and high HO-2 levels in the thick ascending limb of the loop of Henle (mTALH) and in preglomerular arterioles (10). In vivo administration of HO-1 and HO-2 antisense oligodeoxynucleotides further confirmed that HO-2, but not HO-1, contributed to the basal HO activity (9).

This study was designed, first, to examine nonchemical means to increase HO-1 and the suitability of human HO-1 gene transfer delivery into rat tissue and observe its functional expression and, to characterize its effect on cellular heme in renal structures. Our data demonstrate that selective delivery of the human HO-1 gene resulted in an increase of HO activity and a significant decrease in cellular heme. The presence of the human HO-1 gene in the renal structure and vessels of SHRs was associated with an increase in urinary excretion of 20-HETE.

Materials and Methods

Preparation of Concentrated Viral Particles.
The human heme oxygenase-1 (HHO-1)-expressing replication-deficient retrovirus vector LSN-HHO-1 was constructed as previously described (12). Viral particles from supernatants of retroviral packing cells (PT67/HOP-HHO-1) were sedimented by centrifugation at 9000g for 12–14 hr at 4°C in a 45 ml sterile centrifuge tube. After centrifugation, the supernatant was removed by aspiration and the pellet was resuspended in 1 ml cell culture media or Hank’s balanced salt solution. Retroviral particles, sedimented as described above, were further concentrated by centrifugation at 10,000g, 4°C for 12 hr in a 1.5-ml tube. Then, the supernatant was removed and the pellet was resuspended in 20–50 µl of physiological saline solution (13).

Animal Treatment.
Pregnant SHR were purchased from Taconic Laboratories (Germantown, NY). Five-day-old SHRs were divided into two treatment groups: control and LSN-human HHO-1. Treatments were administered by bolus injection of 10 µl of Hanks Balanced Salt Solution with or without LSN-human HHO-1 viral particles (1 x 1010 CFU/ml) directly into the left ventricle under methoxyflurane anesthesia as previously described (12). To determine whether the human HO-1 promoter can be effective in driving the HO-1 gene in vivo, we injected newborn SHRs with concentrated retroviruses via the cardiac route. Newborn rats were chosen because their cells have a stronger ability to proliferate and differentiate than those of adult rats (14) and retroviruses only infect actively dividing cells. The pups were injected with 20 and 40 µl of retrovirus at Day 5 and Day 12, respectively. After injection, the rats were returned to cages with their mothers for continued weaning until weeks 2 and 4. There was a 94.5% survival rate 48 hr after LSN-human HO-1 administration. At different time points, rats were taken for measurement of human HO-1 expression by reverse transcription polymerase chain reaction (RT-PCR). Animals were weaned at 21 days of age; males were separated and used for all experiments. Systolic blood pressure was measured by tail cuff sphygmomanometry twice weekly. Starting at 4 and 8 weeks of age, rats were sacrificed at different times and tissues, including kidney, liver, lung, brain, and aorta, were isolated for determination of human HO-1 expression and HO activity.

Isolation of Proximal Tubule, mTALH, and Medullary Collecting Duct Segments (IMCD).
Proximal tubules were isolated by Percoll gradient separation. This method yields about 6–8 mg of microsomal protein from one rat (two kidneys). mTAL and IMCD segments were isolated from the inner strip of the outer medulla and the inner medulla, respectively, using enzymatic digestion followed by a sieving technique as previously described (2).

RT-PCR and PCR Analysis.
PCR was performed using a Taq PCR kit (Roche, Indianapolis, IN). For each RT-PCR, a sample without reverse transcriptase was processed in parallel and served as a negative control. Cycling parameters for amplifying RT products were as follows: 95°C, 1 min; 60°C, 1 min; 72°C, 1–3 min, for 30 cycles, and then extended at 72°C for another 5 min. After amplification, PCR products were electrophoresed on 1.2% agarose gel, stained with ethidene bromide, and visualized under UV light (11).

Western Blot Analysis.
Tissue homogenates were collected for Western blot analysis. For detection of human and rat HO-1- and HO-2-immunoreactive proteins, 20 µg of tissue homogenate was electrophoresed on a 12% polyacrylamide gel. Specific bands corresponding to HHO-1, rat HO-1 and rat HO-2 proteins were identified with the use of specific monoclonal antibodies (Stressgen Biotechnology, Vancouver, Canada).

Urine Extraction.
Urine was collected for 7 days. 3H-29-HETE (50,000 cpm) and [20,20-2H2] 20-HETE (100 ng) were added to 20 ml of the pooled urine. The sample was adjusted to pH 8.0 with 1 N NaOH and allowed to stand at room temperature for 15 min. After acidification to pH 3.0 with 0.1 N HCl, the sample was applied onto a Tox Elut column (Analytical International, Harbor City, CA) and eluted three times with 20 ml of dichloromethane. The combined organic extract was washed with 0.05 N sodium borate buffer (pH 8.0) and evaporated to dryness. The residue was resuspended in 200 µl of methanol and lipids were separated by thin-layer chromatography on silica gel G using the upper phase of ethyl acetate:water:iso-octane:acetic acid (110:100:50:20, v/v/v/v). Radioactivity was monitored by a radioactive scanner and the zone corresponding to the 20-HETE standard was scraped and extracted with ethyl acetate. The final extract was evaporated and incubated with 100 µl of N-methyl-N-(tert-butyldimethysilsilyl)-trifluoroacetamide for 30 min at 65°C to obtain the PFB ester, tert-butyldimethylsilyl (TBDMS) ether derivative of 20-HETE.

Gas-Chromatography/Mass-Spectometry (GC/MS).
Negative chemical ionization–GC/MS was performed using a mass spectrometer (HP 5989A, Hewlett Packard, Palo Alta, CA) interfaced with a SP-2330 capillary column (15 m x 0.25 mm inside diameter, 0.2-µm film thickness; Supelco, Bellefonte, PA) programmed from 100–250°C at 25°C/min using helium as the carrier gas. An estimation of the total 20-HETE content in the purified biological sample was made by comparing the ion intensities at m/z 433:435 versus a standard curve of 20-HETE-PFB-TBDMS/2H2-20-HETE-PFB-TBDMS molar ratio constructed by negative chemical ionization–GC/MS analysis.

Measurement of HO Activity.
Renal tissue homogenate HO activity was assayed by the method of Abraham et al. (17) in which bilirubin, the product of HO degradation, was extracted with chloroform and its concentration was determined spectrophotometrically by using the difference between absorbance at wavelength 460 nm and then at 530 nm with an absorption coefficient of 40 mM-1•cm-1 (18).

Microsomal Heme Determination.
Microsomal heme was determined as the pyridine hemochromogen by using the reduced minus oxidized difference in absorbance at 400 and 600 nm with an absorption coefficient of 32.4 mM-1•cm-1.

Results

Expression of Human HO-1 in SHR Tissues.
The results showed that 100% (10/10) of the rats injected with retrovirus-mediated human HO-1 driven by the human HO-1 promoter expressed human HO-1 mRNA in all tissues examined. However, in rats injected with empty vector, no RT-PCR band was detected in tissues obtained at 2 weeks (data not shown) or in tissues obtained after 4 weeks (Fig. 1BGo, Lane 4). These data indicate that the human HO-1 gene can be efficiently delivered and expressed in newborn rats via retroviruses, and that the human HO-1 promoter can execute its function to drive the human HO-1 gene expression in vivo.



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  		Figure 1. Detection of human HO-1 transcripts by RT-PCR analysis in 2-week-old (A) and 4-week-old (B) SHRs. Newborn rats were injected twice (Days 5 and 12) with concentrated retrovirus (LSN-human HO-1). Rats were taken at different times and their tissues were prepared for RT-PCR analysis. Human HO-1 transcripts were detected after both 2 weeks and 4 weeks in the kidney, liver, lung, spleen, and heart of rats injected with LSN-HHO-1. RT-PCR shows the quality of RNA was the same in all preparations as evidenced from the equal density of the actin RT-PCR band (C).

 
Effect of Human HO-1 Gene Transduction on Expression of Human HO-1 Protein.
The functional expression of human HO-1 gene transfer in SHRs was evaluated by the levels of human HO-1 protein in renal structures. Western blot analyses of cortical, outer medullary, inner medullary/papillary, and whole kidney microsomal preparations indicated low basal levels of HO-1 protein, which that were markedly increased following treatment with the human HO gene (Fig. 2Go). The increase in HO-1 protein levels was most prominent in the microvessels, followed by the inner medulla/papilla and the aorta (Fig. 2Go). Unlike HO-1, HO-2 protein levels did not significantly change in retrovirus-mediated human HO-1 gene transfer treated rats. HO-2 was detected in all kidney structures. It is interesting to note that the retroviral vector delivered higher levels of human HO-1 to renal microvessels than to other renal segments at both 2 and 4 weeks.



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  		Figure 2. HHO-1 protein distribution in rat kidney. Rat tissue samples were evaluated for immunoreactive rat and human HO-1 protein levels by Western blot analysis. Human HO-1 was at the highest levels in microvessels at both 2 and 4 weeks after retrovirus injection. However, a higher expression was seen at 4 weeks in all tissues tested. K, kidney; IM, inner medulla; OM, outer medulla; MV, microvessels; and C, positive control.

 
Effect of Human HO-1 Gene Transfer on Blood Pressure and 20-HETE Production.
The injection of SHRs with the retrovirus vector mediated human HO-1 gene was associated with a marked decrease in BP compared with SHRs injected with the empty vector. BP of SHRs transduced with human HO-1 gene (n = 6) was 160 ± 7 mmHg compared with187 ± 5 mmHg in SHRs transduced with empty vector (n = 6; P < 0.005) at 4 and 8 weeks after the virus injection (Fig. 3AGo). There was a similar decrease in BP in SHRs receiving LSN-human HO-1 after 4 weeks compared with control SHRs.



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  		Figure 3. (A) Effect of LSN-human HO-1 treatment on BP in SHRs. At 5 days of age, animals were injected with vehicle solution or LSN-human HO-1 (1 x 1010 CFU/ml). BP was determined twice weekly by the tail cuff method. BP of LSN-human HO-1 treated rats (n = 32) was significantly different (*P < 0.001) from vehicle-treated SHR (n = 10) or LSN-treated rats (n = 23) as analyzed by ANOVA and the Student’s t test. (B) Urinary excretion of 20-HETE in SHRs injected with LSN-HHO-1 or vehicle solution. *P < 0.05.

 
Because the development of hypertension in SHRs has been linked to increased expression of a prohypertensive mechanism mediated by elevation of cellular heme, CYP4A and, subsequently, 20-HETE synthesis, we compared urinary excretion of 20-HETE in rats injected with the retrovirus-mediated human HO-1 gene and rats injected with empty vector. As seen in Figure 3BGo, 24-hr urinary excretion of 20-HETE in 7-week-old SHRs treated with LSN-human HO-1 was lower (P < 0.01) than that of SHRs receiving the empty vector (1.29 ± 0.06 and 2.2 ± 0.06 ng 20-HETE per milliliter in rats treated with LSN-human HO-1 and vehicle, respectively, n = 6).

Effect of Differential Expression of Human HO-1 Gene Transfer on HO Activity and Heme in Renal Structures.
The efficacy of LSN-human HO-1 gene transfer in SHRs was also evaluated by comparing the levels of HO activity in different segments of the kidney. As seen in Figure 4Go, the basal levels of HO activity in the outer medulla and the inner medulla/papilla were, respectively, 23% and 56% higher compared with the cortex (P < 0.05). To further ascertain the metabolic characteristics of the different renal structures expressing human HO-1, we assessed the levels of cellular heme. As seen in Figure 5Go, heme content was decreased 15% in the inner medulla/papilla compared to 25% in the cortex (P < 0.05). These findings indicate that retrovirus-mediated human HO-1 gene transduction into SHRs generated an increase in the levels of HO-1 activity in different renal segments. Western blot analysis showed that functional expression of human HO-1 by gene transfer did not modulate HO-2 expression.



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  		Figure 4. Effect of retrovirus-mediated human HO-1 gene transfer on heme content in different renal segments. Heme content was measured as described in Material and Methods. *P < 0.05 versus control tissues. K, kidney; mTALH, thick ascending limb of loob of Henle; IM, inner medulla; and PT, proximal tubule. Values are expressed as means ± SE of three different experiments.

 


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  		Figure 5. Effect of retrovirus-mediated human HO-1 gene transfer on HO activity in different renal segments. HO activity was measured as described in Materials and Methods. *P < 0.05 versus control tissues. K, kidney; mTALH, thick ascending limb of loob of Henle; OM, outer medulla; and PT, proximal tubule. Values are expressed as means ± SE of three different experiments.

 
Discussion

The present study demonstrates, that administration of a retrovirus-mediated human HO-1 gene resulted in a differential expression of human HO-1 protein in renal tissue. The expression of the endogenous rat constitutive HO-2 and the inducible HO-1 proteins were not modulated by delivery of the human HO-1 gene. Our study demonstrates that delivery of the human HO-1 gene enhanced total HO activity at different levels in renal tissues and showed a concomitant decrease in cellular heme and CYP4A-mediated AA metabolism to 20-HETE.

The fact that hypertension is attenuated in SHRs expressing the human HO-1 gene implies that a mechanism dependent on the function of this human gene transfer is related to the lowering effect on BP. Thus, the attenuating influence of LSN-human HO-1 treatment on the development of hypertension in SHRs is most likely the consequence of increased HO activity. This is in agreement with reports that chemical interventions that increase HO activity in SHRs also attenuate the development of hypertension (4).

The mechanism by which human HO-1 gene delivery into rats increases HO activity and decreases BP has not been clearly documented. One explanation may be caused by the increase in CO levels generated in rat overexpression of the HO-1 gene and increase heme degradation. It has been documented that CO, arising from heme metabolism by HO, exerts a vasodilatatory effect (9, 16); that increased ex-pression of HO attenuates reactivity to constrictor agonists (20, 21); and that CO stimulates the apical 70-pS K-channel of the mTALH (22). Another proposed mechanism for the lowering effect of BP as a result of human HO-1 gene expression might be a decrease in cellular CYP4A with subsequent decrease in urinary 20-HETE (15, 20, 21, 23). In support of our hypothesis, it has been shown that induction of HO-1 by heavy metals, such as SnCl2 or heme analogues, results in a depletion of several hemoproteins, including CYP4A, (24, 25). This effect was associated with a selective decrease in renal CYP-AA metabolism and a concomitant reduction in BP, suggesting a prohypertensive role for CYP-AA metabolites, especially in the generation of 20-HETE (26, 27). An additional possible antihypertensive process linked to the induction of HO-1 in SHRs is the enhanced degradation of heme to biliverdin, which, along with bilirubin, is endowed with antioxidant activity. A role of reactive oxygen species (ROS) in the pathogenesis of hypertension have been demonstrated (28) and the elevation of bilirubin may attenuate ROS and lower BP. The delivery of the human HO-1 gene and elevation of bilirubin synthesis in renal structures may decrease blood pressure in SHRs by interfering with oxidative stress (28).

In conclusion, this study demonstrates that the delivery and distribution of the human HO-1 gene into different kidney structures contributes to a decrease in cellular heme and a decrease in BP in SHRs. Furthermore, this study suggests that the delivery of the HO-1 gene in a site-specific manner, may be of significance in the control of renal heme and 20-HETE synthesis.

Acknowledgments

We thank Ms. Sylvia Shenouda for her technical assistance and Ms. Jennifer Brown for her excellent secretarial assistance. We thank Dr. Giovanni LiVolti for computer assistance and rearranging various figures.

Footnotes

This work was supported by the National Institutes of Health Grants HL34300 and DK56601, and American Heart Association Grants 50948T and 20285T supported, in part, by a grant from the Westchester Artificial Kidney Foundation, Inc.

1 To whom requests for reprints should be addressed at Department of Pharmacology, New York Medical College, Valhalla, NY 10595. E-mail: nader_abraham{at}nymc.edu Back

References

  1. Ma YH, Gebremedhin D, Schwartzman ML, Falck JR, Clark JE, Masters BS, Harder DR, Roman RJ. 20-Hydroxyeicosatetraenoic acid is an endogenous vasoconstrictor of canine renal arcuate arteries. Circ Res 72:126–136, 1993.[Abstract/Free Full Text]
  2. Botros FT, Schwartzman ML, Abraham NG. Regulation of cyclooxygenase- and cytochrome P450-derived eicosanoids by heme oxygenase isoforms in rat kidney. Hypertension 38:505–506, 2001.
  3. Omata K, Abraham NG, Escalante B, Schwartzman ML. Age-related changes in renal cytochrome P-450 arachidonic acid metabolism in spontaneously hypertensive rats. Am J Physiol 262:F8–F16, 1992.
  4. Laniado-Schwartzman M, Abraham NG. The renal cytochrome P-450 arachidonic acid system. Pediatr Nephrol 6:490–498, 1992.[Medline]
  5. McCoubrey WK Jr, Huang TJ, Maines MD. Isolation and characterization of a cDNA from the rat brain that encodes hemoprotein heme oxygenase-3. Eur J Biochem 247:725–732, 1997.[Medline]
  6. Kozma F, Johnson RA, Zhang F, Yu C, Tong X, Nasjletti A. Contribution of endogenous carbon monoxide to regulation of diameter in resistance vessels. Am J Physiol 276:R1087–R1094, 1999.
  7. Zhang F, Kaide JI, Rodriguez-Mulero F, Abraham NG, Nasjletti A. Vasoregulatory function of the heme-heme oxygenase-carbon monoxide system. Am J Hypertens 14:62S–67S, 2001.[Medline]
  8. Goodman AI, Choudhury M, da Silva JL, Jiang S, Abraham NG. Quantitative measurement of heme oxygenase-1 in the human renal adenocarcinoma. J Cell Biochem 63:342–348, 1996.[Medline]
  9. Johnson RA, Lavesa M, Askari B, Abraham NG, Nasjletti A. A heme oxygenase product, presumably carbon monoxide, mediates a vasodepressor function in rats. Hypertension 25:166–169, 1995.[Abstract/Free Full Text]
  10. da Silva JL, Zand BA, Yang LM, Sabaawy HE, Lianos E, Abraham NG. Heme oxygenase isoform-specific expression and distribution in the rat kidney. Kidney Int 59:1448–1457, 2001.[Medline]
  11. Ito O, Alonso-Galicia M, Hoop KA, Roman RJ. Localization of cytochrom P4504A isoforms along the rat nephron. Am J Physiol 274: F395–F404, 1998.
  12. Yang L, Quan S, Abraham NG. Retrovirus-mediated HO gene transfer into endothelial cells protects against oxidant-induced injury. Am J Physiol 277:L127–L133, 1999.
  13. Sabaawy HE, Zhang F, Nguyen X, Elhosseiny A, Nasjletti A, Schwartzman M, Dennery P, Kappas A, Abraham NG. Human heme oxygenase-1 gene transfer lowers blood pressure and promotes growth in spontaneously hypertensive rats. Hypertension 38:210–215, 2001.[Abstract/Free Full Text]
  14. Iyer SN, Lu D, Katovich MJ, Raizada MK. Chronic control of high blood pressure in the spontaneously hypertensive rat by delivery of angiotensin type 1 receptor antisense. Proc Natl Acad Sci USA 93:9960–9965, 1996.[Abstract/Free Full Text]
  15. Coceani F, Kelsey L, Seidlitz E, Marks GS, McLaughlin BE, Vreman HJ, Stevenson DK, Rabinovitch M, Ackerley C. Carbon monoxide formation in the ductus arteriosus in the lamb: Implications for the regulation of muscle tone. Br J Pharmacol 120:599–608, 1997.[Medline]
  16. Johnson RA, Colombari E, Colombari DSA, Lavesa M. Role of endogenous carbon monoxide in central regulation of arterial pressure. Hypertension 30:962–967, 1997.[Abstract/Free Full Text]
  17. Abraham NG, Lin JH, Dunn MW, Schwartzman ML. Presence of heme oxygenase and NADPH cytochrome P-450 (c) reductase in human corneal epithelium. Invest Ophthalmol Vis Sci 28:1464–1472, 1987.[Abstract/Free Full Text]
  18. Chernick RJ, Martasek P, Levere RD, Margreiter R, Abraham NG. Sensitivity of human tissue heme oxygenase to a new synthetic metalloporphyrin. Hepatology 10:365–369, 1989.[Medline]
  19. Abraham NG, Mieyal PA, Quan S, Yang L, Burke-Wolin T, Mingone CJ, Goodman AI, Nasjletti A, Wolin MS. Modulation of cyclic GMP by retrovirus-mediated human heme oxygenase-1 gene transfer in microvessel endothelial cells. Am J Physiol 283:L1117–L1124, 2002.
  20. Sammut IA, Foresti R, Clarck JE, Exon DJ, Vesely MJJ, Sarathchandra P, Green CJ, Motterlini R. Carbon monoxide is a major contributor to the regulation of vascular tone in aortas expressing high levels of haeme oxygenase-1. Br J Pharmacol 125:1437–1444, 1998.[Medline]
  21. Kaide JI, Zhang F, Wei Y, Jiang H, Yu C, Wang WH, Balazy M, Abraham NG, Nasjletti A. Carbon monoxide of vascular origin attenuates the sensitivity of renal arterial vessels to vasoconstrictors. J Clin Invest 107:1163–1171, 2001.[Medline]
  22. Liu H, Mount DB, Nasjletti A, Wang W-H. Carbon monoxide stimulates the apical 70-pS K+ channel of the rat thick ascending limb. J Clin Invest 103:963–970, 1999.[Medline]
  23. Christodoulides N, Durante W, Kroll MH, Schafer AI. Vascular smooth muscle cell heme oxygenases generate guanylyl cyclase-stimulatory carbon monoxide. Circulation 91:2306–2309, 1995.[Abstract/Free Full Text]
  24. Abraham NG, Lin JH, Schwartzman ML, Levere RD, Shibahara S. The physiological significance of heme oxygenase. Int J Biochem 20:543–558, 1988.[Medline]
  25. Levere RD, Martasek P, Escalante B, Schwartzman ML, Abraham NG. Effect of heme arginate administration on blood pressure in spontaneously hypertensive rats. J Clin Invest 86:213–219, 1990.
  26. Laniado-Schwartzman M, Lavrovsky Y, Stoltz RA, Conners MS, Falck JR, Chauhan K, Abraham NG. Activation of nuclear factor kappa B and oncogene expression by 12(R)-hydroxyeicosatrienoic acid, an angiogenic factor in microvessel endothelial cells. J Biol Chem 269:24321–24327, 1994.[Abstract/Free Full Text]
  27. Martasek P, Schwartzman ML, Goodman AI, Solangi KB, Levere RD, Abraham NG. Hemin and L-arginine regulation of blood pressure in spontaneous hypertensive rats. J Am Soc Nephrol 2:1078–1084, 1991.[Abstract]
  28. Kitiyakara C. C.S. Wilcox, Antioxidants for hypertension. Curr Opin Nephrol Hypertens 7:531–538, 1998.[Medline]



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