Differential effect of heme oxygenase-1 in endothelial and smooth muscle cell cycle progression

doi:10.1016/S0006-291X(02)02054-5

Biochemical and Biophysical Research Communications

Volume 296, Issue 5, 6 September 2002, Pages 1077-1082

https://doi.org/10.1016/S0006-291X(02)02054-5 Get rights and content

Abstract

Arterial remodeling in response to pathological insult is a complex process that depends in part on the balance between vascular cell apoptosis and proliferation. Studies in experimental models suggest that HO-1 mediates neointimal formation while limiting lumen stenosing, indicating a differential effect on vascular endothelial (EC) and smooth muscle cells (SMC). We investigated the effect of HO-1 expression on cell cycle progression in EC and SMC. The addition of SnMP (10 μM), an inhibitor of HO activity, to EC or SMC for 24 h, resulted in significant abnormalities in DNA distribution and cell cycle progression compared to cells treated with the HO-1 inducers, heme (10 μM) or SnCl₂ (10 μM). SnMP increased G₁ phase and decreased S and G₂/M phases in EC while heme or SnCl₂ decreased G₁ phase, but increased S and G₂/M phases (p<0.05). Opposite effects were obtained in SMC. SnMP decreased G₁ phase and increased S and G₂/M phases while heme or SnCl₂ increased G₁ phase but decreased S and G₂/M phases (p<0.05). Our data demonstrate that HO-1 regulates the cell cycle in a cell-specific manner; it increases EC but decreases SMC cycle progression. The mechanisms underlying the HO-1 cell-specific effect on cell cycle progression within the vascular wall are yet to be explored. Nevertheless, these findings suggest that cell-specific targeting of HO-1 expression may provide a novel therapeutic strategy for the treatment of cardiovascular diseases.

Section snippets

Materials and methods

Cell culture conditions. Human dermal microvessel endothelial cells (EC) were a gift from Dr. Michael Dillon (National Center for Infectious Diseases, Atlanta, GA) and grown in MCDB131 medium (Gibco-BRL, Grand Island, NY) supplemented with 10% fetal bovine serum (FBS), 10 ng/ml endothelial growth factor (Sigma, St. Louis, MO), and 1 μg/ml hydrocortisone (Sigma). Human SMC were obtained from ATCC (Manassas, VA) and grown in Dulbecco's modified Eagle's medium (DMEM) with 10% FBS. The cells were

Differential effect of inducers and inhibitors on HO-1 protein

EC and SMC were examined for the levels of HO-1 and HO-2 proteins by Western blot analysis. The results of three representative experiments are shown in Fig. 1. Both EC (Fig. 1A) and SMC (Fig. 1B) demonstrated low basal levels of HO-1 protein and a several fold increase after treatment with heme (lane 2) or SnCl₂ (lane 3), as compared with control cells (lane 1). Treatment with heme or SnCl₂ did not significantly modulate HO-2 protein in either cell type.The addition of SnMP (lane 4), a known

Discussion

This study demonstrates, for the first time, that the heme–HO system participates in the regulation of cell proliferation in EC and SMC in a cell-specific manner. Two key findings substantiate this conclusion. The first key finding is that heme and SnCl₂, which are known to induce HO activity in tissues [32], [33] and other mammalian cells [34], act as inducers of HO-1 gene expression in both EC and SMC. Upregulation of HO-1 gene expression in SMC was associated with a decrease in cell cycle

Acknowledgements

This work was supported by AHA Grant 50948T and NIH Grants HL55601 and HL34300. We thank Ms. Sylvia Shenouda for her technical assistance and Ms. Jennifer Brown for her excellent secretarial assistance.

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Differential effect of heme oxygenase-1 in endothelial and smooth muscle cell cycle progression

Abstract

Section snippets

Materials and methods

Differential effect of inducers and inhibitors on HO-1 protein

Discussion

Acknowledgements

Int. J. Biochem.

Arch. Biochem. Biophys.

Kidney Int.

Kidney Int.

J. Biol. Chem.

Biochem. Biophys. Res. Commun.

Immunol. Today

Am. J. Med. Sci.

Kidney Int.

J. Lab. Clin. Med.

The biological significance and physiological role of heme oxygenase

Cell. Physiol. Biochem.

Regulation of ferritin and heme oxygenase synthesis in rat fibroblasts by different forms of iron

Proc. Natl. Acad. Sci. USA

Functional analysis of cDNAs for two types of human heme oxygenase and evidence for their separate regulation

J. Biochem. Tokyo

Gene transfer of human heme oxygenase into coronary endothelial cells potentially promotes angiogenesis

J. Cell. Biochem.

Differential effects of heme oxygenase isoforms on heme mediation of endothelial intracellular adhesion molecule 1 expression

J. Pharmacol. Exp. Ther.

Transfection of the human heme oxygenase gene into rabbit coronary microvessel endothelial cells: protective effect against heme and hemoglobin toxicity

Proc. Natl. Acad. Sci. USA

Oxidative stress causes enhanced endothelial cell injury in human heme oxygenase-1 deficiency

J. Clin. Invest.

Endothelial-cell heme uptake from heme proteins: induction of sensitization and desensitization to oxidant damage

Proc. Natl. Acad. Sci. USA

Induction of heart heme oxygenase-1 (HSP32) by hyperthermia: possible role in stress-mediated elevation of cyclic 3′:5′-guanosine monophosphate

J. Pharmacol. Exp. Ther.

Induction of heme oxygenase-1 inhibits the monocyte transmigration induced by mildly oxidized LDL

J. Clin. Invest.

Evidence for cytokine-inducible nitric oxide synthesis from l-arginine in patients, receiving interleukin-2 therapy

J. Clin. Invest.

Induction of heart heme oxygenase-1 (HSP32) by hyperthermia: possible role in stress-mediated elevation of cyclic 3′:5^′-guanosine monophosphate