Cardiovascular pharmacology
Rosuvastatin promotes angiogenesis and reverses isoproterenol-induced acute myocardial infarction in rats: Role of iNOS and VEGF

https://doi.org/10.1016/j.ejphar.2012.06.022Get rights and content

Abstract

Several reports highlighted the cardioprotective effect of statins after different types of ischemic injury. We studied the effect of rosuvastatin on acute myocardial infarction induced experimentally in rats focusing on angiogenesis as a potential mechanism underlying the drug effect. Acute myocardial infarction was induced by injecting the rats with two doses of isoproterenol (85 mg/kg/24 h, s.c.). Rats were examined for their electrocardiographic pattern and myocardial fibrosis one week after injection of isoproterenol (time for initiating therapy) and eight weeks thereafter (the end of therapeutic period) to examine the progression of the injury. Examination of the heart tissues at the end of week 9 showed a non significant decrease in the degree of myocardial fibrosis compared to those observed at week 1, indicating a slow rate of recovery from isoproterenol-induced injury. Treatment with rosuvastatin (5 or 10 mg/kg) for 8 weeks in myocardial-infarct rats enhanced the electrocardiographic pattern, reduced serum cardiac biomarkers, reduced tissue tumor necrosis factor-α (TNF-α) and upregulated vascular endothelial growth factor (VEGF) level. In addition, immunohistochemical staining revealed higher expression of inducible nitric oxide synthase (iNOS), VEGF and CD34 (a marker for microvessel density) in the cardiac tissues after treatment with rosuvastatin compared to control group. The immunostaining for VEGF was positively correlated with microvessel density and iNOS. Overall, the current results provide evidence that the effect of rosuvastatin on myocardial-infarct rats involves induction of angiogenesis.

Introduction

Cardiovascular diseases account for high number of deaths all over the world. Myocardial infarction is the common presentation of cardiovascular disease (Patel et al., 2010). After myocardial ischemia, the heart cells die and collagen deposition ensues in its place (Krijnen et al., 2002). Angiogenesis is defined as the formation of new blood vessels from the preexisting ones. Angiogenesis may be beneficial in some pathologic conditions, such as in ischemic diseases (Iglarz et al., 2001). Ischemia-induced angiogenesis takes place as a compensatory mechanism to provide blood supply to ischemic tissues and preserves blood flow in ischemic heart or leg (Heil and Schaper, 2004).

Therapeutic angiogenesis is a new strategy that has been emerged to treat tissue ischemia by promoting the proliferation of collateral vessels. This strategy received great attention as a promising therapy developed in ischemic diseases (Freedman and Isner, 2001). In ischemic tissues, hypoxia-induced angiogenesis takes place due to the increase in expression of some important growth factors, such as vascular endothelial growth factor (VEGF) (Lee et al., 2000). Indeed, NO has been reported to be an important mediator in the cardiovascular system and is thought to be critical in signal transduction through the ischemic myocardium (Xi et al., 1999). Nitric oxide is generated by nitric oxide synthase (NOS) enzyme. Inducible nitric oxide synthase (iNOS) isoform has been reported to play an important role in reducing the infarct size during myocardial infarction (Shah and Wainwright, 2000). It has been demonstrated that iNOS and VEGF are over expressed and promote tumor angiogenesis and metastasis (Chen et al., 2009). In addition, iNOS-derived NO was reported to promote survival of ischemic tissue by stimulating angiogenesis (Kane et al., 2001).

Some clinical trials established the benefits of statins on coronary heart diseases and strokes (Heart Protection Study Collaborative Group, 2003, Law et al., 2003), ischemic cerebrovascular events (Greisenegger et al., 2004) and hind limb ischemic tissues (Sata et al., 2001). Statins were reported to improve the endothelial function by mechanisms that are unrelated to the cholesterol lowering effect. Some of these mechanisms involve prevention of inflammation, down regulation of the systemic inflammatory response (Liu et al., 2009) and induction of angiogenesis (Hamelin and Turgeon, 1998).

Based on the previous demonstration for the cardio protective role of statins in ischemic myocardium, this study investigated one putative mechanism that might contribute to this beneficial effect. Focusing on the angiogenic role, the present study was designed to examine the pro-angiogenic effect of rosuvastatin in myocardial infarct rats.

Section snippets

Experimental animals

Male adult Wistar rats (200–250 g) were used in the present study. Rats were maintained in groups of five in stainless steel cages under controlled laboratory conditions (23–28 °C and normal dark/light cycle). Rats were acclimatized to the experiment conditions for seven days before starting the experiment. Water and food were provided ad libitum. All animal care and experimental procedures were approved by the Institutional Laboratory Animal Care and Use Committee at Faculty of Pharmacy, Suez

Effect of treatment with rosuvastatin on percentage survival and ECG pattern

In the present study, isoproterenol (85 mg/kg/24 h, s.c.) was administered to induce acute myocardial infarction in rats. Injection of isoproterenol resulted in a decrease in the percentage of surviving rats (75%) after the first week of injection compared to (62.5%) after 9 weeks of injection versus a surviving percentage equals 100% in saline-treated rats. Treatment with rosuvastatin did not significantly modify the number of surviving rats compared to isoproterenol (week 9) group (Table 1).

Discussion

The current study was the first to investigate the pro-angiogenic effect of rosuvastatin in experimentally-induced acute myocardial infarction in rats. In the present study, acute injection with isoproterenol induced myocardial fibrosis and necrosis. Evaluation of myocardial fibrosis at week 9 highlighted a non significant decrease in fibrosis grades compared to those recorded at the end of week 1, indicating a slow rate of recovery from damage induced by isoproterenol. The present results

Sources of support

The authors did not receive a fund from any organization.

Acknowledgments

The authors wish to acknowledge the generous gift of rosuvastatin from Al-Hekma Pharmaceutical Co. (6th of October City, Egypt).

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