Protective effect of Rosuvastatin on Azithromycin induced cardiotoxicity in a rat model

Abstract

Aims: Azithromycin is widely used broad spectrum antibiotic recently used in treatment protocol of COVID-19 for its antiviral and immunomodulatory effects combined with Hydroxychloroquine or alone. Rat models showed that Azithromycin produces oxidative stress, inflammation, and apoptosis of myocardial tissue. Rosuvastatin, a synthetic statin, can attenuate myocardial ischemia with antioxidant and antiapoptotic effects. This study aims to evaluate the probable protective effect of Rosuvastatin against Azithromycin induced cardiotoxicity.

Main method: Twenty adult male albino rats were divided randomly into four groups, five rats each control, Azithromycin, Rosuvastatin, and Azithromycin +Rosuvastatin groups. Azithromycin 30 mg/kg/day and Rosuvastatin 2 mg/kg/day were administrated for two weeks by an intragastric tube. Twenty-four hours after the last dose, rats were anesthetized and the following measures were carried out; Electrocardiogram, Blood samples for Biochemical analysis of lactate dehydrogenase (LDH), and creatine phosphokinase (CPK). The animals sacrificed, hearts excised, apical part processed for H&E, immunohistochemical staining, and examined by light microscope. The remaining parts of the heart were collected for assessment of Malondialdehyde (MDA) and Reduced Glutathione (GSH).

Key findings: The results revealed that Rosuvastatin significantly ameliorates ECG changes, biochemical, and Oxidative stress markers alterations of Azithromycin. Histological evaluation from Azithromycin group showed marked areas of degeneration, myofibers disorganization, inflammatory infiltrate, and hemorrhage. Immunohistochemical evaluation showed significant increase in both Caspase 3 and Tumor necrosis factor (TNF) immune stain. Rosuvastatin treated group showed restoration of the cardiac muscle fibers in H&E and Immunohistochemical results.

Significance: We concluded that Rosuvastatin significantly ameliorates the toxic changes of Azithromycin on the heart.

Keywords: Azithromycin; Heart; Rat; Rosuvastatin.

Fig. 1

Summary of the used experimental methods and results.

Fig. 2

Changes in electrocardiogram among groups: C: control, Az: Azithromycin treated group, R: Rosuvastatin and Az + R: Azithromycin + Rosuvastatin.

Fig. 3

Distribution of heart rhythm among group: C: control, Az: Azithromycin treated group, R: Rosuvastatin and Az + R: Azithromycin + Rosuvastatin.

Fig. 4

Comparisons of Electrocardiogram parameters between study groups (N = 20). *Statistically significant at p < 0.05. A: HR; B: QTc; C: PR.

Fig. 5

Comparisons of biochemical and oxidative stress markers between study groups (N = 20). *Statistically significant at p < 0.05. A: LDH; B: CPK; C: MDA; D: GSH.

Fig. 6

Photomicrograph of the heart ventricle (A & B): Control group showing normal cardiac muscle fibers with acidophilic cytoplasm and oval, large pale nuclei (N). Some cardiomyocytes are binuclear (BI). Myofibers are arranged in bundles with narrow spaces in between (S) and connected with intercalated discs (arrows). (C & D): Azithromycin group showing areas of severe degeneration in cardiac muscle fibers (arrows), pyknotic nuclei (PN), distortion of the typical arrangement of myofibers with massive wide spaces (S), and hemorrhage (Hge). (E): Azithromycin + Rosuvastatin group showing the restored arrangement of cardiac muscle fibers with narrow spaces in between (S). Ventricle cavity (CA). A, C, D, E (H&E; ×400) and B (H&E; ×1000).

Fig. 7

Photomicrograph of the heart ventricle stained with caspase 3 A: control group is showing few positive cytoplasmic immune reactions. B: Azithromycin treated group showing strong positive immune reactions C: Azithromycin and Rosuvastatin treated group shows few positive immune reactions. (caspase 3; ×400).

Fig. 8

Photomicrograph of the heart ventricle stained with TNF α A: control group showing few positive cytoplasmic immune reaction B: Azithromycin treated group showing strong positive immune reactions for TNF α. C: Azithromycin and Rosuvastatin treated group showing few positive immune reactions (TNF α; ×400).

Fig. 9

Comparisons of immune-histochemical markers between the study groups (N = 20). *Statistically significant at p < 0.05. A: TNF; B: Caspase 3.