Effects of gallic Acid and cyclosporine a on antioxidant capacity and cardiac markers of rat isolated heart after ischemia/reperfusion

Abstract

Background: Myocardial infarction is one of the important causes of death during old ages. Gallic acid as an antioxidant or cyclosporine A (CsA) as inhibitor of mitochondrial permeability transition pore (mPTP) alone could prevent these complications to some extent, but their combination effect has not been investigated.

Objectives: The aim of this study was to determine the combined effect of gallic acid and CsA on antioxidant capacity of isolated heart tissues during ischemia reperfusion.

Materials and methods: EIGHTY MALE WISTAR RATS WERE RANDOMLY ASSIGNED TO DIFFERENT GROUPS: sham, control (Ca, received saline, 1 mL/kg); 3 groups were pretreated with gallic acid (G1a: 7.5, G2a: 15, G3a: 30 mg/kg) for 10 days, and the other 3 groups were pretreated with gallic acid and received CsA (0.2 µM) for 10 minutes before induction of ischemia and during the first 10 minutes of reperfusion (G1b, G2b and G3b) and the last group received CsA alone (Cb). After 10 days of pretreatment, the heart was isolated and transferred to the Langendorff apparatus and exposed to 30 minutes ischemia followed by 60 minutes of reperfusion. After that cardiac markers and antioxidant enzymes were assessed in cardiac tissues.

Results: Lactate dehydrogenase (LDH), Superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX) activity increased and malondialdehyde (MDA) decreased in animals pretreated with gallic acid significantly. However, pretreatment with gallic acid followed by CsA during reperfusion improved the antioxidant capacity and cardiac marker enzymes and restored the lipid peroxidation more effective than gallic acid or CsA alone. Nevertheless, CsA did not change the cardiac marker enzymes significantly.

Conclusions: Gallic acid and CsA combination improved antioxidant capacity and cell membrane integrity more than each one alone. Therefore, it can be a therapeutic approach to reduce the I/R injury.

Keywords: Antioxidant Capacity; Cyclosporine A; Gallic Acid; Ischemia Reperfusion; Myocardial Infarction.

Figure 1.. TBARS levels in Heart of Rats After 30 Minutes Ischemia and 60 Minutes Reperfusion (Mean ± SEM, n = at least 8)

The animals received three different doses of gallic acid (G7.5, G15 and G30 mg/kg) for 10 days before the beginning of I/R studies. Cyclosporine A (0.2 µM) was administered at the onset of reperfusion (CsA). Control animals received normal saline as the solvent of gallic acid. Sham animals were not exposed to I/R. * indicates significant difference with control. + indicates significant difference with sham. (+ or * P < 0.05, ** or ++ P < 0.01, *** or +++, P < 0.001, one-way ANOVA followed by LSD test).

Figure 2.. CAT Activity in Heart of Rats After 30 Minutes Ischemia and 60 Minutes of Reperfusion (Mean ± SEM, n = at least 8)

The animals received three different doses of gallic acid (G7.5, G15 and G30 mg/kg) for 10 days before the beginning of I/R studies. Cyclosporine A (0.2 µM) was administered at the onset of reperfusion (CsA). Control animals received normal saline as the solvent of gallic acid. Sham animals were not exposed to I/R. * indicates significant difference with control. (* P < 0.05, ** P < 0.01, one-way ANOVA followed by LSD test).

Figure 3.. SOD activity in Heart of Rats After 30 Minutes Ischemia and 60 Minutes of Reperfusion (Mean ± SEM, n = at least 8)

The animals received three different doses of gallic acid (G7.5, G15 and G30 mg/kg) for 10 days before the beginning of I/R studies. Animals received cyclosporine A (0.2 µM) at the onset of reperfusion (CsA); Control animals received normal saline as the solvent of gallic acid. Sham animals were not exposed to I/R.* indicates significant difference with control. + indicates significant difference with sham. (* or + P < 0.05, ** or ++ P < 0.01, +++ P < 0.001, one-way ANOVA followed by LSD test). a = compared with CsA, b = compared with each corresponding dose of gallic acid alone.

Figure 4.. GPX Activity in Heart of Rats After 30 Minutes Ischemia and 60 Minutes of Reperfusion (Mean ± SEM, n = at least 8)

The animals received three different doses of gallic acid (G7.5, G15 and G30 mg/kg) for 10 days before the beginning of I/R studies. Animals received cyclosporine A (0.2 µM) at the onset of reperfusion (CsA); Control animals received normal saline as the solvent of gallic acid. Sham animals were not exposed to I/R.* indicates significant difference with control.* indicates significant difference with sham. (P < 0.05, ** P < 0.01, *** P < 0.001, one-way ANOVA followed by LSD test). a = compared with CsA, b = compared with each corresponding dose of gallic acid alone.

Figure 5.. TAS Activity in Heart of Rats After 30 Minutes Ischemia and 60 Minutes of Reperfusion (Mean ± SEM, n = at least 8)

The animals received three different doses of gallic acid (G7.5, G15 and G30 mg/kg) for 10 days before the beginning of I/R studies. Animals received cyclosporine A (0.2 µM) at the onset of reperfusion (CsA), Control animals received normal saline as the solvent of gallic acid. Sham animals were not exposed to I/R.* indicates significant difference with control. + indicates significant difference with sham. (P < 0.05, ++ P < 0.01, +++ P < 0.001, one-way ANOVA followed by LSD test). a = compared with CsA, b = compared with each corresponding dose of gallic acid alone.