Figure 1.
Schematic representation of several groups used in the study showing the treatments received by the diabetic rats for 14 days.
On day 15th these rats were subjected to the LAD coronary artery ligation followed by a one hour reperfusion induced myocardial injury. Moreover, these groups were assessed for several biochemical parameters, histological evaluations, ultrastructural studies, myocardial apoptosis markers and western blot analysis.
Figure 2.
Effect of hesperidin and GW9662 on mean arterial pressure (MAP) and left ventricular (LV) function following ischaemia/reperfusion (I/R) in diabetic rats.
(A) MAP, (B) left ventricular end-diastolic pressure (LVEDP), (C) maximal positive rate of left ventricular pressure (+LVdP/dtmax) and (D) maximal negative rate of −LVdP/dtmax. Data are expressed as the mean ± standard error (n = 22/group). Significance was determined by repeated measures analysis of variance followed by the Bonferroni's post hoc test: *p<0.05, **p<0.001 vs Diabetic SHAM; #p<0.05, ##p <0.001 vs Diabetic I/R; $p<0.01 vs respective Hesperidin.
Table 1.
Effect of hesperidin and GW9662 on cardiac injury marker enzymes, lipid peroxidation, TNF-α and antioxidant parameters.
Table 2.
Mean area at risk and infarct area in rats in the different experimental groups.
Figure 3.
Photomicrograph showing the histopathological changes in the rat myocardium.
(A) represents haematoxylin and eosin staining [(H&E) ×200] in diabetic sham group, B diabetic ischaemia/reperfusion (I/R) group (H&E ×200), (C) diabetic I/R + hesperidin treatment (100 mg/kg) (H&E ×200), (D) diabetic I/R + GW9662 (1 mg/kg) (H&E ×200) and (E) diabetic I/R rats + GW9662 + hesperidin (H&E ×200).
Table 3.
Histopathological changes in all experimental groups.
Figure 4.
Photomicrograph showing the ultrastructural changes in the rat myocardium.
(A) represents transmission electron microscope [(TEM) ×4800] in diabetic sham group, B diabetic ischaemia/reperfusion (I/R) group (TEM ×3500), (C) diabetic I/R + hesperidin treatment (100 mg/kg) (TEM ×3500), (D) diabetic I/R + GW9662 (1 mg/kg) (TEM ×3500) and (E) diabetic I/R rats + GW9662 + hesperidin (TEM ×3500).
Figure 5.
Photomicrograph of myocardial tissue sections showing Bcl-2 (A–E), Bax proteins expression (F–J) and TUNEL-positive cells (K–O).
Apoptotic nuclei are indicated by arrows in panels (×400). (A, F and K) represents Diabetic sham rats treated with vehicles, (B, G and L) diabetic ischaemia/reperfusion (I/R) rats treated with vehicles, (C, H and M) diabetic I/R rats treated with hesperidin, (D, I and N) diabetic I/R rats treated with GW9662 and (E, J and O) diabetic I/R rats treated with GW9662 and hesperidin. (P) represents the quantitative analysis of the Bcl-2, Bax proteins and TUNEL-positive cells in the different groups. Data are expressed as the mean ± standard error (n = 6). *p<0.05, *p<0.001 vs diabetic SHAM; #p<0.05, ##p<0.001 vs Diabetic I/R group; @p<0.01 vs Diabetic I/R + hesperidin.
Figure 6.
Effect of hesperidin and GW9662 (PPAR-γ antagonist) on PPAR-γ expression (top) in ischaemia/reperfusion (I/R)-induced myocardial infarction in diabetic rats.
The bar graphs show PPAR-γ protein expression that has been normalized to β-actin (bottom). Data are expressed as a ration of the value determined for the vehicle (diabetic sham) (set as 100%). Data are expressed as the mean ± standard error. *p<0.05 vs Diabetic SHAM; #p<0.01, #p<0.001 vs Diabetic I/R; @p<0.05 vs Hesperidin.