Hesperidin ameliorates immobilization-stress-induced behavioral and biochemical alterations and mitochondrial dysfunction in mice by modulating nitrergic pathway

Abstract

The present study was aimed to evaluate the protective effect of hesperidin against immobilization-stress-induced alterations in biochemical, behavioral, and mitochondrial functions in mice. In many instances neuroscientists have reported that acute immobilization stress for 6 h resulted in anxiety and impaired locomotor activity due to excess oxidative-nitrergic stress, depletion of antioxidant defense mechanisms, and mitochondrial dysfunction in animals. In the present study, 6 h of acute immobilization stress had significantly altered the behavioral (anxiety and memory) and biochemical parameters coupled with mitochondrial dysfunction in Swiss albino mice. Fourteen days of pretreatment with Hesperidin (50 and 100 mg/kg, p.o.) significantly and dose-dependently inhibited the behavioral and biochemical alterations and mitochondrial dysfunction caused by acute immobilization stress. Furthermore, pre-treatment of l-arginine (50 mg/kg, i.p.), a nitric oxide precursor, reversed the protective effect of Hesperidin (50 and 100 mg/kg) (P < 0.05). In contrast, pretreatment of l-NAME (5 mg/kg, i.p.), a nitric oxide synthase inhibitor, potentiated the protective effect of Hesperidin (P < 0.05). These results suggest the possible involvement of nitrergic pathway in the protective effect Hesperidin against immobilization-stress-induced behavioral, biochemical, and mitochondrial dysfunction in mice.

Figure 1

Illustration of the protocol used for the study.

Figure 2

Effect of Hesperidin on immobilization-stress-induced altered locomotor activity. IS: immobilization stress, Hes: Hesperidin, and pr: protein, all the values are expressed as mean ± SEM. ^aP < 0.05 compared to naïve, ^bP < 0.05 compared to control (IS), ^cP < 0.001 compared to Hesperidin (50), ^dP < 0.05 compared to Hesperidin-100, and ^eP < 0.05 compared to l-arginine (50).

Figure 3

Effect of Hesperidin on immobilization-stress-induced anxiety-like behavior in elevated plus maze test (time spent in open arm). IS: immobilization stress, Hes: Hesperidin, and pr: protein, all the values are expressed as mean ± SEM. ^aP < 0.05 compared to naïve, ^bP < 0.05 compared to control (IS), ^cP < 0.001 compared to Hesperidin (50), ^dP < 0.05 compared to Hesperidin-100, and ^eP < 0.05 compared to l-arginine (50).

Figure 4

Effect of Hesperidin on immobilization-stress-induced anxiety-like behavior in Elevated plus maze test (No. of entries in to open arm). IS: immobilization stress, Hes: Hesperidin, and pr: protein, all the values are expressed as mean ± SEM. ^aP < 0.05 compared to naïve, ^bP < 0.05 compared to control (IS), ^cP < 0.001 compared to Hesperidin (50), ^dP < 0.05 compared to Hesperidin-100, and ^eP < 0.05 compared to l-arginine (50).

Figure 5

Effect of Hesperidin on immobilisation stress induced anxiety-like behavior in mirror chamber test. IS: immobilization stress, Hes: Hesperidin, and pr: protein, all the values are expressed as mean ± SEM. ^aP < 0.05 compared to naïve, ^bP < 0.05 compared to control (IS), ^cP < 0.001 compared to Hesperidin (50), ^dP < 0.05 compared to Hesperidin-100, and ^eP < 0.05 compared to l-arginine (50).

Figure 6

Effect of naringin on immobilisation stress induced anxiety-like behaviour in mirror chamber test (No. of entries). IS: immobilization stress, Hes: Hesperidin, and pr: protein, all the values are expressed as mean ± SEM. ^aP < 0.05 compared to naïve, ^bP < 0.05 compared to control (IS), ^cP < 0.001 compared to Hesperidin (50), ^dP < 0.05 compared to Hesperidin-100, and ^eP < 0.05 compared to l-arginine (50).

Figure 7

Effect of Hesperidin on immobilization stress induced mitochondrial dysfunction in mice (% naïve). IS: immobilization stress, Hes: Hesperidin, and pr: protein, all the values are expressed as mean ± SEM (% of naïve group), ^aP < 0.05 compared to naïve, ^bP < 0.05 compared to control (IS), ^cP < 0.001 compared to Hesperidin (50), ^dP < 0.05 compared to Hesperidin-100, and ^eP < 0.05 compared to l-arginine (50).