4.1 Effect of rutin or fluoxetine on various alcohol withdrawal syndrome
The development of alcohol withdrawal symptoms was assessed on day 22 by scoring withdrawal-induced physical signs of craving showing depressive like behaviour at 2, 4, 6, and 8 hours. Rats in the alcohol control group exhibited significant (t = 4.24; F = 4.53; P < 0.0075) higher withdrawal score compared with the normal control group (Fig. 1A). However, prior treatment of rutin 100 and 200 mg/kg did exhibit non-significant reduction of withdrawal symptoms compared to alcohol control rats (Fig. 1A). In addition, after treatment with rutin 200 mg/kg and fluoxetine induced significant ( t = 3.54; F = 12.61; P < 0.0001) attenuation withdrawal symptoms compared to alcohol control (Fig. 1B).
4.2. Effects of rutin or fluoxetine on immobility time by FST in alcohol withdrawal induced depressive like behaviour in rats
Our experimental indicated that, immobility time was found to significantly increased in vehicle treated alcohol withdrawal rats (t = 10.07, F = 48.26, p < 0.0001) in FST on day 30th day. Post hoc analysis demonstrated that immobility time was significantly attenuated with fluoxetine and rutin treatment for 30 days in FST. The attenuation with rutin treatment was found to be in dose dependent manner as shown in Fig. 3.
4.3. Effects of rutin or fluoxetine on immobility time by TST in alcohol withdrawal induced depressive like behaviour in rats
In the present experimental findings by one way ANOVA suggested that immobility time by TST was also found to elevated significantly (t = 18.04, F = 116.6, p < 0.0001) due alcohol withdrawal on 30th day. Post hoc analysis indicated that, fluoxetine or rutin significantly attenuated immobility time in TST which was similar to that of TST. The attenuation of immobility time with rutin treatment was found to be dose dependent manner (Fig. 4).
4.4. Effects of rutin or fluoxetine on serum corticosterone levels in alcohol withdrawal induced depressive like behaviour in rats
One way ANOVA analysis demonstrated that, serum corticosterone levels was found to increased significantly (t = 5.642, F = 25.59, p < 0.0001) in alcohol withdrawal group. Post hoc results showed that rats in vehicle treated alcohol withdrawal group elicited increased serum corticosterone which were subsequently attenuated significantly with rutin 200 mg/kg (Fig. 4).
4.5. Effects of rutin or fluoxetine on MDA formation in alcohol withdrawal induced depressive like behaviour in rats
In the present experimental findings by one way ANOVA revealed elevation of MDA formation in significant (t = 5.97, F = 13.29, p < 0.0001) manner in alcohol withdrawal rats on 30th day. Post hoc analysis suggested that, fluoxetine or rutin significantly prevented MDA formation due to alcohol withdrawal rats. The prevention of MDA formation with rutin treatment was observed at 200 mg/kg dose level per se (Fig. 5).
4.6. Effects of rutin or fluoxetine on NO levels in alcohol withdrawal induced depressive like behaviour in rats
Our experimental findings by one way ANOVA suggested that NO levels in brain homogenate was increased significantly (t = 18.40, F = 160.62, p < 0.0001) in alcohol withdrawal rats on 30th day. Post hoc analysis revealed that, rutin treatment elicited significant attenuation of elevated NO levels in brain homogenate. The degree of attenuation was found to highest and significant with rutin 200 mg/kg and fluoxetine treatment. Rutin 50 and 100 mg/kg exhibited non-significant attenuation effects on NO levels in alcohol withdrawal induce depressive like behaviour in rats (Fig. 6).
4.7. Effects of rutin or fluoxetine on SOD activity in alcohol withdrawal induced depressive like behaviour in rats
Our findings by one way ANOVA suggested that SOD activity was declined significantly (t = 16.90, F = 104.30, p < 0.0001) alcohol withdrawal group. Post hoc analysis revealed that, rutin or fluoxetine treatment elicited significant amelioration of SOD activity. The rutin treatment showed improvement in SOD activity in dose dependent manner due to alcohol withdrawal (Fig. 7).
4.8. Effects of rutin or fluoxetine on CAT activity in alcohol withdrawal induced depressive like behaviour in rats
One way ANOVA indicated that CAT activity was found to declined significantly (t = 11.51, F = 52.14, p < 0.0001) alcohol withdrawal group. Post hoc analysis suggested that rutin or fluoxetine treatment significantly prevented decrement in CAT activity due to alcohol withdrawal. Rutin treatment exhibited dose dependent decrement in CAT activity due to alcohol withdrawal (Fig. 8).
4.9 Effects of rutin or fluoxetine on GSH levels alcohol withdrawal induced depressive like behaviour in rats
Figure 9 presented NPSH contents were estimated as GSH levels and one way ANOVA showed a significant reduction(t = 10.19, F = 5.16, p < 0.0001) in the level of NPSH contents in the alcohol withdrawal group. Rutin treatment significantly ameliorated the NPSH contents in dose dependent manner due to alcohol withdrawal. Furthermore, fluoxetine a standard antidepressant exhibited non-significant effects on NPSH contents (Fig. 9)
4.10. Histopathological studies
Histopathological findings showed that, alcohol withdrawal group elicited oedematous and inflammatory neuronal infiltration and impaired structural alterations. Treatment with of rutin 200 mg/kg exhibited normal neuronal with minor swelling and inflammatory changes due to alcohol withdrawal (Fig. 10).