The Protective Effect Of Ginseng Polysaccharides On Ethanol-induced Behavioral Sensitization And Withdrawal, And Underlying Mechanisms

At present,most of the study on the mechanism of ethanol dependence has focused on the midbrain marginal dopamine system.In recent years,ERK / c-fos signaling pathway and hypothalamic-pituitary-adrenal axis(HPA)are increasing in the study of alcohol abuse,addiction and re-drinking.Ginseng polysaccharides(GSP)is one of the main active ingredients of ginseng.In addition,GSP has been shown to improve the learning and memory abilities of Alzheimer's disease model rats and reduce the aggressive behavior of mice and the immobility time during forced swimming.However,there have been no reports on anti-drug addiction.Therefore,the current research explored the effects of GSP on the behavior of ethanol-induced behavioral sensitization and ethanol withdrawal,and the effects of the ERK / c-fos signaling pathway and HPA axis.Objective: To investigate the effects of ginseng polysaccharides(GSP)on ethanol-induced behavioral sensitization and the effects of ginseng polysaccharides(GSP)on withdrawal behavior and plasma ACTH and Corticosterone in ethanol-dependent mice.Methods:1 Effect of ginseng polysaccharides on behavior and ERK/c-fos pathway in ethanol-induced behavioral sensitized mice120 Kunming mice were randomly divided into two parts,the first part were randomly divided into control group,GSP low-dose group(100 mg/kg),GSP middle-dose group(200 mg/kg)and GSP high-dose group(400 mg/kg),10 per group.GSP(100,200 and 400 mg/kg)were given intragastrically daily with ginseng polysaccharide solution,the control group was given the same volume of distilled water,continuously administered for 1 week,and each group were tested for locomotor activities on the 1st,3rd,and 7th days of dosing.The second part were randomly divided into five groups: control group,model group,GSP low-dose group(100 mg/kg),GSP middle-dose group(200 mg/kg)and GSP high-dose group(400 mg/kg).Except for the control group,other-groups were administered with saline or ethanol freely.The concentration of ethanol was increased gradually(3~12%,increasing by 3% every four days).After a 4-day conversion period,the initial dose of 3% ethanol was administered to establish a behavioral sensitization model.Ethanol consumption and locomotor activities of each group were measured daily during the experimental period.During the transition period,the open field test(OFT)was carried out.The activation of behavioral sensitization was determined by locomotor test during the expression period.The prefrontal cortex MDA content and the activity of SOD,CAT and GSH-Px were detected by enzyme-linked immunosorbent assay(ELISA).Western blot was used to detect the expressions of ERK,p-ERK,c-fos,NOX4,Beclin1,BAX and BCL2 protein in the prefrontal cortex.Real-time PCR was used to detect the expressions of p-ERK,c-fos mRNA in the prefrontal cortex.2 Effect of ginseng polysaccharides on withdrawal behavior and plasma ACTH and Cortisol in ethanol-dependent miceOne hundred Kunming mice were randomly divided into five groups,the control group was intraperitoneally injected with saline.The model group and GSP three dose groups(100,200 and 400 mg/kg)were intraperitoneally injected with 2.5 g/kg ethanol(20% v/v)for 15 days.The light-dark box experiment,the elevated plus maze test,and the withdrawal symptom score were performed 24 hours after the last administration of ethanol.Plasma ACTH and Corticosterone levels were measured by enzyme-linked immunosorbent assay(ELISA).Results:1 In the model of two-bottle selective drinking mode combine with behavior sensitization,GSP(100,200 and 400 mg/kg)could significantly reduce the hyperactivity caused by 9% and 12% ethanol during the transition period,and significantly inhibited the increase of locomotor activity and ethanol intake induced by 3% alcohol challenge(day25).In addition,the middle and high dose of GSP could significantly modulate the number of crawl and numbers of standing in the OFT.GSP significantly could inhibit ethanol-induced increase in MDA levels in the prefrontal cortex and increases SOD,CAT and GSH-Px activity.The Western blot results showed that GSP could downregulate the overexpression of p-ERK,c-fos and NOX4 protein in prefrontal cortex.However,ethanol and GSP had no significant effect on the expression of Beclin1,BCL2 and BAX protein in the prefrontal cortex.Besides,the real-time PCR results showed that GSP could downregulate the overexpression of p-ERK and c-fos mRNA in prefrontal cortex.In addition,the administration of GSP alone(100,200,and 400 mg / kg)had no effect on the locomotor activities of mice.2 In the ethanol dependence withdrawal model,GSP(200 and 400 mg/kg)significantly increased the number of cross and the ratio of the residence time in the bright room in the light and dark box test.The numbers of entries and time spent in the open arms were increased in GSP(200 mg/kg,400 mg/kg)-treated mice.In addition,the GSP(200 mg/kg,400 mg/kg)significantly reduced the score of withdrawal syndrome in mice compared with the model group.The ELISA results showed that GSP significantly reduced plasma Corticosterone level and increased ACTH content.Conclusion: GSP shows inhibitory effect on ethanol-induced behavioral sensitization and ethanol intake in mice.In addition,it can reduce the MDA level of the prefrontal cortex of behavior-sensitized mice,increase the activities of SOD,CAT,GSH-Px,and inhibit the overexpression of ERK,c-fos protein and mRNA.Moreover,GSP also improves the anxiety-related behavior caused by ethanol withdrawal,which may be related to modulating plasma ACH and cortisol levels.