The Pharmacological Validation Of Qi-Deficiency Syndrome With Chinese Herbal Medicine In A Depression Model And An Associated Molecular Biology Study In Balb/c Strain Mice

The differentiation treatment based on heterogeneity of disease is not conducted,which may be one of the reasons that treatment of modern medicine on depression is not satisfying.Traditional Chinese Medicine(TCM)is personalized medicine that advocates individual treatment and uses right prescriptions for the syndromes of disease clinically.Previously we have found that Balb/c mice demonstrated strain-dependent Qi-deficiency like syndromes in the same disease models.This study focuses on the Chinese herbal formula verification of Qi-deficiency syndrome of CUMS(chronic unpredictable mild stress)-induced depression in Balb/c mice and explores the molecular mechanism of the drug in the Qi-deficiency susceptibility model of depression.Objective:1.This study focused on the formula verification of the the TCM Qi-deficiency syndrome and depression-like behavior in Balb/c and 129/S1 mice following CUMS.2.To verify whether depression syndromes of B/c female mice is as B/c male mice.3.To detect the antidepressant efficiency of Sijunzi Decoction(SJZD)pretreatment.Methods:Corresponding to the objective,the experiment includes three parts:1.Male Balb/c and 129/S1 mice were randomly assigned into two groups,the control group which were not given stress and the model group which were subjected to 4 weeks CUMS to induce depression.We assessed the behaviors of depression in the mice by sucrose preference test(SPT)and forced swim test(FST).We assessed the Qi-deficiency phenotype by grip strength and exhausted swim test.After the four-week stressor,the model group was randomly divided into four groups,the vehicle(Veh)group,Yueju(YJ)group,Sijunzi Decoction(SJZD)group and Fluoxetine(FLX)group.Control(CTL)group and vehicle group were administrated saline.YJ group was administrated Yueju pill via intragastric administration,with the concentration of 0.2g/mL.SJZD group was administrated Sijunzi Decoction via intragastric administration,with the concentration of 0.5g/mL.Fluoxetine group was administrated Fluoxetine,with the concentration of 2mg/mL intraperitoneally.Drug treatment lasted for one week.After one-week drug administration,animal behavioral tests were tested,including Open field test(OFT),Sucrose preference test(SPT),Forced swim test(FST),Tail suspension test(TST)and Novelty suppressed feeding test(NSF),to assess the antidepressant-like effect and the improvements on Qi-dificiency syndrome such as grip strength,exhausted swim test and so on.When All behavioral tests finished,western blotting were used to detect the protein expression of hippocampus and muscle of PKA and p-CREB to study the molecular mechanisms.2.4-week CUMS was used to induce depression on the female Balb/c mice.Sijunzi Decoction was administrated to mice and the syndromes were verified based on pharmacological effect.We assessed the behaviors of depression and the Qi-deficiency phenotype in the mice.The changes of PKA-CREB signaling in muscle and hippocampus of mice were observed via Western Blotting.3.Male Balb/c mice were subjected to 3-week CUMS to induce depression.We assessed the behaviors of depression and Qi-deficiency phenotype.Then the model group was randomly divided into four groups,the vehicle(Veh)group,Yueju(YJ)group,Sijunzi Decoction(SJZD)group and Fluoxetine(FLX)group.Control(CTL)group and vehicle group were administrated saline.YJ group was administrated Yueju pill via intragastric administration,with the concentration of 0.2g/mL.SJZD group was administrated Sijunzi Decoction via intragastric administration,with the concentration of 0.5g/mL.Fluoxetine group was administrated intraperitoneally,with the concentration of 2mg/mL.Drug treatment lasted for one week.After one-week drug administration,all animal behavioral tests were tested to assess the pretreatment of SJZD.Results:1.Both B/c and 129S mice performed similar depression-like behaviors in SPT and FST,but only Balb/c mice performed Qi-deficiency pattern in grip strength,loaded swim time and degree of redness following 4 week CUMS.Qi-tonification treatment reliefed Qi deficiency phenotype and outperformed Qi-unobstruction treatment in alleviation of depression-like behaviors in Balb/c mice,while Qi-unobstruction but not Qi-tonification treatment alleviates depression-like behaviors in 129/S1 mice.SJZD’s treatment efficay associated reversal of deficient PKA-CREB signaling in muscle and hippocampus of Balb/c mice is better than YJ.Meantime,YJ reversed deficient PKA-CREB signaling in muscle and hippocampus of 129/S1 mice while SJZD could not.2,SJZD improved Qi-deficiency like syndromes of Balb/c female mice by reversing deficits in grip strength,exhausted swim test.SJZD also reversed depressive like behaviors by increasing the sucrose preference and so on.The decreased phosphorylation of the CREB(p-CREB)and PKA expression of hippocampus and muscle in CUMS mice were reversed by SJZD.3.SJZD improved depressive like behaviors by reversing deficits in the sucrose preference,immobility and latency,as well as Qi-deficiency like syndromes in grip strength and degree of redness,while YJ failed to exert these effects.There is no statistical difference between the control group and the control+SJZD group in SPT,FST,TST,grip strength,loaded swim time and degree of redness.Conclusion:1.SJZD alleviates depression-like behaviors and Qi-deficiency phenotype induced by CUMS in Balb/c mice.Improving PKA-CREB signaling in periphery and brain may be an essential mechanism.2.Sex does not make differences of Qi-deficiency syndrome model in depression in the same strain.3.Pretreatment of Qi-tonification prevented anhedonic response in Balb/c mice following three-week CUMS.4.No antidepressant effect of Qi-tonification treatment in non-stress Balb/c mice.