| Rationales:Depression is a common mood disorder disease,which always accompanied with a variety of psychological symptoms and physical symptom group,and the incidence of depression increased year by year.Depression has a serious harmfulness.According to the study of the global burden of diseases,the loss of healthy life caused by depression is in the second place in all diseases,and patients with major depression would have suicidal thoughts and behavior.Drugs used to treat depression on the market at present were given priority to with western medicine.But there is low efficient,more side effects and narrow function spectrum in these drugs,which has the single acting way without treatment effect to the accompanying symptoms of depression.In recent years,domestic researchers will focus on traditional Chinese medicine compound,traditional Chinese Medicine(TCM)has rich clinical experience in the treatment of depression,also can improve the accompanying symptoms of depression,with the advantage of less adverse reaction.Our group preliminary studies confirm the exact antidepressant effects of xiaoyaosan,and through the system research of solvent extraction and the corresponding effect validation,obtained the compound Compound Chaigui Fang with exactly antidepressant effect.On the basis of determining optimal extraction process for Compound Chaigui Fang,this paper using stable pilot plant extracts studied the comprehensive safety and efficacy.Objective:Using stable pilot plant extracts of Compound Chaigui Fang preliminarily study the safety;Selecting antidepressant effective dose of Compound Chaigui Fang by pharmacodynamic experiment;Using a variety of depression animal models comprehensively evaluate the antidepressant effect of Compound Chaigui Fang,and then on the basis of effect study the characteristics of its effectiveness;Through the metabonomics technology to validate its effectiveness and discuss its possible antidepressant mechanism.Methods:According to the Technical guidelines for the study of toxicity of single drug administration,using 40 ICR mice with half of the male and female to study the toxicity of Compound Chaigui Fang in pre-experiment,choosing the death number or weight of mice as index.It is concluded that there is none mice died in all groups under the maximum dosage and the weight growth normally.On the basis of preliminary experiment results,using 40 ICR mice with half of the male and female to study the maximum tolerance dose of Compound Chaigui Fang,choosing the mice weight and the blood biochemical of reflect the liver and kidney function as indicators.Based on previous research results and the clinical dose of xiaoyaowan(enrichment),design multiple dose group to scree the antidepressant effective dose,using the mouse tail suspension test(TST)and mice forced swimming test(FST).Using TST in mice,FST in mice,reserpine induced depression model in mice and rat chronic unpredictable mild stress(CUMS)model of depression,comprehensively evaluate the antidepressant efficacy of Compound Chaigui Fang.On the basis of CUMS model,using Morris water maze evaluate the improvement effect of Compound Chaigui Fang in cognitive dysfunction appeared on CUMS rats;using Elevated Plus Maze evaluate the improvement effect of Compound Chaigui Fang in anxiety appeared on CUMS rats;with the index of manure quantity and character evaluate the improvement effect of Compound Chaigui Fang in abnormal feces of CUMS rats.In the last day of the experiment,acquisition of rat serum detected by ~1HNMR,analyzes each rat serum metabolites with the application of metabonomics technology and the method of multivariate statistical analysis,for finding out the potential of depression disease markers and the treat markers of Compound Chaigui Fang,to evaluate the antidepressant efficacy of compound wood is belong to the party and to explore its possible mechanism.Results:1.Using maximum concentration of Compound Chaigui Fang and the mice gastric volume 40 ml/kg to ICR mice,there is none mice died in 7 days and the weight growth normally,so we could say max dosage of Compound Chaigui Fang is 140 g/kg;According to the results above,mice were fived max dosage twice a day,there is none mice died in 14 days and the weight growth normally,so we get a conclusion that the maximum tolerance dose of Compound Chaigui Fang is more than 280 g/kg.2.The results of FST and TST show that the Compound Chaigui Fang begin to work in doses for 7 g/kg,in dose of 14 g/kg and 28 g/kg markedly effective,but the xiaoyao pill(enrichment)in the equivalent dose of clinical dosage there is no significant difference compared with the blank group,so we could say xiaoyao pill under current clinical dosage cannot have antidepressant effect,need to increase the dose.3.The results of FST and TST show that the Compound Chaigui Fang could significantly reduce the dead time(P<0.01 or P<0.05);Compound Chaigui Fang could significantly improve weight loss,the reduce rate of sugar water preference and the weakenss of independent activity of CUMS model rats;Compound Chaigui Fang could significantly improve the depressive symptoms,like hypothermia and drooping eyelids,of reserpine induced model mice.All the results show that Compound Chaigui Fang has significant antidepressant effect.4.The results of pharmacodynamic characteristics experiments show that the CUMS model rats with anxiety,cognitive dysfunction and abnormalities of the stool.Each dose group Compound Chaigui Fang and venlafaxine can be significantly increased(P<0.01 or P<0.05)in the rat upright number,number of probe,open arm activity time and so on;could be significantly reduced(P<0.01 or P<0.05)the incubation period,significantly increased(P<0.01 or P<0.05)the crossing platform number and time in target quadrant activity indicators,but venlafaxine group there was no significant difference with model controls,can significantly improve the rat swimming speed;Middle and hight dose group)of Compound Chaigui Fang and venlafaxine can be reduce the number of defecation,adjust the moisture of faeces to normal levels,venlafaxine group and Compound Chaigui Fang low dose group excessively accommodate the moisture of faeces caused constipation.The bodyweight of each dose Compound Chaigui Fang were higher than that of venlafaxine group,showed that the Compound Chaigui Fang could adjust CUMS rats gastrointestinal function better.5.Based on results of ~1H-NMR metabonomics analysis,it find out that N-acetyl glycoprotein,choline and glucose significant rise(P<0.05 or P<0.01)compared with control group rats serum,and the isoleucine,trimethylamine oxide and creatine content decreased significantly(P<0.05 or P<0.01).The groups of Compound Chaigui Fang and venlafaxine could adjust the six difference metabolites close to normalConclusion:The results of toxicity experiment shows that the security of Compound Chaigui Fang is goog;The pharmacodynamics experiment indicate that Compound Chaigui Fang has obvious antidepressant effect,and could improve depression comorbid,like anxiety,cognitive dysfunction and abnormalities of the stool,and the role of play maybe through adjusying the sugar metabolism,amino acid metabolism,and the regulation activities of intestinal flora. |
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