| Depression is a common mood disorder with obvious and lasting depressing state of mind as its main clinical features, and more severe patients have suicidal thoughts and behavior. Depression, cancer and AIDS are considered to be the three major diseases in the new century. With the increasing pressure of the work and living, patients with depression have increased year by year, which has become a global social and economic issue that must be resolved. Its pathogenesis is extremely complex and there is no precise explanation in domestic and abroad, the hypothesis based on the monoamine has promoted the progress of the depression.Chaihu shugan powder(CSGS) is the representative prescription of soothing liver and regulating vital energy, consisting of Chinese Thorowax Root, Tangerine Peel, Szechwan Lovage Rhizome, Nutgrass Galingale Rhizome, Submature Bitter Orange, White Peony Root and Liquorice Root. It has got good clinical effect in the treatment of depression while the mechanism still remains uncertain. Traditional method for medication is decoction or powder. Our group optimized the process for CSGS extraction in preliminary work. Compared with aqueous extraction, the active ingredients of CSGS by alcohol was significantly higher and the material composition also changed. In this study, the ethanol extraction was prepared and its therapeutic effect on depression and the possible mechanism by the behavior of the mice model of depression, the content of monoamine neurotransmitters and their metabolites as well as antioxidant experiments in vitro and in vivo were studied. 84Kunming mice were divided into7groups at radom:normal group, model group, Water formulation group(WF group), the same content of saikosaponin group(1.6g·kg-1, SA group), alcohol extract group(AF group)4.1,8.2,16.4g·kg-1dose group. The experiments below were investigated.1The effect on behavioral indices of depression miceCopy the depressive model of mice by CUMS combined with CRS to evaluate the behavioral experiments such as the body weight growth rate, sucrose preference test, open field test, emotional behavior test, tail suspension test and force swimming test.The results showed that CUMS and CRS could induce the mice depression. Compared with the normal group, the body weight growth rate in model mice was more slowe(P<0.01); in sucrose preference test, the sucrose preference rate in model group reduced obviously; in open field test, the acrossing-movement times and rearing-movement times of model mice were lower(P<0.01, P<0.05); in emotional behavior test, the emotional behavior score of model mice was lower(P<0.05); in tail suspension test, the non-movement time of model mice was longer(P<0.01); in force swimming test, the immobility time of model mice increased(P<0.01). Compared with the model group, water extract group, CA group and alcohol extract groups (4.1,8.2,16.4g-kg-’) could improve the symptoms of depressive mice with a dose-related charactr(P<0.01, P<0.05). With the same amount of crude drug of8.2g·kg-1, the mice in alcohol extract group had more body weight in the28th day compared with water extract group(P<0.05). In open field test, the acrossing-movement times in alcohol extract group were significantly higher (P<0.05) and the immobility time was lower in force swimming test(P<0.05).2The effect on the content of monoamines neurotransmitter and their metabolites in mice’s brian tissueThe content of monoamine neurotransmitters and their metabolites in brain were detected by high performance liquid chromatography-electrochemical method. The Column was Hypersil BDS C18(4.6mm×250mm,5μm) and the column tempture was kept at40℃. The mobile phase, consisting of methanol-water phase(10:90), was at a flow-rate of0.8ml·min-1. The voltage of electrode was600mV. The inject volum was10μl.The results showed that the contents of norepinephrine(NE), dopamine(DA) and serotonin(5-HT) in mice’s brian tissue decreased obviously in model group(P<0.05, P<0.01); the metabolites3,4-dihydroxyphenylacetic acid and serotonin hydrochloride increased (P<0.05); The DOPAC/DA and5-HIAA/5-HT were significantly higher (P <0.01). Compared with the model group, the contents of monoamines neurotransmitter and their metabolites in every drug group were call-back with varying degrees; in8.2and16.4g·kg-1group of alcohol extract, DA increased(P<0.05); the5-HT increased in16.4g·kg-1alcohol extract group (P<0.01); DOPAC decreased in every drug group(P<0.01, P<0.05); there is significant difference for5-HIAA between water extraction and8.2g·kg-1alcohol extract group(P<0.05). DOPAC/DA and5-HIAA/5-HT group in every drug group decreased obviously except SA(P<0.01, P<0.05).3The effect on SOD and CAT oxidase activity in mice’s liver tissueThe oxidase activity in liver tissue and antioxidant capacity of alcohol extracts in vitro were measured by using ultraviolet-visible spectrophotometry. The SOD was detected by Pyrogallol. The CAT was done by ammonium molybdate.The results showed that SOD and CAT oxidase activity in mice’s liver tissue decreased obviously in model group(P<0.01). Compared with model group, the SOD oxidase activity increased significantly in the8.2,16.4g·kg-1group of alcohol extract(P<0.01); the CAT oxidase activity increased in every drug group(P<0.05, P<0.01). Compared with water extract group, the two kinds of oxidase activity of8.2g·kg-1group in alcohol extract both increased obviously(P<0.05, P<0.01). 4The antioxidant activities of Chaihu shugan powder of alcohol extract in vitroThe anti-oxidant activity in vitro tissue was measured by using ultraviolet-visible spectrophotometry. We detected DPPH,·OH, O2-· and Vc was the positive control ones.The results showed that the IC50of CSGS in alcohol extract was0.75μg·ml-1(DPPH),35.01μg·ml-1(·OH),52.17mg·ml-1(O2-·) respectively; the IC50of Vc was5.79μg·ml-1(DPPH),0.108mg·ml-1(·OH) and0.152mg·ml-1(O2-·). The IC50of CSGS alcohol extract was even lower than Vc.In summary, we copied the mice depression model successfully by CUMS combined with CRS, and then it was proved by behavioral experiment. Meanwhile, it also indicated that the CSGS alcohol extract has the effect of anti-depression. CSGS can ameliorate the metabolic disorders of DA and5-HT, but it has not an obvious effect on NE. CSGS has better anti-oxidation activity in vivo and in vitro, which may be related to its anti-depression mechanism. |
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