The Study Of Establishing The Fingerprint Chromatogram And Revealing The Anti-depression Effect And Its Mechanism Of Milletia Speciosa Champ

OBJECTIVE:The study aims to optimize and establish the fingerprint chromatograms of Milletia speciosa Champ by HPLC,and then evaluate the similarity of Milletia speciosa Champ samples from 13 different habitats.In addition,the study is focus on analyzing the influence on the chemical composition of Milletia speciosa Champ by different climate factors.Furthermore,the research aims to investigate the anti-depression effect and its mechanism of Milletia speciosa Champ,in order to provide experimental basis for the quality control and practical application of Milletia speciosa Champ.METHODS:1.The fingerprint chromatogram of the Milletia speciosa Champ samples were optimized and established by HPLC-PDA after investigating the optimization of chromatographic conditions and the extraction technology,and evaluated the reliability of that.2.The similarity of Milletia speciosa Champ samples from 13 different regions was evaluated by application of"Chinese medicine chromatographic fingerprint similarity evaluation system"（2004 a）.3.The change of the chemical composition of NDL was studied by partial least squares discriminant analysis,which was affected by different climate including the annual average temperature,precipitation and sunshine duration.4.Preliminary study on the anti-depressant effect of Milletia speciosa Champ by tail suspension test（TST）,forced swimming test（FST）and the open-field test（OPT）.5.The depression model of mouse was established by chronic unpredictable mild stress（CUMS）model,and the anti-depressant effect of NDL was evaluated using sugar preference test（SPT）and the open-field test（OPT）.6.To explain the anti-depression mechanism of NDL,ELISA was used to assess the influence on monoamine neurotransmitter（including 5-HT,DA and NA）and BDNF in brain of CUMS model mice after treated with Milletia speciosa Champ.Results:1.HPLC fingerprint method is established that was used to analyze the NDL.2.Through an analysis of 13 batches of Milletia speciosa Champ samples,a total of 15 common peak is to find out in the HPLC fingerprint,however,there are some NDL sample low similarity with other sample.3.Because including the annual average temperature,precipitation and sunshine time these climate factor is different,Milletia speciosa Champ chemical composition profile showing a trend of separation.4.Based on the 15 common peak in HPLC fingerprint were analyzed,and the 9,11,12 peaks are greatly influenced by temperature;1,5,11,13 peaks are greatly influenced by amount of precipitation;1,3,5,11,and 13 peaks easily affected by the sunshine duration.5.After CUMS stimulation,sugar preference text（SPT）and the open-field text（OPT）score in mice was reduced significantly（P<0.01）,and various doses of NDL(5～20 g·kg^-1)can improve the SPT（P<0.05）,and M,H-NDL can improve the autonomic activities score of mice in OPT（P<0.05）.6.After CUMS stimulation,monoamine neurotransmitter（including 5-HT,DA and NA）and BDNF in brain of CUMS model mice were decreased significantly（P<0.05）,H-NDL can reversal of 5-HT decline significantly（P<0.01）,M,H-NDL can improve the content of NA（P<0.05）,and than different doses of NDL can improve the content of BDNF significantly（P<0.01）,and the curative effect is superior to the FLU（P<0.05）.CONCLUSION:1.In this study,the fingerprint chromatograms of Milletia speciosa Champ by HPLC was established successfully,which was simple operation and easy to be used for quality control of NDL.2.During 15 common peaks were defined in the fingerprint chromatograms of NDL by HPLC,a total of six common peaks were susceptible to influence by different climatic factors.3.Milletia speciosa Champ was found significant antidepressant effect on behavioral despair animal models and CUMS model,the mechanism of that may be related to increase the level of 5-HT,NA and BDNF in mice.