(一)Pharmacognosic And Taxonomic Studies On Medicinal Plants Of Huperziaceae From China (二)Studies On Antidepressant Activity Of Chinese Endemic Herbal Medicine Piper Laetispicum C. DC.

AS the one of the most ancient families of vascular plants on the earth, Huperziaceae, which had been flourishing in Paleozoic, is now slowly eroding away because of angiosperm's growing up as well as the worsening of entironment. Little attention had been paid to this kind of slim and weak plant until Huperzine A, the most effective Acetylcholinesterase inhibitor among natural products, was separated by Chinese scientists from Qiancengta, a herbal drug in the South China. From then on, An accelerated overmuch exploitation began to descend the survival of Huperziaceae. In comparison with prosperous phytochemistry and pharmacology studies on Huperzine A, Few people was interested in the research on the Huperziaceae so that there are still much more vacancy in the field of basic study. In order to illuminate several pharmacognosie problems before this kind of herbal drugs are thorough destroyed, follow experiment were complished:1. More than six months were aggregately used to collect specimen and to investigate the plant resouces of Huperziaceae, and then 26 species belonging to all the five sections of two genera, Phlegmariurus (Herter) Holub and Huperzia Bern., were respectively obtained as a result. Furthermore, 2 new species and a new variety, named Huperzia hunanensis S.L.Pan et J.Wang sp.nov., H. minxiensis J. Wang et S. L. Pan sp. nov. and Huperzia liangshanica var. liaodongensis J.Wang, Y. Zhou et S.L.Pan var. nov., were successfully appreciated by two famous pteridologists.2. Having projected all species to taxonomic research, it was found that spore morphology of these species under scanning electron microscope (SEM) is not only the most steady and powerful taxonomic proof forHuperziaceae to differentiate between Phlegmariurus (Herter) Holub and Huperzia Bern., but also our consequent standpoint to support Holub J., who suggest that Huperziaceae be separated into two genera instead of a single Huperzia Bern, in 1964. By the way, Holub's theory has not been accepted by almost all the foreign pteridologists even including Holub himself for serious lack of overwhelming proof of taxonomic characters to distinguish Phlegmariurus (Herter) Holub from Huperzia Bern.. To persuade people to admit the rationality of Phlegmariurus (Herter) Holub, no uttermost technology is necessary, the spore morphology under SEM is powerful enough.A key to 21 species of Huperziaceae was then established in virtue of the spore morphology characters alone, while the other two species, Huperzia. chinensis (Christ) Ching and H. serrata (Thunb. ex Murray) Trev. f. intermedia(Nakai) Ching are repealed because they can not be taxonomically told from H. appressa (Desv.) Love et Love and H. sutchueniana (Herter) Ching, respectively. Based on the total morphologic characters of both whole plants and their spores, H. appressa (Desv.) Love et Love, which is now mergered by H. selago (L.) Bernh. ex Schrank et Mart., was able to be taken as a natural species. Spore morphology characters also supported the credibility of 2 new species and 1 new variety.3. In order to estimate the quality of those medicinal plants from Huperziaceae, a HPLC method was established to determine Huperzine A in 23 plants of Huperziaceae. The contents of Huperzine A determined were very high in Huperzia. appressa (Desv.) Love et Love and Phlegmariurus cryptomerianus (Maxim.) Ching, which was found to be 0.1710% and 0.1961% respectively, while no Huperzine A was found in H. crispata (Ching ex H.S.Kung) Ching, H. miyoshiana (Makino) Ching, H. nanchuanensis (Ching et H.S.Kung) Ching et H. S. Kung and Ph. sqarrosus (Forst.) Love et Love.If this method is modified, Huperzine B, another important alkaloid from Huperziaceae, could be identified together with Huperzine A easily.4. Concerning Chinese herbal medicine Qiancengta that was now usually used as raw materials to obtain Huperzine A, it is well accepted that its original plant is Huperzia serrata (Thunb. ex Murray) Trev.. In this paper, textual research of Qiancengta resorted to SEM and HPLC, and then we managed to clarify that the original plant of Qiancengta was H. javanica (Sw.) Chun-Yu Yang, instead of H. serrata (Thunb. ex Murray) Trev. or H. crispate (Ching ex H. S. Kung) Ching. It was also unpuzzled that H. serrata (Thunb. ex Murray) Trev. was a species whose distribution had been unconsciously enlarged during the interpenetration between Chinese botanist and Japanese scientists, in addition, the rightful taxonomic degree H. javanica (Sw.) Chun-Yu Yang should be resumed as a natural species.Depression is an increasing disease among modern society and is terribly threatening people's health and mind. Recently natural antidepressant medicines with lower toxicity and lower price than chemical medicines become more and more popular among researchers and consumers in different countries, for example, many plants belonging to Piper L., especially Piper methysticum Forster (kava-kava), have been very well accepted all over the world. China possesses more than 60 species of genus Piper L. including many endemic ones as varies of herbal medicines there. The former studies of our group had successfully drawn a conclusion that Piper laetispicum C. DC, looked as a folk herbal medicine in Hainan island of China, had distinctly more strong antidepressant activity than fluoxetine or kava-kava extract.This paper tried to probe into its antidepressant effects , active components and the pharmacological mechanism from effective fraction to pure compounds.1. We have invested the resources, distribution and folk using of Piper laetispicum C. DC. , appraising the original plant and crude medicinalmaterials ,and find that it has vivid characteristic different from the fakes.2. Contrasted with fluoxetine and extract of kava-kava, pharmacological effects of the most effective fraction (EtOAc extracts by solvent-extraction methods ) of Piper laetispicum C. DC. was further investigated through classical animal model,. Other two classical experimentations were employed to validate that the effects were not caused by their exhilarant action .3. Three compounds are isolated from the EtOAc extracts of Piper laetispicum C. DC. using silica gel column chromatography. One of them is a new compound named laetispicine, whose antidepressant effect was 5 times of fluoxetine. The other two compounds were obtained in plant for the first time. We have applied for patents of laetispicine and effective fraction of Piper laetispicum C. DC..4. In order to control the quality of crude drugs and to find the substitute of Piper laetispicum C. DC, a stable HPLC method was established to measure laetispicine in plants of Piper L. from China. The results of the contents of EtOAc extract and laetispicine show that fluctuation is common, just because series of difference in producting areas, harvest seasons, mature degree, et al. laetispicine wasn't detected in other plants of Piper L. from China.5. The antidepressant mechanism was lucubrated about both the extractof Piper laetispicum C. DC. and laetispicine, in virtue of nucleus drug method. It was illuminated by determination of the reuptake of H~3 labeled 5—HT DA and NE that The antidepressant mechanism of laetispicine and EtOAc extracts of Piper laetispicum C. DC. was a result of the total inhibitory effects on the three H~3 labeled monoamine transmitters above. Then we have applied for Chinese and international patents about these mechanism, too.