Study On Screening Of Anti-inflammatory Active Site And Components Of Lepisorus Ussuriensis(Regel Et Maack) Ching

Herb Lepisorus ussuriensis, alias is stone tea or tree tea. It is the herb of Lepisorus ussuriensis (Genus:Lepisorus; Family:Polypodiaceae), located in the northeastern and Hebei, Shanxi, Shandong, Sichuan and other places in China. According to records, there are has the effect of detoxification, diuretic, cough, bleeding, swelling in herb Lepisorus ussuriensis. It is widely used in folk.By investigation, there are abundant resources of Lepisorus ussuriensis in China. It has numerous bioactive compounds, medicinal and considerable economic value. It is a kind of natural medicine not to mining fully, so it is a medicinal plants with further research and development. We did the following experiments:First of all, among Lepisorus there are several species, the rough classified. Therefore, in order to ensure the accuracy crude drugs, has carried on the pharmacognostic identification. Pharmaeognostic researches aimed at the identification of original, macroscopical, and physicochemical identification, established as the the following identification basis:①The slender rhizomes is cross growth with dense black or dark brown hyaline scales; blade is usually coriaceous. The leaf is0.5～1cm width in the central,1～5cm length of the petiole, the circular sporangium.②Crimping and slender drying herbs is fragile, the main vein is raised on upper and lower epidermis, minor veins is netted;③Epidermis cells of rhizome are arranged in a row and several layers of thick cells beneath the epidermis cell, dictyostele, there are1vascular bundle with the xylem of "T" shape in the middle of the petiole. There are are mainly polar cell type and polar cell type with the same circum on the stoma types.④HPLC method was employed to determine the content of chlorogenic acid and ecdysterone and lutein.The content of chlorogenic acid was2.635mg/g, ecdysterone was2.509mg/g, lutein was110.0μg/g. The three methods is accurate, sensitive and easy for determination chlorogenic acid, ecdysterone and lutein in Herb Lepisorus ussuriensis.Secondly, screening the anti-inflammatory active site of Herb Lepisorus ussuriensis:. Eevaluate the anti-inflammatory effect to inhibit mouse ear swelling by xylene induced on the extract of the different doses of water extract and70%ethanol extract, the result showed the best anti-inflammatory effect that was the high dose group of70%ethanol extract. Then, screening the best anti-inflammatory activity site, using different solvent extraction and gradient extraction method to70%ethanol extracton of Herb Lepisorus ussuriensis, the results showed, the n-BuOH extract of ethanol extract was the highest of its inhibition rate on mouse ear swelling, and compared with model group, P<0.01, with aspirin control group, P<0.05. It showed that n-butanol extract of ethanol extract is the strongest anti-inflammatory activity site. Thirdly, in order to determine the marker components of anti-inflammatory activity site, to establish the detection method of quantitative, to determine of the chlorogenic acid and ecdysterone in the different extracts by HPLC method:The results showed that the content of chlorogenic acid and ecdysterone was the highest in n-butanol fraction of ethanol extract. But the swelling inhibition rate increased with increasing of chlorogenic acid in the extract, chlorogenic acid explain is maker component of the anti-inflammatory.Finally, optimize the extraction process of chlorogenic acid in Herb Lepisorus ussuriensis. Using response surface method optimized the extraction process of hot water extraction and ethanol extraction method. The results showed that ethanol extraction, extraction time41min,75%ethanol and the ratio of solid to liquid1:68is the best extraction process. The extraction rate of chlorogenic acid can reach8.260mg/g.By this research, provide the identification basis and scientific basis for its estimation of quality of Lepisorus ussuriensi. Screened out of the best active fraction on anti-inflammation, provide a new medicine source plant of chlorogenic acid for the extraction. It is established the optimum extraction process of chlorogenic acid in Lepisorus ussuriensis.