| Ficus microcarpa L.f. are from Moraceae Ficus drying the leaves, the water extract ofdry extract as the main ingredient Keteling capsules were contained in the ministerialstandards"Ministry of Health standards of the PRC"(Volume XIV medicine prescriptionpreparations). Which could crue the asthma and chronic bronchitis. Besides, the clinicalcurative effect are exact. Considering the multicomponent of traditional Chinesemedicine(TCM) and the overall function, the fingerprint method is applied to controlChinese medicine extract as much as possible. But the curative effect of TCM is the overallperformance, which stands for functional components. Besides, the overall effect is theresult of multiple ingredients targets synergy, neither a single component and simpleaddition of ingredients. In this paper, we study the quality control model based on thechemical composition and efficacy research, expounding the correlation between"composition and efficacy" and building one or several active ingredients of TCM inqualitative and quantitative analysis method, which can specify the quality of the medicinalingredient so as to establish a scientific and reasonable quality control methods, in line withthe requirements of modern medicine. The main contents:1. The RAW264.7macrophage line was induced by lipopolysaccharide(LPS) to set up theinflammatory model. MTT assay was performed to determine the viability of the water extract of Ficus microcarpaL.f and the polar solvent extract of different parts on RAW264.7;Griess reaction was used for assaying the concentration of Nitric Oxide(NO); ELISA wasperformed to establish the effects on Tumor Necrosis Factor-?(TNF-?) production. ResultsNO and TNF-? contents in medium of macrophages were significantly increased after thestimulation by LPS. Among of them, the water extract of Ficus microcarpa L.f and thepolar solvent extract of different parts could remarkable ameliorate the inflammationfactors release under the different levels, which the water extract of Ficus microcarpaL.f (atthree concentration groups), the acetate extract(low-concentration group andmedian-concentration group) and water layer(low-concentration group) were exertedsignificant effects. Ficus microcarpa L.f played a anti-inflammatory role on inhibiting therelease NO and TNF-?. The release of inflammatory mediators were compared with eachother comprehensively. Therefore, the ethyl acetate layer and and water layer could be usedas an anti-inflammatory effective part, which could provide a reference for extraction andseparation of Ficus microcarpa L.f.2. Appling with the serum chemistry method, the components into blood of FicusmicrocarpaL.f were analysed, laying the foundation for studies on potential basis materials.The fingerprints in vitro and HPLC fingerprint analysis after gavage containing serum wereestablished; through contrast fingerprints in vitro, the part into the blood was confirmed,which determined the site for the next step to analyze the chemical composition and theseparation of blood components into the narrow scope of analysis. The results showed thatthe drug-containing serum showed seven ingredients into the blood, blood components intoroughly determine the polarity of the main parts of the ethyl acetate layer. The part couldsupply a specified direction for the latter part of effective substance.3.The extraction and seperation were conducted on chemical constituents ofanti-inflammatory effective fraction of Ficus microcarpa L.f. Structural determination wasperformed on the separated chemical constituents. we had identified and evaluated theactivity ingredients from the gained chemical constituents. To separate and purify theeffective fraction, silica gel column Chromatography was adopted. The structures ofconstituents were identified by IR,1HNMR,13CNMR. The RAW264.7macrophage line was induced by lipopolysaccharide(LPS) to set up the inflammatory model. MTT assay wasperformed to determine the toxicity of constituents on RAW264.7; Griess reaction was usedfor assaying the concentration of Nitric Oxide(NO); ELISA was performed to establish theeffects on Tumor Necrosis Factor-?(TNF-?) production. Ten compounds were prepared,Compared with model group, the situation of inhibition of NO and TNF-?, which TheRAW264.7macrophage line was induced by lipopolysaccharide(LPS). Ten monomercompounds were isolated from the effective parts of the Ficus microcarpa and identified. Inaddition, the experiment confirmed the anti-inflammatory pharmacological actions could beimpacted by multiple components and indexs. The result further reveals the material basisfor anti-inflammatory efficacy and provides a reliable theoretical guidance and credibledatas.4. To study the HPLC fingerprint chromatogram for the leaves of Ficus microcarpa collectedin different harvest periods could provide the basis for the identification of FicusmicrocarpaL.f and the optimization of the best harvest periods. The fingerprint chromatogramcould be divided into high polarity part and low polarity part by the peak No.29, which wasidentified as reference peak of sec-butyl-4-hydroxybenzoate. The chromatograms of12batches of traditional Chinese medicinal materials in different harvesrt periods weredetermined by HPLC, which existed a total of about55chromatographic peaks. Among themthere were44chromatographic peaks in common. The peaks were named as characteristicfingerprint peaks. At the same time, we conducted a correlation analysis and regressionanalysis between44common peaks in12batches of herbs and efficacy data of NO andTNF-? inhibition rate using statistical software. Finally we found the composition group,which played a pretty efficacy in vitro, so as to establish the quality standards for the waterextraction of Ficus microcarpa L.f.5. In uniform chromatographic conditions, we can get the chromatographic fingerprint ofeffective parts from the12batches of Ficus microcarpa L.f. in vitro and the12batches ofchromatographic data, we conduct the correlation analysis and regression analysis betweenthe area value from effective parts in the chromatographic fingerprint chromatography invitro and the efficacy data of NO and TNF-? using SPSS statistical software. In the end, we can determine the composition of the anti-inflammatory effect of generation and in vitrowhich are closely related. Spectral efficiency by correlation analysis using statisticalsoftware, combined with the extraction of chemical constituents isolated, so the threegroups to determine the content medicinal ingredients, in order to establish a controlmethod on the basis of efficacy, improve the quality of Keteling Capsules. |
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