| Ilex latifolia Thunb (I. latifolia) is derived from the leaves of Folium llicis latifoliae and mainly distributed in the region of South China, which is a kind of traditional Chinese beverage with health care function.I. latifolia contains many kinds of chemical constituents, such as saponins, polyphenols, flavonoids, caffeine, triterpenoids, etc. The tea has a bitter taste, followed by cool and sweet. It has various pharmacological activities with antihypertensive, lose weight, antitumor, anti-aging, and quicken the blood vessels. In recent years, an increasing research on I. latifolia was mainly concentrated in the health protection of tea and beverage, but the study of functional components in I/. latifolia remained to be further in-depth discussion, which was in order to promote the development and utilization of tea resource. The bioactive compounds of I. latifolia were purified and identified in terms of modern chromatographic and spectroscopic techniques. The underlying mechanisms of triterpenoid-induced inhibitions of human breast carcinoma cells (MCF-7) proliferation were illustrated. The adsorption kinetics and thermodynamics of adsorption processes by macroporous resin for I. latifolia phenolics were also studied. The anti-inflammatory effect of I. latifolia phenolics were investigated. The extraction, purification, physico-chemical properties, structural analysis and evaluation of immunobiologic activity of I. latifolia polysaccharides were illustrated.(1) Use high performance liquid chromatography (HPLC) to establish the fingerprint for 10 batches of I. latifolia under the optimization extraction and chromatography methods. Evaluate the similarity of 10 batches samples by "traditional Chinese medicine (TCM) fingerprint similarity evaluation system" software in order to establish the common model of fingerprint and obtain the sililarity between each sample and common model. The results showed that the relative retention time of common peaks among the 10 batches of I. latifolia were consistent, and RSD was less than 1%, the similarity was greater than 0.9. It conformed to the requirements of the TCM fingerprint technology. The common model could be better to reflect the distribution of chemical composition in I. latifolia.(2) The active ingredients was obtained from the I. latifolia, which was refluxed with ethanol in a water bath. Then the total extract was portioned with petroleum ether, ethyl acetate, n-butanol and water, respectively, in order to get four different polar components. Results showed that ethyl acetate fration not only had excellent antioxidant activity and good resistance to complement activity, but also had significant inhibition of MCF-7 proliferation activity, which indicated that the main active ingredients were concentrated in the ethyl acetate fration after solvents grading extraction.13 compounds were purified from the ethyl acetate fration by various column chromatography and their structures were identified in terms of nuclear magnetic resonance spectrum (NMR) and mass spectrum (MS) as ethyl caffeate, β-sitosterol, ursolic acid, chlorogenic acid, β-daucosterol, isochlorogenic acid A, Isochlorogenic acid B, Isochlorogenic acid C, myricitrin, pomdid acid (PA), EGCG,3,4-di-O-caffeoylquinic acid methyl eater,3,5-di-O-caffeoylquinic acid methyl eater. It was the first time that 3,4-di-O-caffeoylquinic acid methyl eater and 3,5-di-O-caffeoylquinic acid methyl eater were found in I. latifolia.(3) Among all compounds isolated from ethyl acetate fraction, PA showed the better inhibitory effect of two kinds of tumor cells and two kinds of normal cells determined by MTT experiments, especially on MCF-7 cells, which showed strong cells selectivity. PA inhibited MCF-7 cell proliferation by inducing apoptosis and G2/M phase arrest through the flow cytometry analysis. The underlying mechanisms of PA-induced inhibitions of MCF-7 proliferation were studied by means of intracellular ROS level detection, JC-1 staining detection, real-time fluorescent quantitative PCR and western blot. Results indicated that PA showed antiproliferative activity and induce the apoptosis of MCF-7 cells by significantly increasing the content of intracellular reactive oxygen specied (ROS) level to break the balance between oxygen metabolism in cells, reducing cell mitochondrial transmembrane potential to cause mitochondrial dysfunction, increasing the Bcl-2, CDK4, Cyclin D1 mRNA expression and reducing the p53 mRNA expression.(4) The best extraction technology of I. latifolia polyphenols were studied and optimized. The antioxidant capacity (remove DPPH free radicals and ABTS+ free radicals, total antioxidant capacity, total reducing power) of I. latifolia polyphenols was comprehensive evaluated and the experimental results showed that it has excellent antioxidant activity. Preliminary study was found that the enzyme activity of I. latifolia polyphenols possessed good inhibitory effect through a-glycosidase enzymes and β-amylase associated with digestion. The adsorption kinetics and thermodynamics of adsorption processes by AB-8 and HP-20 resins for I. latifolia polyphenols were studied. According to study the relationship between adsorption amount and temperature of the two types of resin, it found that the best temperature of AB-8 and HP-20 for I. latifolia polyphenols adsorption were 35 ℃ and 30 ℃, respectively.(5) The anti-inflammatory effect and underlying mechanisms of I. latifolia polyphenols was studied, and the experimental results showed that I. latifolia polyphenols exerted anti- inflammatory effect may be through inhibiting the secretion of NO, TNF-a, IL-6, IL-1β by LPS-induced macrophage, inhibition of intracellular TNF-a, IL-6, IL-1β, iNOS, cox-2 mRNA expression, inhibition of MAPKs signaling pathways and the NF-κB signaling pathways to modulate inflammatory factor.(6) The best extraction technology of I. latifolia polysaccharide were studied and optimized.I. latifolia polysaccharides were purified and concentrated by DEAE-52 cellulose ion exchange column chromatography in order to obtain three kinds of polysaccharides. The physical and chemical properties and structure analysis of the purified polysaccharides was investigated. In addition, the immunocompetence experimental results showed that I. latifolia polysaccharide exerted immunological activity may be through up-regulation of intracellular TNF-a, IL-6, IL-1β, iNOS mRNA expression, up-regulation of MAPKs signaling pathways and inhibition of the NF-κB signaling pathways, thus to promote the secretion of NO, TNF-a, IL-6 by LPS-induced macrophage in order to activate macrophage participate in complex immune activity, thus played the role of immune activity. |
PDF Full Text Request