| Laporte bulbifera(Sieb.et Zucc.)Wedd(also known as Honghema in Chinese)is a conventional herbal medicine native to Miao and other minorities in Guizhou Province of China.The fresh or dried plant of Laporte bulbifera was one of the most widely used Traditional Chinese Medicine(TCM)for removing dampness and promoting water metabolism for thousand years.Clinical studies showed that this herb could lower plasma cholesterol levels,protect the hepatic system,and increase the urinary excretion of sodium,chloride and urea.The chemical components of Laporte bulbifera mainly include catechins,flavonoids,tannins,coumarins,volatile oils,sugars and their glycosides,amino acids and peptides,vitamins and various trace elements.At present,most of recent research were the studies on the determination of the content of the above components and fewer compounds have been isolated and identified.The current research on the pharmacological effects of Laporte bulbifera focuses on its analgesic,anti-inflammatory and immunosuppressive effect.In this research,each male KM mice was fed high-fat diet for two weeks to establish mouse hyperlipidemia model.Laporte bulbifera was extracted,separated and concentrated to obtain four fractions of water fraction,30% ethanol fraction,70% ethanol fraction and 95% ethanol fraction.Each mice in treatment was intragastrically administered with a dose of 40 mg·g-1,respectively.After 4 weeks administration,the body weight of mice before and after experiment and the serum biochemical indexes were recorded.The liver segments from each mouse were quickly removed and prepared for oil red O histological analysis.The body weight of mice before and after the experiment and its liver index of each mice were recorded.The serum total cholesterol(TC),low density lipoprotein cholesterol(LDL-C),malondialdehyde(MDA),superoxide dismutase(SOD)in mice serum from every group were also recorded.The results of experiment showed that the 30% ethanol and70% ethanol fractions of Laporte bulbifera significantly reduced the body weight of hyperlipidemia mice and he serum levels of TC,LDL-C,and MDA in hyperlipidemia mice.The SOD level in the mice increased in the 30% ethanol and 70% ethanol fractions group.Meanwhile,the gain body weight of the mice has the same trend.The microphotographs of oil Red-O staining of hepatic tissues of mice shows the section of hepatic tissue in normal mice exhibiting the normal hepatocytes with sinusoidal cards around the central vein and portal tracts.However,the deformation of the hepatocytes around the central vein,congestion of sinusoids around central vein regions,microvesicular and macrovesicular changes were observed in hyperlipidemic mice.As depicted in 30% ethanol and 70%ethanol fractions group,the section of hepatic tissues of hyperlipidemic mice presented the almost normal hepatocellular arrangement around the central vein with abridged fatty changes.The results indicated liver tissues of hyperlipidemic mice can be protected and repaired by intervention with administration of Laporte bulbifera.The results showed that the 30% ethanol and 70%ethanol fractions of Laporte bulbifera could regulate the lipidmetabolism in experimental hyperlipidemic mice and reduce their blood lipid levels.The 70% ethanol fraction was analyzed by UPLC-QTOF-MS with a 10?L injection.UPLC separation was achieved on a reversed-phase Welch Ultimate UPLC C18 column(100 mm×2.1 mm,1.8?m).The gradient elution of A was water-formic acid(99.9: 0.1,v/v)and B was acetonitrile.The detection wavelength was set at 190-800 nm.The most intensive of 8 ions from each full MS scan were selected as precursor ions for MS/MS fragmentation in the information dependent acquisition(IDA)experiments.Dynamic background subtraction was used to match the IDA criteria.Positive detection mode was used to obtain the high resolution molecular information.The and MS/MS CID collision product ion fragments were selected to identify the main chromatographic peaks.The main components of 41 kinds of Laporte bulbifera column chromatography anti-hyperlipidemia effective fractions were identified by LC-MS/MS method,including 11 Coumarins,6 oligopeptides and 24 fatty acids.To investigate the mechanism of hypolipidemic effect of Laporte bulbifera,the serum of the mice administered by 70% ethanol fraction was analyzed by UPLC-QTOF-MS/MS.Principal component analysis(PCA)and partial least squares(PLS-DA)were used to explore the differential changes in metabolites in mice serum.Four potential hypolipidemic biomarkers were found.The KEGG database was used to screen its associated metabolic pathways and six related pathways were found.The experimental results show that the hypolipidemic effect of Laporte bulbifera may come from the regulation of lipids,carbohydrate,energy and amino acidmetabolism in vivo to regulate blood lipids.In this research,the water extract of Laporte bulbifera was separated and purified by D101 macroporous resin column chromatography.The fractions were administered to experimental hyperlipidemia model mice to investigate the active anti-hyperlipidemia fraction of Laporte bulbifera.The anti-hyperlipidemia components were determined by UPLC-QTOF-MS/MS.The active ingredient group was identified or characterized and the mechanism of the hypolipidemic effect was explored through metabolomics techniques.It laid the foundation for further research on the effective components of anti-hyperlipidemia components in Laporte bulbifera,and also provided a scientific basis for the research and development of traditional Chinese medicine for anti-hyperlipidemia. |
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