RESUMEN
OBJECTIVE To extract and isolate the four chemical components of Yao medicine Ventilago leiocarpa， and to conduct identification and content determination for them. METHODS The chemical components of V. leiocarpa were separated and purified by solvent extraction， extraction， silica gel column chromatography and preparative liquid chromatography； then the chemical structures of four isolated compounds were identified based on their spectral data. The contents of four components were determined by high performance liquid chromatography（HPLC）-quantitative analysis of multi-components by single-marker （QAMS） method， with the following chromatographic conditions： chromatographic column was Echway GowonTM C18 （250 mm× 4.6 mm， 5 μm）. The mobile phase was acetonitrile-0.1% phosphoric acid for gradient elution； the detection wavelength was 269 nm， and the column temperature was 25 ℃ . Using emodin as internal reference， the relative correction factors （fi/s） between emodin and the other 3 components were established and used to calculate the content. At the same time， the content of each component was calculated with the external standard method （ESM）， and the differences between these two methods were compared. RESULTS Four compounds were isolated from V. leiocarpa， and they were identified as emodin， frangulin A， pleuropyrone A， emodin-8-O-β-D-glucoside. The result of HPLC-QAMS showed that the fi/s of pleuropyrone A， emodin-8-O-β-D- glucoside and frangulin A were 1.147 2， 0.874 7 and 0.644 4， respectively. The content of these four components was measured as a good linearity （r≥0.999 6）； relative standard deviation （RSD） of precision， stability and reproducibility tests were all lower than 2.00%， and average recoveries were E-mail：dearhuangjianyou@126.com 99.41%-100.46%（RSD≤2.05%）. There was no significant difference between QAMS method and ESM （RSD＜3.00%）. CONCLUSIONS Emodin， frangulin A， pleuropyrone A and emodin- 8-O-β-D-glucoside are isolated from V. leiocarpa； among them， the last three components are all isolated from for the first time. The established HPLC-QAMS method is accurate and reliable for the determination of 4 components in V. leiocarpa， and can used for quality control of V. leiocarpa.
RESUMEN
OBJECTIVE：To establi sh the quality standard of Yao medicine Pileostegia tomentella，and to provide reference for its quality control. METHODS ：Totally 10 batches of P. tomentella were collected from Guangxi area. The properties of P. tomentella were observed and microscopic identification was conducted for the transverse section of its stems ，roots and powder. TLC method was established by using 7-hydroxycoumarin as reference. The contents of water ，total ash ，acid-insoluble ash and alcohol-soluble extract in P. tomentella were determined. The contents of skimmin ，7-hydroxycoumarin and 7-hydroxy-8- methoxycoumarin in P. tomentella were determined by HPLC. RESULTS ：The root and stem surface of P. tomentella possessed obvious wrinkles ，hard texture ，slight smell and bitter taste ，and there were secretory cells and needle crystal cells in the cross section. The medicinal powder contained calcium oxalate needle crystal ，calcium oxalate square crystal ，sclerotic cell and starch granules. The results of TLC showed that the spots of the same color were found in the corresponding positions of chromatograms of test samples and 7-hydroxycoumarin control. In the 10 batch of P. tomentella ，the contents of water were 9.1％-11.8％；those of total ash were 3.6％-6.7％；those of acid-insoluble ash were 0.10％-0.44％；those of alcohol-soluble extract were 8.9％-14.0％.The linear range of skimmin ，7-hydroxycoumarin and 7-hydroxy-8-methoxycoumarin were 6.088-182.640，1.568-47.040 and 1.096- 32.880 μg/mL（all r not less than 0.999 7）. The average recoveries were 99.47％，101.07％ and 99.89％（RSD were all less than 2％）. RSDs of precision ，stability（24 h），reproducibility and durability tests were all lower than 2％ . The contents of 3 components such as skimmin were 1.337-9.534，0.348-2.236， 0.083-1.088 mg/g，respectively. CONCLUSIONS ：It is preli- 201705） minarily proposed that the content of water in P. tomentella is not more than 12.0％ ；that of total ash is not more than 7.0％；that of alcohol-soluble extract is not less than 8.0％； E-mail：luguoshou@foxmail.com that of ski mmin is not less than 1.30 mg/g；that of 7-hydroxy- coumarin is not less than 0.30 mg/g；that of 7-hydroxy-8-methoxycoumarin is not less than 0.06 mg/g.
RESUMEN
OBJECTIVE：To optimize ultrasonic-assisted ethanol-（NH4）2SO4 aqueous two-phase extraction technology of citru- sinol from Desmodium caudatum . METHODS ：Using the content of citrusinol as indexes ，with ethanol volume fraction ， solid-liquid ratio ，（NH4）2SO4 addition amount ，ultrasonic time and ultrasonic temperature as factors ，based on the single factor tests，Box-Behnken design-response surface methodology was used to optimize the extraction technology of citrusinol. RESULTS ： The optimized extraction technology of citrusinol included that ethanol volume fraction was 95.35％，the solid-liquid ratio was 1∶50.35 （g/mL），（NH4）2SO4 addition amount was 4.49 g，ultrasonic time was 48.7 min，ultrasonic temperature was 57.6 ℃. In 3 times of validation tests ，the extraction rates of citrusinol were 0.637 8，0.638 4，0.625 4 mg/g，respectively，which was close to predicted value（0.630 5 mg/g）. CONCLUSIONS ：The optimized ultrasonic-assisted ethanol- （NH4）2SO4 aqueous two-phase extraction technology is stable and feasible ，and can be used for the extraction of citrusinol from D. caudatum .
RESUMEN
OBJECTIVE：To study the effects of citrusinol on proliferation ，migration and the expression of skeleton-related proteins of human hepatocellular cells HepG 2，and to investigate its interaction mode with skeleton-related proteins. METHODS ： CCK-8 assay was used to detect the effects of different concentrations （12.5，25，50，100，200 μmol/L）of citrusinol on the proliferation of HepG 2 cells for 24 h. HepG 2 cells were divided into negative control group ，citrusinol low-concentration and high-concentration groups （50，100 μmol/L citrusinol）. After treated for 24 h，the migration of HepG 2 cells was detected by cell scratch test ；cell migration rate was calculated. mRNA and protein expression of F-actin ，β-tubulin and Ezrin in HepG 2 cells were determined by RT-PCR and Western blotting assay. Molecular docking software Schrodinger 2015 was used to analyze the interaction mode between citrusinol and above 3 kinds of proteins. RESULTS ：Citrusinol showed significant inhibition effect on the proliferation of HepG 2 cells （P＜0.05 or P＜0.01），in dose-dependent trend. Compared with negative control group ，cell migration， mRNA and protein expression levels of F-actin ， β-tubulin， Ezrin were decreased significantly in citrusinol low-concentration and high-concentration groups （P＜0.05 or P＜0.01）. Molecular docking results showed that the citrusinol could form hydrogen bond and hydrophobic bond with the above 3 skeleton-related proteins. CONCLUSIONS ：Citrusinol can inhibit the proliferation and migration of HepG 2 cells，the mechanism may be associated with the down-regulation of mRNA and protein expression of F-actin ，β-tubulin and Ezrin. The mode of its interaction with skeleton-related proteins may be the formation of hydrogen bond or hydrophobic bond.
RESUMEN
Aim To investigate the antidepressive-like effect of ethyl alcohol extract of Polyrhachis vicina Rog-er(EAPR),and its mechanism.Methods EAPR was prepared by ethanol extraction.Its anti-depressive effect was investigated by tail suspension test (TST) and forced swimming test (FST).Furthermore,repeated doses of reserpine was used for preparing the depres-sive rats.Results EAPR has definitely anti-depres-sive effect,as evidenced by the decreased immobility time in FST and TST at the doses of 8 and 4 g·kg -1 (P <0.05).In the repeated reserpine evoked depres-sive rats,EAPR antagonized the symptoms induced by monoamines depletion and attenuated the anhedonia, as manifested by reversed hypothermia,akinesia and sucrose consumption at the doses of 8 and 2 g·kg -1 (P <0.05,P <0.01).Neuro-chemical studies showed that AFPR significantly increased the concentration of monoamines,including 5-hydroxytryptamine (5-HT) and noradrenaline(NA)at the dose of 8 g·kg -1 (P <0.05),and had no effect on normal rats .Furthermore, EAPR increased the activity of superoxide dismutase (SOD)in serum,hippocampus and cerebral cortex at the dose of 8 g·kg -1 (P <0.05).Conclusion EA-PR possesses the definite antidep ressive properties, connected with the regulation of neurotransmitter me-tabolism and the nerve cells antioxidant effect.
RESUMEN
Objective To investigate effect of sodium cyanide(NaCN)at different concentrations on the mitochondrial respiratory oxygen consumption,the mitochondrial membrane potential(MMP)and activity of the complex Ⅳ of mitochondrial breath train in in vitro liver mitochondria from the rats exposed to simulated high altitude hypoxia,and to explore the characteristics of energy metabolism in the mitochondria from the rats subjected to cyanide poisoning during acute hypoxia exposure.Methods Adult Sprague-Dawley(SD)rats were set randomly into control and acute hypoxia groups(n=8 in each group).The acute hypoxic rats were exposed to simulate 5 000 m high altitude in a hypobaric chamber 23 h/d for 3 d.Rats in the control group were bred in the normoxia condition at the same time.The liver mitochondria were isolated by centrifugation.Mitochondrial oxidative respiratory consumption and activity of the complex Ⅳ was measured by Clark electrode after the treatment of NaCN at 0,0.01,0.1 and 0.25 mmol/L respectively,so as to calculate mitochondrial state 3 respiration(ST3),state 4 respiration(ST4),respiratory control rate(RCR),the rate of oxidative phosphorylation(OPR),and oxygen consumption rate of the complex Ⅳ.MMP was detected by Rhodamine 123 method at the above-mentioned concentrations of NaCN.Results NaCNat0.01,0.1and0.25mmol/Linhibited the mito-chondrial oxidative respiratory function,and decreased MMP significantly.The inhibitory effects of NaCN onenergy metabolism in mitochondria was in a dose-dependent manner.Compared with the treatment of NaCN atthe corresponding concentration in the control group,mitochondrial function in the acute hypoxia group was in-hibited more seriously.Conclusion Acute hypoxia exacerbates the inhibitory effects of sodium cyanide on en-ergy metabolism in rat liver mitochondria.Its mechanism might be relevant to the decoupling of oxidative phos-phorylation,functional down-regulation of complexⅣin respiratory chain and changes of the mitochondrialmembrane potential in liver mitochondria from rats exposed to acute hypoxia.