ABSTRACT
Aloe vera, a popular succulent perennial medicinal plant with a wide range of phytochemicals that have shown various pharmacological activities including anti-oxidant, antimicrobial, antidiabetic, wound healing promotion and so on. Acemannan, aloe-emodin, aloin, aloesin, and emodin are widely investigated active constituents that show various pharmacological activities. Thus, the purpose of this review is to highlight previous pharmacological studied conducted in vivo, in vitro and human assays over the past decades. As current pharmacological research is focused on anticancer and neurological action, it would be interesting and important to study the main compounds present in Aloe vera for therapeutic purposes.
ABSTRACT
Objective:To screen small-molecule inhibitors of tyrosine kinase receptor B2 (EphB2) by using a molecular docking method, and to investigate their effect on cutaneous squamous cell carcinoma (CSCC) and possible mechanisms of action.Methods:The three-dimensional structure of EphB2 protein and its ligand binding sites were predicted by using the docking tool Schrodinger, and high-throughput virtual screening of EphB2 inhibitors was carried out by molecular docking. The anti-CSCC effect and mechanism of action of the screened EphB2 inhibitors kaempferitrin and aloe-emodin (AE) were verified in in vitro and in vivo experiments. In the in vitro experiments, human CSCC cell lines A431 and SCL-1, as well as the human immortalized keratinocyte HaCaT, were all divided into blank control group, dimethyl sulfoxide (DMSO) group, AE group and kaempferitrin group. Methyl thiazol tetrazolium (MTT) assay (AE at concentrations of 20, 40, 80, 160 μmol/L, kaempferitrin at concentrations of 12.5, 25, 50, 100 μmol/L), scratch and Transwell assays (AE at a fixed concentration of 80 μmol/L, kaempferitrin at a fixed concentration of 50 μmol/L) were performed to analyze the effect of EphB2 inhibitors on the proliferation, migration and invasion of CSCC cells. In the in vivo experiments, specific pathogen-free BALB/c female nude mice were subcutaneously injected with 0.2 ml of A431 cell suspension. After tumor growth, 24 tumor-bearing mice were randomly and equally divided into 4 groups: AE group and kaempferitrin group intraperitoneally injected with 20 mg·kg -1·d -1 AE and 25 mg·kg -1·d -1 kaempferitrin respectively, blank control group and DMSO group intraperitoneally injected with the same volume of sodium chloride physiological solution and DMSO respectively; the tumor size and body weight of nude mice were measured weekly; after consecutive treatment for 28 days, transplanted tumors were resected from the nude mice for hematoxylin and eosin (HE) staining, and real-time fluorescence-based quantitative PCR (qRT-PCR) and Western blot analysis were performed to analyze the effect of AE and kaempferitrin on the mRNA and protein expression of E-cadherin, vimentin, glycogen synthase kinase 3β (GSK-3β), phosphorylated GSK-3β (p-GSK-3β) and β-catenin respectively. One-way analysis of variance and t test were used for comparisons between groups. Results:Two small-molecule compounds AA-504/20999031 (kaempferitrin) and AA-466/21162055 (AE) with high inhibitory activity against EphB2 were screened out. MTT assay showed that both AE and kaempferitrin exhibited strong cytotoxicity to SCL-1 and A431 cells compared with HaCaT cells, and their toxicity increased with the increase of their concentration ( F = 17.95, 11.34, respectively, both P < 0.001) ; after 48-hour treatment, the 50% inhibitory concentrations (IC50s) of AE against SCL-1 and A431 cells were 124.59 and 80.85 μmol/L respectively, and the IC50s of kaempferitrin against SCL-1 and A431 cells were 119.64 and 64.96 μmol/L respectively. Scratch assay showed that the migration distance of A431 cells was significantly shorter in the AE group and kaempferitrin group (36.7 ± 1.0 μm, 44.7 ± 3.5 μm, respectively) than in the DMSO group (88.1 ± 1.4 μm, F = 52.34, P < 0.001), while there was no significant difference in the migration distance of HaCaT cells among the above groups ( F = 1.73, P = 0.238). Transwell assay showed that the number of A431 cells crossing the Transwell membrane significantly decreased in the AE group and kaempferitrin group (145.0 ± 2.5, 94.7 ± 4.1, respectively) compared with the DMSO group (195.3 ± 5.7, F = 72.85, P < 0.001), while neither AE nor kaempferitrin showed significant inhibitory effects of on the number of HaCaT cells crossing the Transwell membrane ( F = 3.91, P = 0.055). The animal experiment revealed significantly decreased volumes of transplanted tumors in nude mice in the AE group and kaempferitrin group (407.42 ± 70.37 mm 3, 368.77 ± 62.7 mm 3, respectively) compared with the DMSO group (841.88 ± 84.63 mm 3, F = 73.78, P < 0.001). HE staining confirmed that AE and kaempferitrin could improve pathological changes of transplanted tumors. qRT-PCR and Western blot analysis showed that AE and kaempferitrin significantly up-regulated the mRNA and protein expression of E-cadherin and p-GSK-3β in tumor tissues (all P < 0.001), and down-regulated the mRNA and protein expression of vimentin, β-catenin and GSK-3β (all P < 0.001) . Conclusion:The small-molecule inhibitors screened by molecular docking can form a stable complex with EphB2, and inhibit the progression of CSCC by affecting the Wnt/β-catenin pathway-induced epithelial-mesenchymal transition.
ABSTRACT
ObjectiveTo study the inhibitory effect of aloe-emodin （AE） on aluminum ion （Al3+）-induced β-amyloid protein 42 （Aβ42） aggregation and its depolymerization on formed Aβ42-Al3+ aggregates in vitro， and to investigate the effect of AE on the cytotoxicity of Aβ42 aggregation in the presence of Al3+. MethodThe Aβ42 group， Aβ42+Al3+ group， Aβ42+AE group， Aβ42+Al3++AE group and the depolymerization test group were set up in the experiment. The aggregation fibrosis process， aggregation morphology， aggregation size and cytotoxicity of Aβ42 in each group were detected by thioflavin T （ThT） fluorescence assay， transmission electron microscopy （TEM）， dynamic light scattering （DLS） experiment and thiazolyl blue （MTT） cytotoxicity assay. ResultCompared with the Aβ42 group， Al3+ could promote Aβ42 aggregation， increase the fluorescence intensity of ThT by 124.48%， induce the aggregation of Aβ42 to form fiber bundles with larger particle size， and significantly reduce the cell viability of human neuroblastoma SH-SY5Y cells （P<0.01）， thus reducing the cell survival rate to 51.05%. AE not only inhibited Aβ42 aggregation， but also inhibited Al3+-induced Aβ42 aggregation in a concentration-dependent manner. Compared with the Aβ42+Al3+ group， high concentration of AE could reduce the ThT fluorescence intensity to 41.66%， and change the polypeptide aggregation pathway to form amorphous aggregates with small particle size. Besides， it significantly inhibited the cytotoxicity of Aβ42 induced by Al3+ （P<0.01）， and restored the cell survival rate to 84.87%. Further depolymerization was conducted， AE could depolymerize Aβ42-Al3+ aggregates to make the formed aggregates disappear and form some small-particle short fibers and amorphous structure aggregates with low toxicity. ConclusionAE can inhibit Aβ42 aggregation and cytotoxicity in the presence of Al3+， depolymerize the formed Aβ42-Al3+ aggregates and alleviate the cytotoxicity， thus laying the foundation for exploring the mechanism of AE in the treatment of Alzheimer's disease.
ABSTRACT
ObjectiveTo study the inhibitory effect of aloe-emodin （AE） on aluminum ion （Al3+）-induced β-amyloid protein 42 （Aβ42） aggregation and its depolymerization on formed Aβ42-Al3+ aggregates in vitro， and to investigate the effect of AE on the cytotoxicity of Aβ42 aggregation in the presence of Al3+. MethodThe Aβ42 group， Aβ42+Al3+ group， Aβ42+AE group， Aβ42+Al3++AE group and the depolymerization test group were set up in the experiment. The aggregation fibrosis process， aggregation morphology， aggregation size and cytotoxicity of Aβ42 in each group were detected by thioflavin T （ThT） fluorescence assay， transmission electron microscopy （TEM）， dynamic light scattering （DLS） experiment and thiazolyl blue （MTT） cytotoxicity assay. ResultCompared with the Aβ42 group， Al3+ could promote Aβ42 aggregation， increase the fluorescence intensity of ThT by 124.48%， induce the aggregation of Aβ42 to form fiber bundles with larger particle size， and significantly reduce the cell viability of human neuroblastoma SH-SY5Y cells （P<0.01）， thus reducing the cell survival rate to 51.05%. AE not only inhibited Aβ42 aggregation， but also inhibited Al3+-induced Aβ42 aggregation in a concentration-dependent manner. Compared with the Aβ42+Al3+ group， high concentration of AE could reduce the ThT fluorescence intensity to 41.66%， and change the polypeptide aggregation pathway to form amorphous aggregates with small particle size. Besides， it significantly inhibited the cytotoxicity of Aβ42 induced by Al3+ （P<0.01）， and restored the cell survival rate to 84.87%. Further depolymerization was conducted， AE could depolymerize Aβ42-Al3+ aggregates to make the formed aggregates disappear and form some small-particle short fibers and amorphous structure aggregates with low toxicity. ConclusionAE can inhibit Aβ42 aggregation and cytotoxicity in the presence of Al3+， depolymerize the formed Aβ42-Al3+ aggregates and alleviate the cytotoxicity， thus laying the foundation for exploring the mechanism of AE in the treatment of Alzheimer's disease.
ABSTRACT
Objective: A method was established to obtain fingerprint and determination of ninecomponents in rhubarb based on ultra-performance liquid chromatography photodiode array detection (UPLC-PDA), and 17 batches of rhubarb from different regions, different varieties and different growth years were analyzed. Methods: A ThermoSyncronis C18 column (100 mm × 2.1 mm, 1.7μm) was used with a gradient of acetonitrile (A)-0.1% formic acid (B) as a mobile phase. Fingerprint data was imported intoSIMCA-P 14.1 software for cluster analysis and principal component analysis. At the same time, a total of ninecomponents including sennoside B, rhein-8-O-β-D-glucoside, sennoside A, emodin-8-O-β-D-glucoside, aloe-emodin, rhein, emodin, chrysophanol, physcion were quantitatively analyzed. Results: 20 common peaks were found in the fingerprints of 17 batches of rhubarb, and 9 peaks were identified by standard compounds. Cluster analysis and principal component analysis showed that Rheum tanguticumwas similar to Rheum officinaleand could be distinguished with Rheum palmatumwell. Fouryears and fiveyears of R. tanguticum could not be distinguished, oneyear and twoyears of R.palmatumcould not be distinguished neither. The determination of the indicator components showed thatR. tanguticum was higher than the other two kinds of rhubarb; Fouryears of R. tanguticum was better than five years, and twoyears of R.palmatumwas better than oneyear. Conclusion: This method established rhubarb fingerprint combined with multi-component determination based on UPLC-PDA technology could quickly, scientifically and accurately distinguish rhubarb of different origins. The preliminary evaluation of the rhubarb in different years and a basis for distinguishing the source of rhubarb was also provided.
ABSTRACT
Objective: To establish an ultra-high performance liquid chromatography coupled with hybrid quadrupole-orbitrap mass spectrometry (UPLC-ESI-HRMS) determination method for simultaneous determination of 11 components from raw, wine-broiled, carbon-fried and steamed Rheum pumilum roots. Methods: The chromatographic separation was performed on a KinetexTM C18 column (150 mm × 4.6 mm, 2.6 μm) with a gradient elution of acetonitrile and 0.1% formic acid in water at flow rate 0.3 mL/min, the injection volume was 1 μL and column temperature was 32 ℃. The mass spectrometry was detected using ESI ion source and negative ion mode. Results: Eleven components gallic acid, epicatechin, polydatin, rhaponticin, luteolin, emodin-8-O-β-D-glucoside, aloe-emodin, rhein, emodin, chrysophanol and physcion showed a good linear relationship within a certain concentration range. The precision, repeatability and stability of the method were good for the determination of 11 components. The average recoveries varied between 91.31% and 107.08% and the RSD were between 1.73% and 3.58%. The content of gallic acid, polydatin, emodin-8-O-β-D-glucoside and emodin changed in processsed products of R. pumilum roots. The content of gallic acid and emodin increased significantly in the wine-broiled and carbon-fried process. The content of emodin-8-O-β-D-glucoside in the carbon-fried process was significantly reduced, and the content of polydatin was significantly reduced in all processed products. Conclusion: This determination method is simple, stable, accurate and reliable. It can be applied for rapid quantitative determination of 11 components in raw and processsed products of R. pumilum, which laid the foundation for further research on R. pumilum roots.
ABSTRACT
Objective: To study the relationship between the quantitative color value of the powder and the known component content of rhubarb charcoal, and lay the foundation for the establishment of the rhubarb charcoal processing process control and endpoint judgment based on the color quantitative value. Methods： Rhubarb charcoal samples were prepared at different temperatures and time. Based on the empirical judgment of the rhubarb charcoal processing, the visual analyzer and UV-Vis were used to quantify the color of the pieces and powder of rhubarb charcoal under different processing conditions. At the same time, the HPLC fingerprint method was used to evaluate the dynamic changes of chemical components during the processing of rhubarb charcoal, and the quantitative value of the color of the sample during the processing of rhubarb charcoal was correlated with the characteristic components of the HPLC fingerprint using the multivariate statistical method. Results: During the processing of rhubarb charcoal, as the degree of carbonization increased, the apparent color of the sample changed from light yellowish brown to burnt black. There was a high correlation between the lightness value (L*), red-green value (a*) of the sample pieces and powder and the yellow and blue values (b*). The area of the 26 characteristic peaks had varying degrees of correlation with the chromaticity value. Among the 14 known components, five bound anthraquinones (aloe-emodin-8-O-β-D-glucoside, rhein-8-O-β-D-glucoside, chrysophanol-8-O-β-D- glucoside, emodin-8-O-β-D-glucoside, physcion-8-O-β-D-glucoside), and two sennosides (sennoside A, sennoside B) had a linear positive correlation with the chromaticity value. The content of five free anthraquinones (aloe-emodin, rhein, chrysophanol, emodin, and physcion), gallic acid and 5-hydroxymethylfurfural (5-HMF), whose contents increased first and then decreased, showed a quadratic correlation with the chromaticity value. Conclusion: The subjective judgment of rhubarb charcoal in the processing process is consistent with the quantitative color value analysis. The quantitative color value has a clear correlation with the content of 14 active chemical components. It is preliminarily inferred that the color quantitative value can be used as the quality of the rhubarb charcoal processing process control and end-point determination indicators to achieve efficient and rapid identification of the processing quality of rhubarb charcoal, which can provide new ideas for the monitoring and quality control of the rhubarb charcoal processing process.
ABSTRACT
Objective: To explore the potential mechanism of Xiahuang Granules in the treatment of opioid-induced constipation. Methods: Various medicinal ingredients and targets information of Xiahuang Granules were found in TCMSP database. In Genecards database, "opiod constipation", "opioid-induced bowel dysfunction" and "opioid-induced constipation" were used as keywords to search for targets related to opioid-induced constipation, and the active targets mapping of Xiahuang Granules were selected as the research targets. The common targets were imported into the STRING database to build the targets interaction network diagram, and Cytoscape 3.3.0 software was used for visual processing to screen out the core targets. The OmicsBean analysis platform and STRING database were used to conduct GO function enrichment and KEGG pathway enrichment analysis on the targets. Results: A total of 55 chemical constituents, 158 candidate target genes, 86 common targets after mapping Venny, 49 corresponding chemical components, 12 core targets and 19 main chemical components of Xiahuang Granules were obtained by screening. GO functional enrichment analysis showed 4 150 biological process items, involving chemical stimulus cell reactions, chemical reactions, biological quality control and other processes; A total of 302 cell composition items, involving voxel projection, extracellular space, and whole membrane processes; A total of 459 molecules function items, involving processes such as protein binding, molecular transduction activity, and enzyme binding were obtained. KEGG enrichment analysis revealed 149 signaling pathways related to the effect of Xiahuang Granules, involving the AGE-RAGE signaling pathway in diabetic complications and tumor necrosis factor signaling pathway, etc. The network of "medicinal herb-component-target-pathway" of Xiahuang Granules was established. Conclusion: The main chemical components of Xiahuang Granules including naringenin, nobiletin, aloe emodin, rhein may regulate endocrine resistance and tumor necrosis factor signaling pathways by acting on key proteins such as TNF, MAPK3, IL-6, VEGFA, and PTGS2, thus play a role in laxative, antispasmodic, and promoting gastrointestinal motility, which provides theoretical basis for Xiahuang Granules to treat opioid-induced constipation and is consistent with the preliminary verification results of Xiahuang granules.
ABSTRACT
The herbal formula, DF-02, consisting of Ephedra intermedia and Rheum palmatum are used for the treatment of the metabolic diseases such as obesity and liver fibrosis in Korean local clinics. We aimed to develop the simultaneous analytical conditions for four standards, (+)-pseudoephedrine (PSEP) and (−)-ephedrine (EP) for E. intermedia, and aloe-emodin (AE) and chrysophanol (CP) for R. palmatum using HPLC-UV techniques. The validated conditions yielded the high precision (relative standard deviation (RSD) 0.9994). As a result, four standards of DF-02 were simultaneously determined under the developed method, which will be utilized for the quality control or evaluation of DF-02 and many herbal preparations containing E. intermedia and R. palmatum.
Subject(s)
Calibration , Chromatography, High Pressure Liquid , Ephedra , Liver Cirrhosis , Metabolic Diseases , Methods , Obesity , Plant Preparations , Quality Control , RheumABSTRACT
Objective:To study nephrotoxicity induced by long-term administration of different doses of aloe-emodin in mice, and explore its mechanism. Method:A total of 30 male and female Kunming mice were randomly divided into normal control group, and low-dose aloe-emodin group,high-dose aloe-emodin group (0.8,1.6 g·kg-1). Every dose of group was administered intragastrically for 11 weeks,twice daily. effect of serum urea nitrogen (BUN),creatinine (SCr),superoxide dismutase (SOD),malondialdehyde (MDA),Glutathione (GSH/GSSG) and Glutathione Peroxidase (GSH-Px) levels were detected by biochemical kits according to manufacturer's instruction. Enzyme-linked immune assay was used to determine serum tumor necrosis factor (TNF)-α and interleukins(IL)-6 levels. Hematoxylin eosin (HE) staining was used to detect renal pathological changes in kidney tissues, and cysteine aspartic acid specific protease(Caspase)-3 and transforming growth factor(TGF)-β1 proteins were determined by immunohistochemistry. Result:According to results,compared with normal control group,the levels of BUN and SCr in serum with high-dose aloe-emodin were increased. The renal tubules in low-dose group were mildly injured,while renal tubules and glomeruli of high-dose group were moderately damaged. Compared with normal control group,the level of SOD was significant decreased (PPPPα and IL-6 were increased,the expression of TGF-β1 protein in kidneys was increased in low-dose and high-dose groups (PConclusion:results show that 1.6 g·kg-1 aloe-emodin was administered intragastrically for 11 weeks,which had toxic effects on kidney in mice. The mechanism may be related to oxidative stress,apoptosis and TGF-β1 protein expression.
ABSTRACT
Objective To compare the changes of 17 constituents in rhubarb before and after the processing of honeyed wine. Methods HPLC method was used to quantitatively analyze 17 components including gallic acid, catechin, epicatechin, polydatin, ferulic acid, sennoside B, rhein-8-O-β-D-glucoside, sennoside A, emodin-1-O-glucoside, chrysophanol-8-O-glucoside, kaempferol, emodin-8-O-glucoside, aloe-emodin, rhein, emodin, chrysophanol, and physcion in honeyed wine rhubarb. Results Seventeen kinds of ingredients, gallic acid, catechin, epicatechin, polydatin, ferulic acid, sennoside B, rhein-8-O-β-D-glucoside, sennoside A, emodin-1-O-glucoside, chrysophanol-8-O-glucoside, kaempferol, emodin-8-O-glucoside, aloe-emodin, rhein, emodin, chrysophanol, and physcion, were well separated. The linear ranges of which were 25.94—830.00 μg/mL (r = 0.999 1), 28.20—1 805.00 μg/mL (r = 0.999 8), 77.03—2 465.00 μg/mL (r = 0.999 2), 25.00—1 600.00 μg/mL (r = 0.999 3), 11.41—730.00 μg/mL (r = 0.999 6), 7.85— 1 005.00 μg/mL (r = 0.999 0), 210.47—6 735.00 μg/mL (r = 0.999 0), 113.28—3 625.00 μg/mL (r = 0.999 6), 10.94—700.00 μg/mL (r = 0.999 8), 218.80—1 415.00 μg/mL (r = 0.999 6), 55.00—1 760.00 μg/mL (r = 0.999 2), 48.44—1 550.00 μg/mL (r = 0.999 7), 19.22—625.00 μg/mL (r = 0.999 6), 18.91—1 210.00 μg/mL (r = 0.999 1), 14.06—450.00 μg/mL (r = 0.999 2), 61.41—1 965.00 μg/mL (r = 0.999 4), and 25.16—805.00 μg/mL (r = 0.999 5), respectively. The averages of the total recovery were 93.71%—102.77% at the three concentrations of 17 components. The content of sennoside and anthraquinone was significantly reduced after rhubarb was processed by Yi method, while the content of gallic acid was significantly increased. Conclusion For the first time, HPLC method was used to quantitatively analyze the 17 components in honeyed wine rhubarb, which provided a reference for the quality standards of honeyed wine rhubarb.
ABSTRACT
Objective: A method for simultaneous determination of 10 active ingredients in Dahuang Lidan Tablets (DLP) by HPLC wavelength switching method was established, and its quality was evaluated by statistical analysis. Methods: A Phenomenex Kinetex C18 column was used with a column temperature of 30 ℃ and a mobile phase gradient of methanol-0.15% phosphoric acid. The flow rate was 1 mL/min and the detection wavelengths were 265.0 nm (0-5.8 min, gallic acid), 283.9 nm (5.8-7 min, 5-hydroxymethyl furfural), 222.2 nm (7-18 min, corilagin, p-hydroxybenzaldehyde), 256.7 nm (18-74 min, ellagic acid, aloe-emodin, rhein, emodin, chrysophanol, emodin methyl ether), respectively. Statistical analysis of the content of components in 10 batches of drugs was performed using SPSS 21 Software. Results: The linearity of 10 components in the respective mass concentration ranges was good (r > 0.998 0), the average sample recovery was in the range of 98.45%-100.12%, and the RSD was in the range of 0.80%-2.51%. The content of 10 components was as follow: gallic acid (8.371-11.438 mg/tablet), 5-hydroxymethylfurfural (0.046-0.087 mg/tablet), corilagin (0.721-2.094 mg/tablet), p-hydroxybenzaldehyde (0.034-0.065 mg/tablet), ellagic acid (1.736-1.996 mg/tablet), aloe-emodin (0.337-0.440 mg/tablet), rhein (1.636-2.562 mg/tablet), emodin (0.602-0.846 mg/tablet), chrysophanol (0.388-0.566 mg/tablet) and emodin methyl ether (0.621-0.781 mg/tablet). The quality of the 10 batches of samples was basically the same. Conclusion: This method is simple and accurate and can be used for the quality control of DLP.
ABSTRACT
Objective:To establish an HPLC fingerprint of Yiqing granules to provide reference for the effective quality control. Methods:The analysis was carried out on an analytical Agilent ZORBAX SB-C18(250 mm×4.6 mm,5 μm)column with gradient elution by acetonitrile(A)-0.2% phosphoric acid solution(adjusting pH to 3.0 with triethylamine,B)(0-5 min:10% A→20%;5-20 min:20% A→30% A;20-25 min:30% A→45% A;25-35 min:45% A;35-45 min:45% A→80% A;45-50 min:80% A)at the detection wavelength of 254 nm and the flow rate of 1.0 ml·min-1,and the column temperature was 35 ℃. Twelve batches of Yiqing granules were determined by HPLC and a common mode of fingerprint was established. Results:There were 22 common peaks in the fingerprints of the twelve batches of Yiqing granules,and seven of them were identified. The similarities of fingerprints of the twelve batches of Yiqing granules were over 0.990 suggesting good reproducibility. Conclusion:The developed method is accurate and feasible,and can be used for the quality control of Yiqing granules.
ABSTRACT
Objective:To optimize the decoction process of Xiaochengqi decoction by Box-Behnken response surface method. Methods:Rhein,aloe-emodin and synephrine were chosen as the evaluation indices. After the material-liquid ratio, decocting frequency and decocting time were studied,Box-Behnken response surface method was used to optimize the three factors using the weighted calculation value of the extraction rate of above indicator components. Results:The best conditions were as follows:the material-liquid ratio was 1:10,the decocting frequency was 3,and the decocting time was 1.0 h. The deviation of measured value and predicted value was 1.15%. Conclusion:The best decocting conditions are promising and reliable.
ABSTRACT
Objective To establish a quantitative analysis method of multiple active components (gallic acid, tanshinol, ferulic acid, rosmarinic acid, salvianolic acid B, luteolin, apigenin, aloe-emodin, emodin, butylidenephthalide, and levistilide A) in Fukejing Capsule (FC) based on ultra performance liquid chromatography-quadrupole/orbitrap high resolution mass spectrometry (UPLC-Q- Orbitrap HRMS). Methods The column was BEH C18 (50 mm × 2.1 mm, 1.7 μm) and the mobile phase was consisted of acetonitrile-water (containing 0.1% formic acid) at a flow rate of 0.2 mL/min with gradient elution; Mass spectrometer conditions: Heated electrospray ionization source (HESI) and full mass quantify methods were used to perform the determination, and the results of the contents were imported into the muti-data processing soft ware SIMCA14.0 to make a quality evaluation. Results Under the optimized conditions, gallic acid, tanshinol, ferulic acid, rosmarinic acid, salvianolic acid B, luteolin, apigenin, aloe-emodin, emodin, butylidenephthalide, and levistilide A all showed good liner relationship (r ≥ 0.999 5) in the range of 0.15—1.50, 0.80—8.00, 1.50—15.00, 0.04—0.40, 0.015—0.150, 0.10—1.00, 0.004—0.040, 0.004—0.040, 0.02—0.20, 0.01—0.10, and 0.04—0.40 μg/mL, respectively; The results of the accuracy, the repeatability and the stability all reached the standards (RSD ≤ 4%); The recoveries ranged from 98%—102% and RSDs were below 3%; The result of the content ranges in different batches were 26.36—31.03 μg/g (gallic acid), 178.85—210.79 μg/g (tanshinol), 320.91—343.16 μg/g (ferulic acid), 2.84—3.09 μg/g (rosmarinic acid), 3.55—4.25 μg/g (salvianolic acid B), 27.33—32.36 μg/g (luteolin), 1.89—2.13 μg/g (apigenin) A, 0.47—0.60 μg/g (aloe-emodin), 3.17—3.57 μg/g (emodin), 1.99—2.54 μg/g (butylidenephthalide), and 7.51—8.53 μg/g (levistilide A). The analysis results showed that the quality of the most batches was stable, the tanshinol and ferulic acid had a great influence on the quality of the medicine, and could be monitored to ensure the quality of different batches. Conclusion The methods established in this paper have a high sensitivity and accuracy; The results of the methodology conform to the relevant requirements and the methods can rapidly determinate the multiple active components in FC; The research also provides a new scientific basis and reference for the quality assessment at the same time.
ABSTRACT
Objective To study the chemical constituents of Lianhua Qingwen Capsule (LQC). Methods The compounds were isolated and purified by column chromatography over silica gel and sephadex LH-20, and preparative RP-HPLC. Their structures were elucidated by physicochemical properties and spectral analyses. Results Twenty compounds were isolated and identified as aloe-emodin (1), phillygenol (2), cepharanone B (3), pinoresinol (4), epipinoresinol (5), pinoresinol monomethyl ether (6), piperolactam A (7), (E)-ethyl 3-(4-hydroxyphenyl) acrylate (8), formononetin (9), isoliquiritigenin (10), naringenin (11), kaempferol (12), citreorosein (13), rhein (14), physcion-8-O-β-D-glucopyranoside (15), 4,5-dioxodehydroasimilobine (16), kaempferol-3-O-α- L-ramnopyranosyl-4″-O-E-(4-hydroxy)-cinnamoyl (17), chrysophanol-1-O-β-D-glucopyranoside (18), chrysophanol-8-O-β-D- glucopyranoside (19), and emodin-8- O-β-D-glucopyranoside (20). Conclusion Compounds 2, 3, 5-17, 19, and 20 are isolated from LQC for the first time. This study establishes the foundation for explaining the efficient substance of LQC.
ABSTRACT
Objective: To study the chemical constituents of Lianhua Qingwen Capsules. Methods: The compounds were isolated and purified by gel column chromatography, MPLC, and preparative HPLC from 50% ethanol fraction of macroporous resin column chromatography of Lianhua Qingwen crude extracts. Their structures were elucidated by the spectral analyses. Results: Eight compounds were isolated and identified as 10-O-(p-hydroxycinnamoyl)-adoxosidic acid (1), aloe-emodin-8-O-β-D-glucopyranoside (2), quercitrin (3), matairesinol-4’-O-β-D-glucoside (4), liquiritin apioside (5), epi-vogeloside (6), vogeloside (7), and caffeic acid ethyl ester (8). Conclusion: Compound 1 is a new compound named lianhua iridoid A. Compounds 5-8 are isolated from Lianhua Qingwen Capsules for the first time. This study provides substance foundation for chemical research of Lianhua Qingwen Capsules.
ABSTRACT
Objective The contents of sennoside A, sennoside B, echinacoside, verbascoside, hesperidin, aloe-emodin, rhein, emodin, chrysophanol, and rheochrysidin in Paidu Yangyan Capsules (PYC) were determined by HPLC. The quality of different batches of psoralen were evaluated by chemical pattern recognition. Methods HPLC was established for the simultaneous determination of 10 components in 15 batches of PYC with variable wavelength detector, and the results were analyzed by principal component analysis and cluster analysis to comprehensively evaluate the difference in quality of PYC which is available in the market. Results The contents of sennoside A, sennoside B, echinacoside, verbascoside, hesperidin, aloe-emodin, rhein, emodin, chrysophanol, and rheochrysidin in PYC had good linear relationship in the ranges of 0.167 4—2.093 0, 0.093 5—1.177 0, 0.075 9—0.948 6, 0.049 9—0.623 7, 0.109 0—1.363 0, 0.033 5—0.418 4, 0.079 7—0.996 1, 0.013 9—0.174 3, 0.025 2—0.315 6, and 0.012 3—0.061 4 μg/mL, the average sample recovery rate range were 99.84%, 99.15%, 99.34%, 99.81%, 100.6%, 99.36%, 99.86%, 100.1%, 100.1%, and 99.76% (RSD < 2.0%), the detection limits were 0.64—2.18 ng/mL, quantitative limits were 3.19—9.55 ng/mL, the content of 10 active ingredients in 15 batches of samples respectively were 0.712 7—1.118 0, 0.403 9—0.522 0, 0.236 4—0.320 3, 0.671 1—1.183 0, 0.0580—0.1467, 0.108 7—0.2592, 0.0147—0.0501, 0.5173—0.5828, 0.2754—0.3051, and 0.1621—0.1853 mg/grain and there were some differences among the samples in different batches, mainly due to different rhubarb dosages. Conclusion The established method is simple, accurate and reproducible, and can be used to control the quality of PYC. It is suggested that the quality control of Rhei Radix et Rhizoma should be strengthened in the process of preparation production for providing a reference of PYC follow-up quality improvement.
ABSTRACT
Objective To investigate the effect of aloe-emodin on Akt/mTOR,apoptosis and migration of autophagy-related pathway in colorectal cancer LOVO cells. Methods The LOVO cells were divided into control group(normal culture),low dose group(LOVO cells+10 μmol/L aloe-emodin culture 30 min), medium dose group (LOVO cells + 30 μmol/L aloe-emodin culture 30 min) and high dose group (LOVO cells + 50 μmol/L aloe veratin culture 30 min). The cell proliferation rate was measured by CCk-8 method at 24,48 and 72 h respectively. The apoptotic rate of each group was detected by flow cytometry. The migration ability of each group was detected by Transwell chamber. Western blot was used to detect the expression of IC3-Ⅰ,IC3-Ⅱ,Beclin-1,mTOR,p-mTOR,Akt and p-Akt.Results Compared with that of the control group,the cell rate,migration ability and apoptosis rate of the low dose group was not significantly changed(P>0.05).The cell rate and migration ability of the medium and high dose groups was significantly decreased and the apoptosis rate was significantly increased:0.43 ± 0.03,0.59 ± 0.04 vs.0.16 ± 0.00;0.57 ± 0.07,1.06 ± 0.17 vs.0.34 ± 0.02;0.37 ± 0.02, 0.49 ± 0.01 vs.0.13 ± 0.00,P<0.05,there was dose dependent(P<0.05).Compared with those of the control group,the expressions of IC3-Ⅰ,IC3-Ⅱ,Beclin-1,mTOR,p-mTOR,Akt and p-Akt in the low dose group showed no significant change(P>0.05),those were decreased in middle and high dose groups which showed significant differences:0.85 ± 0.08,0.37 ± 0.02 vs.2.08 ± 0.07;1.42 ± 0.09,1.19 ± 0.02 vs.1.97 ± 0.11;0.97 ± 0.00,0.84 ± 0.06 vs.1.19 ± 0.02;0.43 ± 0.02,0.31 ± 0.01 vs.0.98 ± 0.08,P<0.05, there were dose dependent (P < 0.05). Conclusions Aloe-emodin can promote autophagy, apoptosis there inhibit the growth,migration of colorectal cancer LOVO cells by reducing the expression of Akt/mTOR signaling pathway and reducing its phosphorylation level.
ABSTRACT
Under the traditional processing theory "wine processing could promote the efficacy", Rhubarb after wine processing could treat the upper energizer diseases such as red swelling, and breath sores. Processing changes the medicinal properties of rhubarb, and thus results in different focuses in clinical application. In this study, a sensitive and specific method was developed for the determination of aloe-emodin, rhein and emodin in rats tissue. Rhubarb raw materials and its wine processed decoction were given to SD rats respectively by gavage administration, and then the contents of aloe-emodin, rhein and emodin in the tissues (heart, lung, brain, liver, kidney) were determined by HPLC-MS to explore the effect of wine processing on free anthraquinones in rat tissues. Experimental results showed that wine processing can significantly change the distribution of aloe emodin, rhein and emodin in rats in vivo, and the distribution of these components was increased in heart and lung tissues.There was no significant change of distribution in the liver and the kidney as compared with raw product group, and these three ingredients were not detected in the brain, indicating that aloe-emodin, rhein, emodin can not pass through the blood brain barrier.Therefore, wine processing had greater effect on distribution of free anthraquinones in rat tissues.This also verified the theory of traditional Chinese medicine, providing experimental basis for rhubarb processing mechanism.