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ObjectiveTo observe the effect of ethyl acetate extract of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis on high-fat diet-induced apolipoprotein E gene knockout （ApoE-/-） mice， and explore its mechanism of treating atherosclerosis by regulating intestinal flora. MethodThirty-two 8-week-old male ApoE-/- mice were randomly divided into model group， rosuvastatin group （10 mg·kg-1）， high-， low-dose groups of ethyl acetate extract of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis （75， 25 mg·kg-1）， with 8 mice in each group. Eight C57BL/6 mice were used as blank group. After 8 weeks of continuous administration， blood was taken to determine the blood lipid level. Enzyme-linked immunosorbent assay （ELISA） was used to detect the contents of related indexes in serum of mice. Hematoxylin-eosin （HE） staining was used to observe the formation of aortic plaque in mice. Cecal contents were collected and 16S rRNA amplicon sequencing was used to detect intestinal flora. ResultCompared with the blank group， the plaque area of the model group was significantly increased with inflammatory infiltration， the contents of triglyceride （TG）， total cholesterol （TC）， low-density lipoprotein cholesterol （LDL-C）， inflammatory factors and inducible nitric oxide synthase （iNOS） were increased， while the content of high-density lipoprotein cholesterol （HDL-C） was decreased. Compared with the model group， rosuvastatin group and high- and low-dose groups of ethyl acetate extract of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis could improve the deposition of aortic plaque， reduce the contents of TG， TC， LDL-C， inflammatory factors and iNOS， and increase the content of HDL-C. Compared with the blank group， the relative abundances of Firmicutes and Proteobacteria in the model group increased， while the relative abundance of Bacteroidetes decreased. Alpha and Beta diversity analysis showed that samples of each group could be significantly isolated， and the total number and abundance of intestinal flora species in the model group were low. Compared with the model group， ethyl acetate extract of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis could increase the relative abundance of beneficial bacteria and decrease the relative abundance of pathogenic bacteria. ConclusionEthyl acetate extract of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis was mainly composed of flavonoids， which can treat atherosclerosis by regulating the intestinal flora and improve the pathological changes in the aorta of ApoE-/- mice induced by high-fat diet. The mechanism may be related to its ability to reduce the level of inflammatory factors， improve antioxidant capacity and repair the disorder of intestinal flora structure.
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Objective: To investigate the osteoblastogenic activity of the ethyl acetate (EtOAc) extract of Smilax glabra Roxb roots and its major active compound astilbin. Methods: Astilbin was isolated from EtOAc extract using silica gel chromatography combined with fraction crystallization. Chemical structure of astilbin was determined by analysis of the spectroscopic data in comparison with the literature. MTT method was used to detect the toxicity. Alkaline phosphatase (ALP) activity was determined by the spectrophotometric method at 405 nm using p-nitrophenyl phosphate as a substrate. Calcium deposition was stained with alizarin red-S, distained with cetylpyridium chloride, and quantified at 562 nm. In silico model for astilbin-ALP interaction was analyzed using AutoDock 4.2.6. The changes in expression of osteoblast differentiation related genes were determined using quantitative real-time PCR. Results: Both the EtOAc extract and astilbin had no toxicity toward osteoblast MC3T3-E1 cells at 5.0, 10, 25, and 50 μg/mL. At 25 μg/ mL, they enhanced ALP activity and mineralization of osteoblasts up to 30% and 55% for the EtOAc extract and 22% and 41% for astilbin, respectively. Molecular docking analysis of astilbin-ALP interaction revealed Arg167, Asp320, His324, and His437 were key residues participating in hydrophobic interaction; meanwhile, His434 and Thr436 residues were involved in hydrogen bond formation in the active site of human tissue-nonspecific ALP. Moreover, the expression level of genes opn, col1, osx, and runx2 were up-regulated in astilbin treated samples with the fold changes as 2.2; 3.7; 4.1; 2.3, respectively at 10 μg/mL (P<0.05). Conclusions: The EtOAc extract and its major compound astilbin exhibit osteoblastogenic activity by up-regulating important markers for bone cell differentiation. It could be a new and promising osteogenic agent with dual actions for therapeutic applications.
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To explore the protective effect and mechanism of ethyl acetate extract from Bidens bipinnata on hepatocyte damage induced by endoplasmic reticulum stress. Tunicamycin was used to establish the damage model in L02 cells. Methyl thiazolyl tetrazolium(MTT) colorimetric assay was used to investigate the survival rate of ethyl acetate extract from B. bipinnata in L02 cells injury induced by endoplasmic reticulum stress; the protein expressions of endoplasmic reticulum stress-related molecule glucose regulated protein 78(GRP78), PKR-like ER kinase(PERK), eukaryotic initiation factor-2(eIF2α), activating transcription factor 4(ATF4), C/EBP homologous protein(CHOP), B-cell CLL/lymphoma 2(Bcl-2), Bal-2 associated X apoptosis regulator(Bax) were examined by Wes-tern blot. The expressions of the above proteins were also detected after endoplasmic reticulum stress inhibitor(4-phenyl butyric acid) and CHOP shRNA-mediated knockdowns were added. The expressions of GRP78, PERK, CHOP in L02 cells were observed by immunofluorescence method. The results showed that ethyl acetate extract from B. bipinnata could significantly increase the survival rate of L02 cell injury caused by endoplasmic reticulum stress in a dose and time-dependent manner(P<0.05 or P<0.01). The expression levels of GRP78, PERK, eIF2α, ATF4, CHOP and Bax in the drug treatment groups were significantly down-regulated(P<0.05 or P<0.01), while Bcl-2 was significantly up-regulated(P<0.01). After endoplasmic reticulum stress inhibitor and CHOP shRNA-mediated knockdowns were added, the expression levels of GRP78, PERK, eIF2α, ATF4, CHOP, Bax in the drug treatment groups were significantly down-regulated(P<0.01), whereas Bcl-2 was significantly up-regulated(P<0.01). Immunofluorescence results showed that the expressions of GRP78, PERK, CHOP were consistent with the Western blot method. In conclusion, ethyl acetate extract from B. bipinnata has a significant protective effect on the damage of L02 cells caused by endoplasmic reticulum stress. The mechanism may be related to the inhibition of endoplasmic reticulum stress and the down-regulation of apoptosis in cells through the PERK/eIF2α/ATF4/CHOP signaling pathway.
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Acetates , Apoptosis , Bidens , Endoplasmic Reticulum Stress , Hepatocytes , Transcription Factor CHOP/genetics , eIF-2 Kinase/geneticsABSTRACT
OBJECTIVE：To i nvestigate the spectrum-effect relationship of analgesic and anti-inflammatory effects of ethyl acetate extract from Zhuang medicine Stahlianthus involucratus from different habitats. METHODS ：Ten batches of S. involucratus from different habitats were used as samples to investigate the anti-inflammatory and analgesic activities of ethyl acetate extracts by xylene induced ear swelling test and acetic acid induced writhing test in mice. HPLC fingerprints of 10 batches of ethyl acetate extract from S. involucratus were established and their similarity was evaluated by using Similarity Evaluation System of TCM Chromatogram Fingerprint （2012 edition），and the common peaks were identified by comparison with the control. The spectrum-effect relationship of anti-inflammatory and analgesic effects of ethyl acetate extract from S. involucratus were analyzed on the basis of Pearson correlation coefficient （auricle swelling degree and writhing times in 15 min as pharmacodynamic indexes ）and Grey relational analysis （inhibition rate of ear swelling and analgesic rate as pharmacodynamic indexes ）. RESULTS ： batches of ethyl acetate extract from S. involucratus had obvious anti-inflammatory and analgesic effects ； inhibition rates of ear swelling in mice were 46.43％-55.16％，and the analgesic rates of mice were 45.56％-52.72％. A total of 18 common peaks were identified in 10 batches of samples ，andthe similarity between them and the control fingerprint was 0.994-0.997. Compared with substance control ，the pea ks 1，2 and 4 were identified as protocatechuic acid ， p-hydroxy- 0771-4953513。E-mail：liangjie1101@126.com benzoic acid and p-hydroxybenzaldehyde，respectively. Results of Pearson correlation analysis showed that peak 10 and peak 18 were significantly negative correlated with auricle swelling degree and writhing times in 15 min（r were values －0.853，－0.738，P values were 0.002，0.015，respectively）. Results of Gray correlation degree analysis showed that the correlation degree of 18 common peaks with inhibition rate of ear swelling and analgesic rate were all greater than 0.65；among them ，peaks 14，1（protocatechuic acid ），17，9，4（p-hydroxybenzaldehyde），2（p-hydroxybenzoic acid ）， 16，7 and 6 showed the relatively high correlation degree （correlation degree ＞0.7）；peak 1（protocatechuic acid ），17，14，9，16，2 （p-hydroxybenzoic acid ）and 4（p-hydroxybenzaldehyde）showed the relatively high correlation degree （correlation degree ＞0.7）. CONCLUSIONS：The ethyl acetate extract of S. involucratus show good anti-inflammatory and analgesic effects. Peak 1 （protocatechuic acid ），2（p-hydroxybenzoic acid ），10，14，17，18 may be its main active ingredients.
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Aims@#To evaluate the antibacterial efficacy of ethyl acetate extract of Aspergillus flavus IBRL-C8 against Gram-positive and Gram-negative bacteria.@*Methodology and results@#In this experiment, an endophytic fungus which identified as A. flavus IBRL-C8 was extracted using ethyl acetate and methanol, from Senna siamea, prior to in vitro antibacterial test on eight Gram-bacteria. The results were significantly more enunciated to the ethyl acetate extract since the Gram-bacteria signified 9.0 to 20.0 mm of inhibition zones on Muller Hinton Agar (MHA) during disc diffusion assay. Minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of the extract were ranged from 125-1000 µg/mL and 125-2000 µg/mL, respectively. Time-kill assay depicted the ethyl acetate extract of A. flavus IBRL-C8 exceptionally retarded methicillin-resistant Staphylococcus aureus (MRSA) and also manifested extended antibacterial activity. The maximum reduction in cell numbers occurred at 2MIC concentration (250 µg/mL) during the interval time of 16 h. The malformations noticed from microscopic observations where the transformation of structural annihilation from regular spherical morphology to non-spherical shape with an irregular surface and also disruption around the cell membrane when the MRSA treated with ethyl acetate extract of A. flavus IBRL-C8. @*Conclusion, significance and impact of study@#This study proposed the ethyl acetate extract of A. flavus IBRL-C8 as a potential antibacterial agent against MRSA infection, which can be useful in pharmaceutical application.
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Aspergillus flavus , Anti-Bacterial AgentsABSTRACT
Objective: To study the chemical constituents of ethyl acetate extracts from the root bark of Celastrusmonospermus. Methods: The compounds were separated and purified by repeated column chromatographic methods including silica gel, ODS and Sephadex LH-20, and the chemical structures of compounds were determined by spectral data analyses. Results: Twelve compounds were obtained and identified as polpunonic acid (1), p-hydroxybenzoic acid (2), 4-hydroxy-3-methoxybenzoic acid (3), salaspermic acid (4), 3-oxo-2β-hydroxyfriedelan-29-oic acid (5), celastrol (6), 3α,22β-dihydroxy-olean-12-en-29-oic acid (7), orthosphenic acid (8), 21-oxo-2α,3α,22β-trihydroxy-29-nor-friedelan-24,2â-lactone (9), 2,3-dioxo-6α,10-dihydroxy-24-nor-friedelan-4,7-dien-29-oic acid (10), 2α,3β,19α,23-tetrahydroxyolean-12-en-28-oic acid (11) and 2α,3β-dihydroxyolean-12-en-23,28,30-trioic acid (12). Conclusion: Compounds 1, 3, 7, 10-12are obtained from this plant for the first time, and compounds 3, 11, 12 of them were separated from the genus Celastrus for the first time.
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OBJECTIVE：To study inhibitory activity of different extracts of Dracocephalum moldavica to clinical pathogenic bacteria，and to excavate its possible antibacterial mechanism. METHODS ：After extraction by 65％ ethanol and extraction by petroleum ether ，dichloromethane，ethyl acetate ，n-butanol，the different polar fractions of D. moldavica were obtained . Taking Klebsiella pneumoniae ，Staphylococcus aureus and other clinical multiple resistant pathogens as objects，the diameter of inhibition zone of different extraction fractions was measured by paper diffusion method ，and the antibacterial active fraction was screened. The minimal inhibitory concentration （MIC）of antibacterial active fraction to common clinical pathogens was determined by agar dilution method ；the growth curve of MRSA was drawn by turbidimetric method. The differentially expressed protein between antibacterial active fraction group and control group was screened by PEAK ® Q 8.5 software，and the gene ontology （GO）analysis and KEGG signaling pathway enrichment analysis were carried out by Blast 2 GO and KOBAS 3.0 online software. RESULTS ： Petroleum ether ，dichloromethane，ethyl acetate and n-butanol fraction of D. moldavica had no significant inhibitory effect on Gram-negative bacteria. The ethyl acetate fraction of D. moldavica had antibacterial activity in varying degrees against several kinds of Gram-positive bacteria as Staphylococcus aureus ，S. epidermidis；the diameter of inhibition zone was 10-16 mm，which was the active fraction. MICs of ethyl acetate fraction to S. aureus ，S. epidermidis and S. hominis were all 0.781 3 mg/mL；MIC to S. saprophyticus was 0.390 7 mg/mL；MICs to S. saprophyticus and standard strain of S. aureus were both 1.562 5 mg/mL. The 1， 2 times MIC of ethyl acetate could inhibit the growth of MRSA ，and the inhibitory activity increased with the increase of dose. A total of 300 differentially expressed proteins were screened （P＜0.01），of which 239 were up-regulated and 61 were down-regulated. The differentially expressed proteins were （No.81260478，No.816- mainly concentrated in cell sites such as cells ，cellular com- 60048） ponent，etc.，and in metabolic process such as cell process ， biological process and molecular functions such as catalytic activity，protein binding ，etc. They were mainly concentratedwang.zhanli@hotmail.com in microbial metabolism in different environments ，fructose and mannose metabolism signaling pathway （P＜0.05）. CONCLUSIONS ：The ethyl acetate fraction of D. Moldavica is the antibacterial active fraction ，and its activity may be related to the microbial metabolism and cell glycometablism.
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Objective:To investigate the chemical constituents from the ethyl acetate extract of 95% ethanol extract from Ajuga ovalifolia var. calantha. Method:A. ovalifolia var. calantha was percolated with 95% ethanol，and the percolate was concentrated under reduced pressure to obtain the extract. The extract was then dispersed with water and extracted with petroleum ether to obtain the petroleum ether extract fraction and water layer. Then ethyl acetate was used to extract the water layer to obtain the ethyl acetate extract fraction. The Compounds from the ethyl acetate extract fraction were isolated and purified by silica gel，Sephadex LH-20，MCI column，ODS column chromatography and semi-preparative high performance liquid chromatography. Their structures were elucidated by interpretation of NMR，ESI-MS and other spectral evidence. Result:17 Compounds were isolated and elucidated as bakkenolide-E（1），loliotide（2），isololiotide（3），（E）-linalool-1-oic acid（4），umbelliferone（5），phillygenin（6），1-（4-hydroxyphenyl）-ethanol（7），（2E）-8-hydroxy-2，6-dimethyl-2-octenoic acid（8），1H-indole-3-carboxylic acid（9），ajugarin I（10），ajugalactone（11），luteolin（12），20-hydroxyecdysone-2-acetate（13），benzyl-4'-hydroxy-benzoyl-3'-O-β-D-glucopyranoside（14），harpagide（15），6-deoxy-8-acetylharpagide（16），and acteoside（17）. Conclusion:Compounds 1，3-4，6-9，14 were isolated from the plants of Ajuga for the first time，and compounds 2，5，10，13，15-16 were isolated from this plant for the first time.
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To study the influence of ethyl-acetate extract of Polygonum amplexicaule var. sinense (EAEP) on lung cancer cellular proliferation and apoptosis and the underlying mechanisms. Methods The cell growth of A549 and H1299 cells was measured by cell counting kit-8 experiment and apoptosis was analyzed by flow cytometry. The treated cellular total mRNAs and proteins related to the apoptosis of A549 and H1299 cells were extracted, and then used for qRT-PCR and Western blotting experiments. Results The apoptosis rate of A549 and H1299 cells was observed to be markedly promoted by EAEP, and flow cytometry statistics suggested that this effect may mainly be produced by its pro-apoptotic procedures. qRT-qPCR and Western blotting assays also proved that EAEP significantly promoted expression of Bax and Caspase-3, and inhibited expression of Bcl-2. Conclusion EAEP may contain components that play a role in regulating cellular growth via influencing Bax and Bcl-2 pathway.
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Objective: To intensively study the chemical constituents from the seed cake of Camellia oleifera and its pharmacological activities,in order to provide scientific basic for its further development and utilization. Method: All kinds of column chromatography and spectral methods were employed to isolate and identify the monomeric compounds from its ethyl acetate portion of ethanol extract. The in vitro anti-inflammatory effects were evaluated by LPS-induced inflammatory model in RAW264.7 macrophages. Result: Eight phenolic acids and two flavonoids were isolated from the ethyl acetate soluble portion and identified as p-hydroxybenzoic acid(1),protocatechuic acid(2),gallic acid(3),methyl gallate(4),ethyl gallate(5),isovanillic acid(6),ethyl 3,4-dihydroxylbenzoate(7),2-(3',4'-dihydroxyphenyl)-1,3-benzodioxole-5-aldehyde(8),quercetin(9),rutoside(10). Among them, compounds 4-8 were first isolated from this plant. These compounds had good anti-inflammatory activities against NO production in LPS-stimulated RAW264.7 macrophages in an obvious dose-dependent manner. Among them, compound 8 showed a strongest activity. Conclusion: The above results show that the phenolic acids and flavonoids from seed cake of C. oleifera have good prospects for the development and application of anti-inflammatory drugs.
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OBJECTIVE： To study the chemical constituents in the ethyl acetate extract of Balanophora involucrate， and to provide reference for further enriching chemical constituent of the plant and the development and utilization of B. involucrate. METHODS： The whole plant of B. involucrate was extracted with 75％ ethyl alcohol. The extraction was carried out by petroleum ether， dichloromethane， ethyl acetate and n-butyl alcohol in turn. The chemical compounds from ethyl acetate extract part were isolated and purified by silica gel column， gel column and semi-preparative HPLC. The structures were identified on the basis of spectral spectrum （mass spectrum， hydrogen spectrum and carbon spectrum） data and literature reports. RESULTS： Thirteen compounds were isolated from ethyl acetate extract part of B. involucrate， identified respectively as pyracanthoside （1）， 5，7，3′ ，5′ -tetrahydroxyflavanone （2）， naringenin （3）， homoeriodictyol （4）， hesperetin （5）， sakuranetin （6）， eriodyctiol （7）， aureusidin-6-O-β-D-glucopyranoside （8）， penicillic acid （9）， dihydropenicillic acid （10）， 2-methyl-3-foroic acid （11）， 5-hydroxymaltol （12） and 5， 7-dyhydroxy chromone （13）. Most of them were dihydroflavones. Compounds 2 to 13 are isolated from Balanophora genus for the first time. CONCLUSIONS： The study enriched the chemical constituents of the Balanophora genus and lays foundation for quality evaluation of B. involucrate.
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Objective To investigate the inhibition of aldose reductase (AR) and antioxidant activity of fractions of ethyl acetate extract from Toona Sinensis seeds. Methods The antioxidant activities of fractions of ethyl acetate extract from Toona Sinensis seeds was evaluated with total antioxidant capacity and DPPH?free radicals scavenging. The inhibition and inhibition types of AR were investigated and the relationship of antioxidant activity and AR inhibitory activity was analyzed. Results Fractions of ethyl acetate extract from Toona Sinensis seeds exhibited antioxidant activity, total antioxidant capacity ordered by Fr.B>Fr.A>Fr.D>Fr.C. The DPPH?free radicals scavenging were ordered by Fr.A>Fr.B>Fr.D>Fr.C. The fractions exhibited AR inhibitory activity and the order was Fr.B>Fr.A>Fr.C>Fr.D. The inhibition mechanism of Fr.B was noncompetitive inhibition and IC50 was 0.401 mg/ml. The AR inhibitory activity of fractions was related to antioxidant activity with correlation index over 0.9. Conclusion This study will provide theoretical basis for exploitation of Toona Sinensis seeds.
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AIM:To investigate the effects of ethyl acetate(EtOAc)extract of Pleione bulbocodioides (Franch.)Rolfe on proliferation and apoptosis of human leukemia K 562 and HL-60 cells and the possible apoptosis path-way.METHODS:Human leukemia cell lines were treated with EtOAc extract of Pleione bulbocodioides at different con-centrations.XTT method was used to evaluate the viability of K 562 cells and HL-60 cells.The cell growth inhibition was calculated by Trypan blue exclusion test.The percentage of apoptotic cells was determined by flow cytometry,and 4,,6-dia-midino-2-phenylindole(DAPI)was used to observe morphological changes of the cells.The cell cycle was observed by pro-pidium iodide(PI)staining.The protein expression of Bcl-2, Bax, cleaved poly(ADP-ribose)polymerase(PARP), cleaved caspase-3,cytochrome C and apoptosis-inducing factor(AIF)wase determined by Western blot.RESULTS:The cell viability and proliferation were inhibited by EtOAc extract of Pleione bulbocodioides with IC50of(42.14 ±2.54)mg/L for HL-60 cells and(51.28 ±3.12)mg/L for K562 cells at 24 h.The results of Annexin V-FITC/PI and DAPI staining showed that EtOAc extract of Pleione bulbocodioides induced cell apoptosis in a dose-dependent manner.The apoptotic rate was increased compared with control group(P<0.05).The G2phase increased with typical cell apoptosis-induced mor-phological changes.The levels of pro-apoptotic proteins Bax,cleaved PARP and cleaved caspase-3 were increased, while Bcl-2 was down-regulated(P<0.05).Cytochrome C and AIF in cytosol,characteristic proteins of intrinsic mitochondrial apoptosis pathway,also increased with the concentration of EtOAc extract of Pleione bulbocodioides increasing(P<0.05). CONCLUSION:EtOAc extract of Pleione bulbocodioides significantly inhibits cell proliferation and induces cell apoptosis in human leukemia cell lines HL-60 and K562 through intrinsic mitochondrial apoptosis pathway.
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OBJECTIVE:To study the inhibitory effect and mechanism of Inula helenium ethyl acetate extract(IHE)on prolif-eration of human pancreatic cancer Capan-2 cells. METHODS:MTT was used to determine the cell proliferation inhibition rate af-ter treated by 0,0.5,1,2,4,8 μg/mL IHE;clone formation test was used to observe the effects of 0,1,2 μg/mL IHE treating for 1 week on cell clone formation;Hoechest 33342 staining was used to observe the changes of nuclear morphology after treated by 0,2,4 μg/mL IHE for 48 h;flow cytometry was used to detect the cell apoptosis rate after treated by 0,4,8,16 μg/mL IHE for 48 h;JC-1 staining was used to observe the changes of intracellular mitochondrial membrane potential after treated by 0,4,8, 16 μg/mL IHE for 24 h;Western blot was used to detect the expressions of mitochondrial apoptosis-related proteins Bcl-2,Bax, Mcl-1,p53 up-regulated modulator of apoptosis (PUMA),and polymerase (PARP) after treated by 0,4,8,16 μg/mL IHE for 48 h. RESULTS:2,4,8 μg/mL IHE had obvious inhibitory effect on cell proliferation,showing concentration-dependent relation-ship,with IC50 of 6.6 μg/mL;1,2 μg/mL IHE can obviously inhibit the clone formation of cells;4 μg/mL IHE can obviously cause cell nuclear condensation;8,16 μg/mL IHE can obviously promote the cell apoptosis,and the cell apoptosis rate reached 45.53% after treated by 16 μg/mL IHE for 48 h;16 μg/mL IHE treating for 24 h can cause the decrease of 82.47% cells'mito-chondrial membrane potential;8 μg/mL IHE can obviously down-regulate the protein expressions of Bcl-2,Mcl-1,PUMA and PARP,and 16 μg/mL IHE can obviously down-regulate the expressions of Mcl-1 and PUMA. CONCLUSIONS:IHE may show its inhibitory effect on proliferation of human pancreatic cancer Capan-2 cells by causing the decrease of mitochondrial mem-brane potential in cells and down-regulating the protein expres-sions of Mcl-1 and PUMA to cause cell apoptosis.
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To investigate the hypoglycemic effects of ethyl acetate extract of Alismatis Rhizoma (AREEA) on type 2 diabetes mellitus,the model of type 2 diabetic rats was induced by high-fat diet feeding for 4 weeks and then intraperitoneal injection of a low dose of streptozotoin (STZ).Rats without the above-mentioned treatment were selected as the normal control group.The diabetic rats were randomly divided into 5 groups:the model control group,low doses (20 mg/kg),medium doses (50 mg/kg),high doses (100 mg/kg) of AREEA groups and positive control-metformin group (100 mg/kg).After four weeks,oral glucose tolerance test (OGTT) and insulin tolerance test (ITT) were performed,respectively.12 hours after the last administration,the levels of serum glucose,glycated hemoglobin(HbA1 c),insulin (Ins),total cholesterol (TC),triglyceride (TG),high density lipoprotein (HDL-C),low density lipoprotein (LDL-C),superoxide dismutase (SOD),malondialdehyde (MDA),glutathione (GSH-Px),tumor necrosis factor-or (TNF-α),interleukin-6 (IL-6),insulin-mediated tyrosine of IRS-1 and Akt phosphorylation in adipose tissue were determined.In addition,the pathological changes of pancreas were examined.After administration for 4 weeks,all doses of AREEA significantly reduced the fasting blood glucose of type 2 diabetic rats (P ＜0.05).In the high doses group of AREEA,the levels of GLU,HbA1c,TC,TG,MDA,TNF-α and IL-6 in serum were decreased significantly,and the levels of SOD and GSH-Px were increased (P ＜ 0.05).These results suggest that AREEA has the therapeutic effect on type 2 diabetes-related symptoms by metabolic regulation of glucose and lipids.
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The ethyl acetate extract of Euphorbia lunulata Bge was separated and researched on anticancer activity about it with in vitro cell experiments,in order to provide a theoretical basis for the exploit and utilization of E.lunulata and more options to develop natural anticancer drugs.Quercetin and Gallic Acid in ethyl acetate were extracted by HPLC,which the ethyl acetate extract of E.lunulata was separated and extracted by solvent system.The MTT was used to measure the inhibition of each component with different concentration from ethyl acetate extract to the 2R-75-30 breast cancer cell.The results showed that Quercetin and Galic Acid of ethyl acetate extract were 19.99％ and 55.04％,respectively.There were 17 component from the ethyl acetate extract,and finally merged into six fractions.Each component (the concentration of 50 μg/ml-150 μg/ml range) from ethyl acetate extract of E.lunulata could inhibit the growth of ZR-75-30 breast cancer cell.And a certain gradient relationship with the drug concentration increased with the inhibition rate enhancement on the 2R-75-30 breast cancer cell's growth.The 2nd fraction showed the highest activity.It was concluded that there were flavonoids and phenolic acids in ethyl acetate extract.Each component from the extract showed inhibition on the growth of breast cancer cell if it was further separated.This study was designed to provide a theoretical basis to exploit the anticancer drugs from E.lunulata.
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This study was aimed to observe the effect of Bai-Zhu Huang-Qi (BZHQ) decoction ethyl acetate extract on NOD like receptor family,pyrin domain-containing 3 (NLRP3) inflammasome in macrophages.The U937 cells were pretreated with phorbol 12-myristate 13-acetate (PMA,10 ng· mL-1) for 48 hours to induce macrophages.Effects on cell viability by different doses of BZHQ decoction ethyl acetate extract (0,3.125,6.25,12.5,25,50,100 μg· mL-1) were observed to select the appropriate concentration.Contents of NLRP3 and caspase-1 in cells were detected by real-time PCR and western blot.The concentration of interleukin-1β (IL-1β) in cell supernatant was detected by enzyme linked immunosorbent assay (ELISA).The cell counting kit-8 (CCK-8) assay showed that when the drug concentration was lower than 25 μg· mL-1,there was no impact on cell viability;when the drug concentration was higher than 50 μg· mL-1,there was inhibition on cell viability (P ＜ 0.05).The concentration of 25 μg· mL-1 was used to conduct the following experiment.Compared to the blank group,the expression of NLRP3 and caspase-l in cells of the LPS group were significantly increased (P ＜ 0.01).The concentration of IL-1β in cell supernatant was also significantly increased (P ＜ 0.01).After treated with BZHQ decoction ethyl acetate extract,levels of NLRP3,caspase-1 and IL-1β were significantly decreased (P ＜ 0.05).It was concluded that BZHQ decoction ethyl acetate extract can inhibit the production of NLRP3 inflammasome in LPS-stimulated macrophages.
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AIM To investigate the effects of ethyl acetate extract from Huangqi Injection (HQIEACE) on leucopenia mice.METHODS An experimental mouse model of leucopenia was induced by cyclophosphamide.NMR based metabolomic profiling technique coupled with multivariate statistical method was used for performing metabolomic analysis.RESULTS HQIEACE could elevate the levels of white blood cell,monocytes,neutrophils and lymphocyte in modeled mice.The levels of ten potential endogenous metabolites (lipid,leucine,3-D-hydroxybutyrate,lactate,alanine,pyruvate,creatine,scyllo-inositol,betaine and glucose) were reversed.CONCLUSION The metabolic pathways related to the pharmacological effects of HQIEACE on leucopenia are probably involved in body energy metabolism,amino acid metabolism,oxidative stress and choline metabolism.
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OBJECTIVE:To study the lifespan effect of ethyl acetate extract from Polygonum multiflora(EPM)on caenorhab-ditis elegans,and to investigate its anti-aging effect. METHODS:EPM low-concentration,medium-concentration and high- concen-tration groups(25,37.5,50 mg/L,calculated by crude drug)and blank group(0 mg/L)were established to investigate the effects of EPM on the lifespan of caenorhabditis elegans. EPM group(37.5 mg/L)and blank group(0 mg/L)were established to perform re-productive test,acute heat stress test and acute oxidative stress test of caenorhabditis elegans. The effects of EPM on reproductive capacity and pressure stress of caenorhabditis elegans were investigated. RESULTS:The lifespan test,average lifespan of cae-norhabditis elegans in EPM low-concentration,medium-concentration and high-concentration groups were prolonged significantly, compared to blank group(P<0.05 or P<0.01),especially in EPM medium-concentration group. In reproductive test,the number of offspring of caenorhabditis elegans in EPM group on the second and third day were increased significantly,compared to blank group (P<0.05). In acute heat stress test and acute oxidative stress test,average survival time of caenorhabditis elegans in EPM group was prolonged significantly(P<0.05). CONCLUSIONS:37.5 mg/L EPM can retard the aging process of caenorhabditis ele-gans and doesn’t damage the reproductive capacity.
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Aim To observe the effects of the ethyl ace-tate extract from Coreopsis tinctoria on glucose and lipid metabolism and liver, kidney function in diabetic rats. Method By high-sugar, high-fat diet combined with intraperitoneal injection of streptozotocin ( streptozoto-cin, STZ ) Type 2 diabetes SD rat model was estab-lished. Model rats were randomly divided into six groups ( control group, model group, three dose groups Coreopsis tinctoria extract:low, middle,high 0. 15 g· kg-1;0. 3 g·kg-1;0. 6 g·kg-1 , positive drug met-formin 0. 16 g · kg-1 group ) . The control group and the model group were given physiological saline and the remaining groups intragastric administration coreofosis tinctoria extrat. Random blood glucose and body weight of rats were measured weekly. After 4 weeks of admin-istration, The rats were killed and rat serum was col-lected to detect serum lipids ( TC/TG/HDL/LDL ) , liver and renal function indicators, serum insulin, and glycated hemoglobin levels. Result Coreopsis tincto-ria ethyl acetate extract effectively reduced the diabetic rats random blood glucose, glycated hemoglobin,serum triglycerides, LDL, total serum protein, serum creati-nine and uric acid levels, and increased serum white protein content in diabetic rats. Conclusion Coreop-sis tinctoria ethyl acetate extract can reduce blood glu-cose and lipid in diabetic SD rats and protect their liver and kidney function.