The result on anatomical study of Inula Helenium / Монголын эм зүй, эм судлал

Introduction@#In recent years, some species of plants that are used in traditional medicine and have high practical value have been successfully introduced in our country. It is necessary to carry out phytochemical studies of local medicinal and useful plants to produce food and biologically active food supplements. The tall coin flower (<i>Inula Helenium</i> L.) has been successfully cultivated in Mongolia, and the lower layer of the plant such as roots and stems are used. However, most of the times the top layer of the plant is not used and thrown away, that is why the plant must be fully utilized. Therefore, the aim of this study is to analyze the top layer of the plant and present the results of anatomical studies.@*Methods@#Remove the top layer of the plant and soak it in glycerin for 24 hours. When making an incision, select a green, undamaged part of the plant and place it in the VCM-202III freezer microtome. Chloral hydrate liquid should be used for micro-preparation. In addition to implementing chloral hydrate solution, use the solutions of various concentrations of sodium alkali (NaOH) (5- 15%). Place the slice in alcian blue stain (using the cell wall staining method) for 3 minutes and wash it with distilled water. Then stain it in a drop of safranin staining (0.5–1.0% aqueous solution) for 1 minute. Rinse it twice with distilled water, add a mixture of glycerin and distilled water, cover it with a glass slide and prepare a temporary slide. Examine the prepared temporary slide with a NOVEL light microscope. Cell images of the plant’s anatomical structures are captured on a computer screen with the help of a digital camera.@*Conclusion@#The anatomy of the leaves, stems and flowers of the tall coin flower (<i>Inula Helenium</i> L.), a plant cultivated in Mongolia, has been analyzed.

In vitro and in vivo antioxidant potentials of the methanolic crude extract from Inula glomerata Oliv. & Hiern (Asteraceae) and Salacia kraussii (Harv) Harv (Celastraceae) / Potenciales antioxidantes in vitro e in vivo del extracto crudo metanólico de Inula glomerata Oliv. & Hiern. (Asteraceae) y Salacia kraussii (Harv) Harv (Celastraceae)

Reactive oxygen species are implicated in multiple pathological conditions including erectile dysfunction. This study evaluated the in vitro and in vivo antioxidant potential of the methanolic extracts of Inula glomerata and Salacia kraussii. The plant materials were pulverized and extracted with methanol. The phytochemical analysis, ability of the crude extracts to scavenge free radicals (ABTS, DPPH, NO.) in vitroas well as the total phenolic and flavonoid contents was investigated. In vivo, antioxidant potentials of the crude extracts (50/250 mg/kg body weight) were determined in an erectile dysfunction rat model. The phytochemical analysis revealed that both plants contain flavonoids, tannins, terpenoids, and alkaloids. The crude extracts at varying degree of efficiency, scavenged ABTS and DPPH radicals. The crude extracts at low concentrations (50 mg/kg b.w) significantly (p<0.05) diminished the level of malondialdehyde, augmented catalase activities and elevated glutathione levels. However, SOD activities were significantly boosted in a dose-dependent manner by the crude extracts. Therefore, I. glomerataand S. kraussiipossess antioxidant properties, hence, can serve as a therapeutic modality in the treatment of oxidative stress-induced erectile dysfunction.

Las especies reactivas de oxígeno están implicadas en múltiples condiciones patológicas, incluyendo la disfunción eréctil. Este estudio evaluó el potencial antioxidante in vitro e in vivo de extractos metanólicos de Inula glomeratay Salacia kraussii. Los materiales vegetales fueron pulverizados y extraídos con metanol. A estos extractos crudos se les llevó a cabo el análisis fitoquímico junto con el contenido total de fenólicos y flavonoides, así como se les investigó la capacidad in vitro para atrapar radicales (ABTS, DPPH, NO.). Los potenciales antioxidantes in vivo de los extractos crudos (50/250 mg/kg de peso corporal) se determinaron en un modelo en ratas con disfunción eréctil. El análisis fitoquímico reveló que ambas plantas contuvieron flavonoides, taninos, terpenoides y alcaloides. Los extractos crudos con un grado variable de eficiencia, atraparon a los radicales ABTS y DPPH. Los extractos crudos a bajas concentraciones (50 mg/kg p.c) significativamente (p<0.05) disminuyeron el nivel de malondialdehído, aumentaron las actividades de catalasa y elevaron los niveles de glutatión. Sin embargo, las actividades de SOD por los extractos crudos fueron significativamente dosis-dependientes. Así, los extractos de I. glomeratay S. kraussii mostraron propiedades antioxidantes, y por lo tanto, podrían servir como una alternativa terapéutica en el tratamiento de disfunción eréctil inducida por estrés oxidativo.

Variations of Chemical Components in Inula japonica by UPLC-MS/MS before and after Honey-frying / 中国药房

OBJECTIVE：To compare the c hemical components differences of Inula japonica before and after honey-frying. METHODS：UPLC-MS/MS method was adopted. The determination was performed on Waters ACQUITY UPLC BEH C 18 column with mobile phase consisted of 0.1％ formic acid-acetonitrile （gradient elution ）at the flow rate of 0.3 mL/min. The column temperature was set at 30 ℃，and sample size was 5 µL. The electrospray ion source was scanned by positive ion mode. The first order mass spectrometry scanning range was m/z 70-1 050，the second order mass spectrometry scanning range was m/z 50-1 050， and the normalized collision energy was 40，60 eV ；mass spectrum type was the peak figure ，the flow rate of sheath gas was 35 arb，the auxiliary airflow speed was 10 arb，the spray voltage was 3.80 kV，the S-lens voltage was 50 V，the heating temperature was 350 ℃，and the capillary temperature was 350 ℃. The components were identified by Qual Browser 4.1.39.1 software， referring to the online high-resolution database mzCloud and local database OTCML of high-resolution mass spectrometry of TCM ， and combined with relevant literature. The principal component analysis （PCA）and orthogonal partial least squared-discriminant analysis（OPLS-DA）of I. japonica before and after honey-fried were performed by using SIMCA 14.1 statistical software ，and variable importance projection （VIP）value greater than 1 was used as the standard to screen the differential components before and after honey-frying. RESULTS ：A total of 29 common chemical components were identified from I. japonica and honey-fried I. japonica，including 5 phenolic acids as 1-caffeoylquinic acid ，chlorogenic acid and 3，5-dicaffeoylquinic acid ，12 flavonoids as quercetin，luteolin and evamectin ，as well as 12 sesquiterpene lactones as 1-O-acetylinula diester ，inula bicolor lactone B and 1-O-acetyl-6-O-isobutyryl inulin. The results of PCA showed that I. japonica and honey-fried I. japonica were located on both sides of the score diagram respectively. The results of OPLS-DA showed that the VIP values of 7 components were greater than 1，which were peak 19（britanin），peak 6（quercetagitrin），peak 1（1-caffeoylquinic acid ），peak 21（vitexicarpin），peak 20（tomentosin）， peak 13（spinacetin）and peak 3（daphnetin）. CONCLUSIONS ：After honey-fried ，the content of chemical components of I. japonica changed and decreased to a certain extent. Britanin ，quercetagitrin，1-caffeoylquinic acid ，tomentosin，vitexicarpin， spinacetin and daphnetin may be the differential components of I. japonica and honey-fried I. japonica .

Simultaneous Determination of Seven Components in Blossoms of Inula nervosa by UHPLC / 中国实验方剂学杂志

Objective::To establish an ultra-high-performance liquid chromatography (UHPLC) method for simultaneous determination of seven components(chlorogenic acid, 3, 5-dicaffeoylquinic acid, 1, 5-dicaffeoylquinic acid, kaempferol-3-O-rutinoside, quercetin, kaempferol and thymol) in blossoms of Inula nervosa, and provide references for its quality control. Method::The separation was performed on an Agilent Poroshell 120 C18 column (3.0 mm×100 mm, 2.7 μm) with 0.1% formic acid(A) and methyl (B)as mobile phase for gradient elution(0-4 min, 2%B; 4-6 min, 2%-5%B; 6-10 min, 5%-10%B; 10-20 min, 10%-20%B; 20-30 min, 20%-27%B; 30-37 min, 27%-25%B, 37-45 min, 25%-32%B; 45-68 min, 32%-58%B; 68-75 min, 58%-25%B; 75-82 min, 25%-2%B; 82-90 min, 2%B). The flow rate was 0.3 mL·min-1 and the detection wavelength was 254 nm. Result::There was a good linear relationship between the concentration and peak area of all the seven components in the investigated concentration range (r>0.999). The average recoveries ranged from 97.80% to 101.28% with RSD≤3.0%. Cluster analysis of SPSS software and principal component analysis of SIMCA software can be used to intuitively classify samples from four different origins. Conclusion::The established method is simple and fast with high precision, which can be used to compare the differences of blossoms of Inula nervosa from different origins and efficiently control its quality.

Effect and mechanism of Inula britannica flower total flavonoids on the expression of lncRNA TERRA in the senescent skin fibroblasts / 解放军医学杂志

Objective: To investigate the effect of Inula britannica flower total flavonoids (IBFTF) on the expression of lncRNA TERRA in the senile skin fibroblasts and its mechanism. Methods: High performance liquid chromatography (HPLC) was used to analyze the composition and content of IBFTF. The human skin fibroblasts cell line was divided into control group, IBFTF group (cells were treated with IBFTF alone), cell senescence model group [cell senescence model was induced by D-galactose (D-gal)] and cell senescence model + IBFTF group (cell senescence model was treated with IBFTF). The cell viability was detected by CCK-8. The senescence-associated β-galactosidase (SA-β-Gal) staining was used to detect the positive rate of human skin fibroblasts cells, and the contents of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and malondialdehyde (MDA) were detected by enzyme microplate reader. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of lncRNA TERRA, hTERT mRNA and the relative length of telomere in the cells. The protein expression of agingrelated proteins such as p53, p16 and human telomerase reverse transcriptase (hTERT) were detected by Western blotting. Results: The contents of quercetin, isorhamnetin, kaempferol and rutin in IBFTF were 6.447, 2.044, 1.272, 0.781 mg/g, respectively. CCK-8 and SA-β-Gal staining showed that IBFTF increased the viability of cells in senescence model group, and reduced the proportion of SA-β-Gal positive cells (P<0.05). Enzyme labeling results showed that IBFTF increased the activity of SOD and GSH-Px, and reduced the MDA level of senescent cells (P<0.05). qRT-PCR showed that IBFTF decreased the expression of lncRNA TERRA, increased the expression of hTERT mRNA and prolonged the telomere length in senescent cells (P<0.05). Western blotting results showed that, compared with cell senescence model group, the protein expressions of p53, p16 decreased, and the hTERT increased in cell senescence model + IBFTF group (P<0.05). Conclusions: IBFTF can decrease the expression of lncRNA TERRA in D-galinduced senescent human skin fibroblasts cells, and its mechanism may be related to the recovery of telomerase activity and the prolongation of telomere length, so as to achieve the effect of delaying senescence.

Comparative analysis of three complete chloroplast genomes of Inula genus with phylogenetic analysis of 49 plants from Carduoideae / 药学学报

italic>Inula japonica, Inula hupehensis and Inula linaariifolia are all medicinal plants of Inula L. in the Compositae family, and Inula hupehensis is endemic to China. In order to compare their genomic sequence differences and provide scientific basis for their germplasm conservation and development, we obtained and analyzed the complete chloroplast genomes of these three species. Total DNAs were extracted from fresh leaves and subjected to next-generation DNA sequencing. NOVOPlasty was used to assemble the chloroplast genomes from the sequence reads. CPGAVAS2 was used to annotate the genes and repeats in each genome. Lastly, phylogenomics analysis was conducted using RAxML. The results showed that the total length of the chloroplast genome of Inula japonica, Inula hupehensis and Inula linaariifolia is 150 754, 150 909, and 150 812 bp respectively, each consisting of a large single copy region, a small single copy region and a pair of inverted repeat regions. In addition, the G/C content of all three chloroplast genomes was approximately 37.7% and each encoded 111 unique genes, including 79 protein-coding, 28 tRNA and 4 rRNA genes. Meanwhile, 32, 33, 34 simple repeat sequences, 18, 22, 18 tandem repeat sequences and 33, 37, 38 scattered repeat sequences were identified in three species. Phylogenomic analysis showed that all three species of Inula L. and Pluchea indica were clustered together, with the relationship between Inuleae and Senecioneae closer, suggesting that Inuleae may have originated from the Senecioneae, not the Cardueae. The data in this study not only enriches the chloroplast genome database of Inula L., but also lays the foundation for the future studies of species identification, phylogenetic relationships, evolution history and genetic diversity of Inula species.

Determination of britanin content in different parts of Inula linariifolia Turcz by HPLC / 药学实践杂志

Obejctive To determine and compare britanin content in dried aerial parts and capitulum of Inula linariifolia Turcz from 7 different origins by HPLC. Methods Analysis was performed on Agilent Zorbax SB C18 (250 mm×4.6 mm, 5 μm). Acetonitrile and water were used as mobile phase for gradient elution at 1.2 ml/min. Column temperature was 30 ℃ and the detection wavelength at 212 nm. Results The results meet the requirements of the method validation in 2015 edition of Chinese Pharmacopeia. The average britanin content in dried aerial parts of Inula linariifolia Turcz is 0.125% vs 0.732% in capitulum, which is significantly different. Conclusion The established method is simple and convenient. It can be used for quality control of Inula linariifolia Turcz.

Effect of Inula cappa on CYP450 Enzyme in Rats by Cocktail Probe Drugs / 中国实验方剂学杂志

Objective:To study on the effect of Inula cappa extract on the activities of six cytochrome P450(CYP450) enzymes in rats by Cocktail probe method. Method:Rats in the I. cappa high and low dose groups were given oral administration of active fractions of I. cappa at a dose of 8.127,2.709 g·kg-1·d-1 of the material for 7,14 d,repectively.Probe drugs(caffeine,midazolam,tolbutamide,omeprazole,metoprolol,chlorzoxazone) were simultaneously injected to rats after administration.Plasma was collected at different time after the administration of probe drugs.The plasma concentrations of these six probes were measured by UPLC-MS and their corresponding pharmacokinetic parameters were calculated with DAS 2.0. Result:Compared with the control group,only the apparent volume of distribution(V) of midazolam was increased;area under the curve(AUC0-t and AUC0-∞)and half-life period(T1/2) of caffeine,midazolam,tolbutamide and omeprazole were increased and the clearance rate(CL) of them was decreased in rats of I. cappa groups.But there were no differences in pharmacokinetic parameters of chlorzoxazone and metoprolol. Conclusion:I. cappa can reduce the enzymatic activities of CYP3A,CYP1A2,CYP2C9 and CYP2C19 in rats at different degree,among which CYP3A is the strongest.

Determination of plasma protein binding rates of nine compounds of Inula cappa extraction based on method of equilibrium dialysis / 中国中药杂志

To determine the plasma protein binding rate of the nine compounds in Inula cappa extraction by the method of equilibrium dialysis. The proteins in plasma samples were precipitated by methanol, and the ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) was developed for determination of the concentrations of the nine active compounds, namely chlorogenic acid, scopolin, neochlorogenic acid, cryptochlorogenic acid, 1,3-O-dicaffeoylquinic acid, galuteolin, 3,4-O-dicaffeoylquinic acid, 3,5-O-dicaffeoylquinic acid, 4,5-O-dicaffeoylquinic acid, with the internal standard of puerarin. We found that all components have a good linearity(r≥0.999), and accuracy, precision, extraction recovery and stability conformed to the requirements of determination, without endogenous compounds disturbing within the range of optimum concentration. This suggested that the method was stable and reliable, and could be used for the determination of the plasma protein binding rates of the nine active compounds in rat and human plasma of I. cappa. The plasma protein binding rates of the nine active compounds in rat and human plasma respectively were(41.07±0.046)%-(94.95±0.008)%, and(37.66±0.043)%-(97.46±0.013)%. According to the results, there were differences in the plasma protein binding rates of the nine compounds in I. cappa extraction between rat and human.

Anti-inflammatory Activity of Standardized Fraction from Inula helenium L. via Suppression of NF-κB Pathway in RAW 264.7 Cells

Inula helenium L. is rich source of eudesmane-type sesquiterpene lactones, mainly alantolactone and isoalantolactone, which have the various pharmacological functions. In this study, we examined the inhibitory effects of nitric oxide (NO) production of hexane, methylene chloride, ethyl acetate, butanol, and water fractions from I. helenium and investigated the anti-inflammatory effect of hexane fraction of I. helenium (HFIH) in LPS-induced RAW 264.7 cells. Quantification of alantolactone and isoalantolactone from HFIH was carried out for the standardization by multiple reaction monitoring using triple quadrupole mass spectrometer. HFIH significantly inhibited inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) protein as well as their downstream products NO and prostaglandin E₂ (PGE₂) in LPS-stimulated RAW 264.7 cells. Moreover, HFIH suppressed NF-κB transcriptional activity by decreasing the translocation of p65 to the nucleus. The in vivo study further confirmed that HFIH attenuated the paw edema induced by carrageenan in an acute inflammation model. These findings suggest that HFIH may be useful as a promising phytomedicine for inflammatory-associated diseases.

Investigation of the Excretion of Seven Constituents of Inula cappaort in Rat Urine by UPLC-MS/MS / 中国药学杂志

OBJECTIVE: To establish an UPLC-MS/MS method for the analysis of seven compounds of Inula cappaort in rat urine to study their excretion. METHODS: The urine samples in 0-2, 2-6, 6-12, 12-24, and 24-36 h were collected. Acquity UPLC BEH C18 column (2.1 mm×50 mm, 1.7 μm) was used and the column temperature was set at 45 ℃, the mobile phase was 0.1% formic acid acetonitrile -0.1% formic acid aqueous solution in a gradient elution mode and flow rate was 0.25 mL•min-1. The detection was carried out by a triple quadrupole linear ion trap mass spectrometer in positive and negative ion mode with an electrospray source. Multiple reactions monitoring (MRM) mode was employed. RESULTS: The calibration curves showed good linearity, with correlation coefficients of greater than 0.991 0 for all of the analytes within the concentration ranges. The intra-day and inter-day precisions (RSD) were all less than 15%. The extraction recoveries of the seven components were more than 84.41%, without obvious matrix effect, which met the requirements for analysis. CONCLUSION: The established method is simple, rapid, and sensitive. It can be applied in the excretion study of the seven components of Inula cappaort extract in rat urine. The urine excretion test showed that the prototype excretion rates are low in rats, and the cumulative excretion rates are all less than 5%.

Effect of nitrogen,phosphorus and potassium formula fertilizer on growth and chemical composition content of Inula japonica / 中国中药杂志

Inula japonica was used as the research object, "3414" fertilization experiment were conducted to study the effects of nitrogen,phosphorus and potassium formula fertilizer on the growth and chemical composition content of I. japonica. The characteristics of fertilizer requirement were preliminarily revealed and the study provided fertilization guidance for artificial cultivation of I. japonica. The results showed that different nitrogen,phosphorus and potassium formula fertilizers had significant effects on plant morphology,physiological and biochemical indexes,dry matter accumulation and chemical composition content. The growth indexes and chemical components of I. japonica showed an upward trend with the increase of fertilization amount,especially the nitrogen fertilizer was the most significant. The indicators were analyzed by membership function. After comprehensive evaluation,the optimal nitrogen,phosphorus and potassium formula fertilization level was N3 P2 K2,namely high level nitrogen fertilizer,medium level phosphorus fertilizer and potassium fertilizer. I. japonica is a high fertilizer demand plant,and the rational fertilization scheme is " applying nitrogen fertilizer again and applying phosphorus and potassium fertilizer properly".

Inula britannica flower total flavonoids inhibiting expression of RAGE in aging L929 cells induced by advanced glycation end products / 中草药

Objective To investigate the protective effect of Inula britannica flower total flavonoids (IBFTF) on aging L929 cells induced by advanced glycation end products (AGEs), and to explore the mechanism of the expression of receptor advanced glycation end products (RAGE). Methods The senescence L929 cells model was established by using 0.100 g/L AGEs for 48 h culture in vitro, and then different concentration of IBFTF (0.1, 0.2, and 0.4 g/L) was given to the treatment group for 6 h culture, and 0.1 g/L aminoguanidine hemisulphate (AG) was given to the positive group for another 6 h culture, and while blank group was cultured with common culture medium. Senescence aging index of β-galactosidase staining cell numbers and cell cycle analysis in L929 cells were measured. The levels of reactive oxygen species associated with oxidative stress in the cells, and aging indicators such as superoxide dismutase (SOD), malondialdehyde (MDA) were also examined. The fluorescent intensity of RAGE was detected by immunofluorescence. The expression of RAGE protein and mRNA was detected by Western blotting and qPCR, respectively. Results IBFTF significantly inhibited AGEs-induced L929 cells senescence and decreased RAGE protein and mRNA expression. Different concentrations of IBFTF could significantly increase the SOD activity and reduce the MDA content and eliminate the reactive oxygen species in aging L929 cells in a dose-dependent manner. Conclusion Protective effect of IBFTF on aging L929 cells induced by AGEs may be related to its effect on the surpressing the expression of RAGE.

Chemical constituents from Inula helenium / 中草药

Objective To study the major chemical components from Inula helenium. Methods The compounds were separated and purifid by using a variety of chromatographic techniques including silica gel column chromatography, thin layer chromatography, and high performance liquid chromatography, and the structures of the compounds were verified by nuclear magnetic spectroscopy and literature data. Results A total of 22 compounds were separated from petroleum ether extract of I. helenium and identified separately as alantolactone (1), isoalantolactone (2), 4,4-dimethylsterols (3), 11αH,13-dihydroisoalantolactone (4), 11αH,13-dihydroalantolactone (5), 4(15)-epoxy-isoalantolactone (6), 5α,6α-epoxyalantolactone (7), alloalantolactone (8), isoalloalantolactone (9), friedelin (10), friedelinol (11), erythrodiol (12), β-sitosterol-glucopyranoside (13), lupeol acetate (14), lupeone (15), lupeol (16), δ-amyrin (17), lupeol palitate (18), 5,7,4'-trihydroxy-3',5'-dimethoxy flavane (19), (+)-syringaresinol (20), 3,5,3'-trihydroxy-6,7,4'-trimethoxy flavone (21), and 3,5,6,7,3'-hydroxy-4'-methoxy dihydroflavones (22). Conclusion Compounds 21 and 22 are isolated from this genus for the first time; Compounds 10-15, 17, and 18 are separated from the I. helenium for the first time. After antibacterial test, compounds 1, 2, 4, 5, 7-9 have strong inhibitory effects on Escherichia coli, Staphylococcus aureus, and Bacillus subtilis.

Absorption of Inula cappa extract based on everted intestinal sac method / 中国中药杂志

To investigate the absorptive characteristics of Inula cappa extract based on the rat everted intestinal sac method . Nine representative ingredients in I. cappa extract were selected as the study objects. An UPLC-MS/MS method was established to determine and detect their cumulative absorption amount for expounding the absorptive characteristics of ingredients in different intestinal sections. According to the results， the transport mechanism of 8 compounds showed passive diffusion by the reverted gut sac method. And scopolin was actively transported in the intestine. The best absorption site of chlorogenic acid was duodenum. The best absorption site of cryptochlorogenic acid， 1，3--dicaffeoylquinic acid， luteolin-7-glucoside and 3，4--dicaffeoylquinic acid were jejunum. The best absorption site of neochlorogenic acid， scopolin， 4，5--dicaffeoylquinic acid and 3，5--dicaffeoylquinic acid was ileum. The absorption of all the compounds was affected by pH and bile. All of the nine ingredients in I. cappa extract could be absorbed in intestines， but with differences in the absorption rate， the best absorptive site and mechanism， indicating that the intestinal absorption of I. cappa extract was selective.

Analysis of components in plasma from effective fractions of Inula cappa based on UHPLC/Q-TOF/MS / 中国药理学通报

Aim To analyze the main components of effective fractions of Inula cappa in rat plasma with UHPLC/Q-TOF/MS technology and serum pharmacochemistry theory.Methods After gavage administration with Inula cappa extracts,blood was collected from abdominal aorta,and the protein in plasma sample was precipitated by methanol.Eclipse Plus C18 RRHD (2.1 mm × 100 mm,1.8 μm) was used,with 0.1％ formic acid agueous solution (A)-0.1％ formic acid and acetonitrile (B) as the mobile phase for gradient elution,and the flow rate was 0.3 mL ·min-1 Detected at negative ion mode,and with the help of Metabolite Tools software from Bruker Corporation,components in plasma were defined.Results A total of 16 compounds were identified,including 9 prototypes and 7 metabolites.Main metabolites were isomerization,methylation,glucuronidation and recombination of caffeoylquinic acid.Conclusions Therefore,our study has comprehensively expounded Inula cappa extracts' constituents migrating to rat plasma,and provided a reference for further studies for in vivo metabolic process and effective material base of Inula cappa.

Study on the Inhibitory Effect and Mechanism of Inula helenium Ethyl Acetate Extract on Proliferation of Human Pancreatic Cancer Capan-2 Cells / 中国药房

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.

Chemical Constituents of Inula cappa / 中国药学杂志

OBJECTIVE: To study the chemical constituents of Inula cappa. METHODS: Chromatographic techniques were employed for isolation and purification of the constituents and their structures were determined by spectral analysis and chemical evidence. RESULTS: Seventeen compounds were obtained and identified as friedelin(1), epifriedelanol(2), α-amyrin(3), β-amyrin(4), benzyl 2-O-β-D-glucopyranosy-2, 6-dihydroxybenzoate(5), scopoletin(6), luteolin-7-O-β-D-glucuronide ethyl ester(7), benzyl alcohol glucoside(8), ophiopogonoside A(9), apigenin-7-O-β-D-glucoside(10), luteolin-7-O-β-D-rutinoside(11), hydnocarpin-D(12), luteolin(13), luteolin-7-O-β-D-glucoside (14), luteolin-4'-O-β-D-glucoside (15), quercetin-3-O-β-D-glucoside (16), and juglans cerebroside A(17). CONCLUSION: Compounds 5, 7, 9, 11, 12, and 17 are for the first time obtained from the genus Inula and compounds 8, 10, 14, and 16 are isolated from Inula cappa for the first time.

Metabolism of Inula cappa extract by rat intestinal bacteria in vitro / 中国中药杂志

To investigate the metabolism of major components in Inula cappa by rat intestinal bacteria in vitro. I. cappa extract was incubated for 24 h with rat intestinal bacteria under anaerobic environment. After the samples were precipitated by n-butanol, UPLC-Q-TOF-MS/MS was applied for the qualitative analysis of the metabolites, combined with data software such as Metabolite Tools, Data Analysis and so on. The potential metabolites in rat intestinal bacteria were analyzed by comparing the total ion current of the test samples and blank samples and analyzing the quasi-molecular ion and fragment ion of all chromatograms. The results injected that fourteen metabolites were detected in rat intestinal flora. Various types of metabolic reactions happen to caffeoylquinic acid in intestinal flora, including isomerization, hydrolyzation, there were also methylation, hydrogenation and acetylation of caffeic acid. At the same time, a methylate of dicaffeoylquinic acid was also detected. Presumably, caffeoylquinic acids were gradually transformed into more hydrophobic metabolites with smaller molecular mass, which were better absorbed by the intestinal tract.

Quantitative determination of seven constituents and fingerprint research on Inula cappa by UPLC / 中草药

Objective: This study is aimed at developing an UPLC method for simultaneous determination of 1, 3-O-dicaffeoylquinic acid, 1,5-O-dicaffeoylquinic acid, 3,5-O-dicaffeoylquinic acid, 4,5-O-dicaffeoylquinic acid, 3-O-caffeoylquinic acid, 4-O-caffeoylquinic acid, and 5-O-caffeoylquinic acid and fingerprint analysis of Inula cappa. Methods: WATERS ACQUITY UPLC BEH C18 column with gradient elution of 0.1% acetic acid-acetonitrile at a flow rate of 0.4 mL/min was employed for analysis of 20 batches of samples from three provinces or autonomous region. The detected wavelength was set at 329 nm. The column temperature was maintained at 30℃ and the sample solutions were maintained at 4℃ before analysis. Results: Twenty peaks were selected as the common peaks in fingerprint and the similarity of samples was above 0.900 except S18. The variance analysis, hierarchical clustering analysis, and principle component analysis were employed for the quality evaluation based on the contents of seven components and fingerprint similarity. The results indicated that the contents of 1,3- and 1,5-O-dicaffeoylquinic acid were significantly different among three provinces or autonomous region. The clustering analysis results showed that 20 batches of samples were divided into two classes and the quality of samples from Guangxi provinves were more stable than those from Guizhou and Yunnan. Conclusion: The established method could be rapid and accurate to evaluate the quality of I. cappa.