Optimization of Extraction Conditions of Polyphenols and Flavonoids from Olax subscorpioidea leaves, a Medicinal Plant Traditionally Used in Korhogo (Cote d’ivoire)

Secondary metabolites, including polyphenols and flavonoids, play a key role in the expression of the active principles of several medicinal plants from Côte d'Ivoire, such as Olax subscorpioidea Oliv. This study was initiated to help improve conditions for extracting polyphenols and flavonoids from Olax subscorpioidea Oliv leaves harvested in Korhogo. Experimental design methodology was used. The effect of four independent variables on polyphenol and flavonoid content was evaluated: extraction time (12h-24h), evaporation temperature (45°C - 55°C), plant material-solvent ratio (1/20 g/mL - 1/5 g/mL) and ethanol proportion (20% - 100%). The Central Composite Design (CCD) was used to optimize responses after factor screening using the Plackett and Burman design. Optimum extraction conditions were obtained with an ethanolic proportion of 88.29% and 17g of Olax subscorpioidea Oliv leaves dissolved in 100 mL ethanol. The experimental values (96.31 ± 5.29 mg Eq AG/g for polyphenols and 62.44 ± 4.58 mg Eq Q/g) were close to those expected (94.51 mg Eq AG/g for polyphenols and 58.22 mg Eq Q/g for flavonoids), confirming the relevance of the mathematical model used and the success of the response surface methodology in optimizing the extraction conditions of the system studied. These results show that by taking extraction conditions into account in this preliminary study, we can optimize the quantities of secondary metabolites of therapeutic interest.

Lok?t (Eriobotrya japonica; family: Rosaceae): Nutritional and Medicinal Properties, in the Light of Unani Medicine and Scientific Studies

Eriobotrya japonica Lind. (Family: Rosaceae) named as loquat, is a subtropical fruit, which is well known medicinal plant cultivated in Japan and China. Various parts, like leaves, peels and fruits have been shown to possess various useful health benefits. In Unani medicine, it is vastly utilized in many illnesses, like fevers, nausea, de-arranged sanguinous humour (diseases due to morbidity of blood), indigestion, liver diseases, vomiting, dysentery, wounds, inflammations etc. Loquat plant contains many active constituents, such as glycosides, flavonoids, polyphenolic compounds, tannin etc. and nutritional and mineral compounds like, carotenoids, vitamins, starch, amino acids, sugar and others. According to various pharmacological studies it is found that the plant has many biological effects like antitussive, anti-melanogenic, anti-diabetic, anti-inflammatory, antimicrobial, kidney protective, hepatoprotective and hypolipidemic activity. This review aims to shed light on the therapeutic applications of loquat based on both traditional Unani literature and scientific studies conducted on different parts of the plant.

Hepatoprotective effect of flavonoid rich fraction of Sesbania grandiflora: Results of In vivo, in vitro, and molecular docking studies

Background: Phytochemicals and their derivatives are promising target drugs for various ailments and have served as therapeutic agents for several decades. Using in vivo and in vitro models and molecular docking, this study investigated the pharmacological potential of a flavonoid-rich fraction of the ethanolic extract of Sesbania grandiflora (SG). Objectives: This research aimed to determine whether flavonoid-rich whole-plant extracts of SGs have any cytoprotective or in vivo hepatoprotective effects. Additionally, the study was intended to elucidate the molecular connections between the discovered flavonoid flavonols and PPAR? target proteins linked to liver problems, for which an in silico molecular docking investigation was performed. Materials and methods: To separate the flavonoid components, the entire Sesbania grandiflora plant was first extracted using ethanol as a solvent by soxhlet extraction. The resulting ethanolic extract was then fractionated. The cytoprotective and hepatoprotective properties were evaluated via in vitro and in vivo experiments. SGOT, SGPT, triglyceride, bilirubin, and total protein levels were used to evaluate hepatotoxicity in animal models. In vitro studies on Hepatocellular Carcinoma G2 (HepG2) cell lines have examined their cytotoxic effects and antioxidant activity. The most promising flavonoid-flavanol compounds were identified by conducting molecular docking studies against PPAR? target protein (PDB ID: 3VI8) using MOE software. Results: In vivo, the serum levels of SGOT, SGPT, total triglyceride and total bilirubin were measured in exper- imental animals treated with the flavonoid-rich ethanolic extract of SG. Significant reductions in the levels of these hepatic injury markers were observed, indicating the hepatoprotective potential of the extract. Elevated levels of liver biomarkers in the untreated group indicated liver injury or dysfunction. The treated groups showed significant restoration of these biomarkers, suggesting the hepatoprotective potential of SG. The IC50 value for the total flavonoid content of SG was 190.28 ?g/ml, indicating its safety in inhibiting HepG2 cell growth. Flavonoid treatment decreased cell viability but did not affect antioxidant parameters in hepatocytes. In addi- tion, SG restored the damaged hepatocyte architecture. Molecular docking studies revealed the binding affinities of flavonoids for PPAR?. These findings suggest that a promising lead candidate for the development of thera- peutic medicines against anti-TB drug-induced hepatotoxicity has been identified. Conclusion: Our findings demonstrate the hepatoprotective potential of the flavonoid-rich fraction of Sesbania grandiflora both in vivo and in vitro. This study provides valuable insights into its mechanism of action, high- lighting its promising therapeutic application in the management of liver disorders. This study highlights the hepatoprotective and cytoprotective potential of the total flavonoid-rich fraction of SG.

Phytochemicals and colorectal cancer: about polyphenols / Fitoquímicos y cáncer colorrectal: acerca de los polifenoles

Dietary consumption of polyphenols, found in fruits and vegetables, has been associated with a potentially protective role in colorectal cancer (CRC). To establish the state of knowledge regarding advances in polyphenols, CCR and action mechanisms a systematic review and an analysis of information available until 2021 were made. Results indicate that only some polyphenols have in vitro, preclinical and clinical studies. These studies showed that polyphenols will inhibit human CRC cell invasion, migration, metastasis formation, tumor growth. Action mechanisms involve signaling pathways that modulate genes, proteins, markers or cell death inductors, like the AMPK pathway, caspases, Bcl-2, p-Akt, and NF-kB, lysosomal and mitochondrial dysfunction, cellular cycle arrest, among the best known and implied in CRC. Overall, in vitro, preclinical and clinical data on phytochemicals against CRC are still not sufficient and therefore the preventive or therapeutic impacts of dietary phytochemicals on CRC development deserve further research.

El consumo dietético de polifenoles, que se encuentran en frutas y verduras, se ha asociado con un papel potencialmente protector contra el cáncer colorrectal (CCR). Para establecer el estado del conocimiento respecto a los avances en polifenoles, CCR y mecanismos de acción, se realizó una revisión sistemática y un análisis de la información disponible hasta el año 2021. Los resultados indican que sólo algunos polifenoles cuentan con estudios in vitro, preclínicos y clínicos. Estos estudios demostraron que los polifenoles inhiben la invasión, migración, formación de metástasis y crecimiento de tumores de células de CCR humanas. Los mecanismos de acción involucran vías de señalización que modulan genes, proteínas, marcadores o inductores de muerte celular, como la vía AMPK, caspasas, Bcl-2, p-Akt y NF-kB, disfunción lisosomal y mitocondrial, parada del ciclo celular, entre los más conocidos e implicados en el CCR. En general, los datos in vitro, preclínicos y clínicos sobre fitoquímicos contra el CCR aún no son suficientes y, por lo tanto, los impactos preventivos o terapéuticos de los fitoquímicos dietéticos en el desarrollo del CCR requieren más investigación.

Non-volatile leaves compounds LC-MS profile of the two Cuban Piper aduncum L. subspecies / Perfil de CL-SM de compuestos no volátiles de las hojas de las dos subspecies cubanas de Piper aduncum L

The medicinal plant Piper aduncum was segregated in Cuba as P. aduncum subsp. ossanumand P. aduncum subsp. aduncum, the latter has not been phytochemically investigated in Cuba. This study aims to compare profiles of non-volatile compounds of the aqueous and ethanolic extracts of Cuban Piper aduncum subspecies leaves. From both subspecies were prepared a decoction and, an ethanolic extract that yield two fractions. Analysis was performed by UPLC-ESI/QTOF/MS. Standard reference compounds were assessed by comparison with MS data of the plants extracts. Also, tentatively assigned compounds were proposed by matching m/z and fragmentation patterns with P. aduncum published compounds. Main compounds in decoction and aqueous ethanolic fraction of both subspecies were characterized as flavon-C-glycosides. Isoorientin was not previously reported in P. aduncum. Oleaginous ethanolic extracts differs among subspecies and peaks are coincident with benzoic acid derivate compounds. Results can serve for quality control of P. aduncumin Cuba

La planta medicinal Piper aduncum, fue segregada en Cuba como P. aduncum subsp. ossanum y P. aduncum subsp. aduncum, la última carece de estudios fitoquímicos en Cuba. El objetivo fue comparar los perfiles de los compuestos no volátiles de extractos acuosos y etanólicos de hojas de las subespecies cubanas de P. aduncum. De ambas subespecies se prepararon decocciones y extractos etanólicos que rindieron dos fracciones, que fueron analizadas mediante UPLC-ESI/QTOF/MS. Los MS obtenidos se evaluaron respecto a compuestos de referencia y tentativamente se asignaron compuestos comparando con los reportados en P. aduncum. Los perfiles de LC-MS de decocciones y fracciones acuosas fueron similares en ambas subespecies, los compuestos mayoritarios fueron C-glucósidos de flavonas. La isorientina no se ha reportado antes en P. aduncum. Las fracciones oleaginosas difieren entre las subespecies, sus picos coinciden con derivados del ácido benzoico. Estos resultados pueden servir para el control de calidad de P. aduncum en Cuba.

Chemical Composition and Oxidative Properties of Moringa Peregrina Dried-Ground Parts Extracted with Different Solvents

Background: Moringa species contain extremely high level of antioxidant and low levels of anti-nutritional compounds distributed between different parts of the plant. Objective: The present study examined the chemical composition, total energy, and oxidative properties of several solvent extracts of Moringa peregrina (MP) seeds, bark, and leaves dried-ground. Methods: The chemical composition, total energy, and mineral content of the MP parts were all determined. The total phenolics, total flavonoids, and antioxidant activity of various solvent extracts were also measured. Results: The chemical composition differed between parts, with seeds having considerably (p ? 0.05) high levels of protein (19.4%), fat (32.23%), and ash (2.90%). The bark was significantly (p ? 0.05) higher in carbohydrates (76.1%) than the other parts. The leaves had significantly (p ? 0.05) higher levels of Na, Mg, K, Mn, Fe, Zn, and Se. However, the seeds are rich in P and Cu, whereas the bark is rich in calcium. The solvents' efficacy in extracting phenolics and flavonoids from the three parts varied substantially, with aqueous methanol ranking first and the leaves having larger amounts than the other parts. The antioxidant activity was examined using the DPPH, ABTS, and FRAP tests. Leaves had stronger antioxidant activity than other parts. Furthermore, antioxidant activity was strongly associated with total phenolics and flavonoids. Conclusion: In general, extracts made with aqueous organic solvents produced more total phenolics and flavonoids and had higher antioxidant activity, with leaves demonstrating superior antioxidant properties compared to other parts.

Optimization of microwave-assisted extraction of total flavonoid content from red betel leaf (Piper crocatum Ruiz and Pav) and its correlation with antioxidant and antibacterial activities using response surface methodology

The optimization of microwave-assisted extraction (MAE) was conducted using response surface methods to improve the extraction of flavonoids from Piper crocatum Ruiz and Pav leaf. The optimization process included a Box–Behnken experimental design (BBD), which involved three variables at three levels. The present study aimed to evaluate the impact of varying ethanol concentrations (50%, 75%, and 100%), microwave power levels (180, 300, and 450 W), and extraction durations (3, 8.5, and 14 minutes) on the respective responses. The experimental data was subjected to fitting using a second-order polynomial model. Subsequently, an analysis of variance (ANOVA) and multiple regression analysis were employed to assess the adequacy of the model and determine the ideal settings. Taking into account the highest concentration of extracted total flavonoids, as well as the antioxidant and antibacterial properties. The experimental results indicate that the optimum conditions for all the reactions under investigation were an ethanol concentration of 78.48%, a microwave power of 327.96 W, and an extraction duration of 8.60 minutes. Under the ideal conditions, the anticipated outcomes of the sample indicate a total flavonoid content (TFC) of 229.647 mg QE/g dry weight (DW), a 1,1-diphenyl-2-picryhydrazyl (DPPH) scavenging activity of 73.915%, and an inhibition zone measuring 18.621 mm. The implementation of a concurrent MAE methodology for the isolation of total flavonoids, as well as the evaluation of antioxidant and antibacterial properties from P. crocatum, signifies the recognition of the extract as a significant reservoir of bioactive substances.

Ethnobotany, total phenolic and flavonoid content in the species Corymbia citriodora (Hook.) K.D. Hill & L.A.S. Johnson

After the emergence of COVID-19, respiratory sequelae persisted, and one alternative worth reconsideration is Corymbia citriodora (Hook.) K.D. Hill & L.A.S. Johnson. The objective is to determine the ethnobotany and the total phenolic and flavonoid content of C. citriodora. The ethnobotanical information was gathered using a semistructured questionnaire from herbalists in markets in the city of Trujillo, La Libertad, Peru. Corymbia citriodora was collected in Agallpampa, Otuzco, and identified in the Herbarium Truxillense. Three extracts were prepared and evaluated (infuse, decoct, and hydroethanolic). The total phenols were quantified using the Folin Ciocalteau method, and total flavonoids were quantified using the aluminum chloride method. Corymbia citriodora reported a higher medicinal use in bronchitis (39.1%), in the preparation of a frequent infusion, made from leaves, often in combination with Piper aduncum (35.7%). Total phenols and flavonoids were reported as 118.65 mg GAE and 33.95 mg QCE per gram of dried drug, with the hydroethanolic extract standing out. The high presence of phenolic and flavonoid bioactives is directly related to anti-inflammatory activity, suggesting that C. citriodora remains a viable option and demonstrates efficacy in a traditional context for improving respiratory conditions.

FLAVONOIDS IN ACACIA CATECHU BARK EXTRACTS AND BIBLIOGRAPHIC ANALYSIS IN GUNA, MADHYA PRADESH, INDIA

Objective: The objective of this work is to examine the overall flavonoid content in bark samples of Acacia catechu obtained from the Guna region of Madhya Pradesh, India, using several extraction methods. Furthermore, this study has incorporated current literature, conducted bibliographic analysis, examined co-authorship studies, and gathered other pertinent material to augment comprehension and underscore the importance of the research.Methods: For the purpose of this experiment, a total of thirty samples of test plants were gathered from the research region throughout various seasons. Each individual sample was prepared with six different solvents. Following this, a standardized laboratory technique was used to undertake quantitative testing. The bibliographic analysis was conducted using Dimension AI and VOS viewer software in the timeframe of 1975-2024 (till March 8, 2024).Results: Findings indicated that the polar organic solvents methanol, ethanol, acetone, and the aqueous extracts were shown to display a substantial quantity of flavonoids, chloroform extracts exhibited negligible and benzene extracts were found to be entirely devoid of the same. The comparative bibliographic investigations have confirmed the need to study the possible medicinal properties of test plants in the Guna region.Conclusion: This preliminary study has the potential to identify new, economically viable, and readily available sources of flavonoids, which are natural antioxidants present in the indigenous flora of the Guna region.

The arbuscular mycorrhizal fungi inoculation affects plant growth and flavonoid content in tomato plant (Lycopersicum esculentum Mill.)

Tomato (Lycopersicon esculentum) is one of the most important vegetables to supply nutritional needs that can be cultivated worldwide. This study was conducted to increase tomato plant growth and flavonoid content by applying arbuscular mycorrhizal fungi (AMF). The experiment used a randomized block design with six treatments of AMF concentrations, 0, 2, 4, 6, 8, and 10 g/kg of planting media. Tomato seeds were soaked in water for 1 h and then sown for 4 weeks. The seedlings with four leaves were transferred into a sterilized media previously inoculated with AMF and then were grown in a greenhouse for 10 weeks to observe growth parameters, AMF-infected root, and the total content of flavonoids. Analysis data used analysis of variance followed by the Duncan multiple range test. The result showed that AMF application into the media significantly increased the tomato plant growth parameters, AMF- infected root percentage, and flavonoid content. A 10-g AMF treatment has the best effect in this study, resulting in a plant height of 78.93 cm, stem diameter of 1.35 cm, number of leaves of 14.50, leaf area of 3861.02 cm2, leaf color scale of 3.7, plant dry weight of 1.24 g, root length of 22.42 cm, AMF-infected root of 55%, and flavonoid content of 0.053 mg/g.

Secondary metabolites from the leaves of Terminalia myriocarpa and their ?-glucosidase inhibitory potential

The objective of this study is to investigate the chemical composition of the leaves of Terminalia myriocarpa and to evaluate their ?-glucosidase inhibitory activity aiming to be used as a safe antidiabetic. Consequently, 10 compounds were isolated using column and preparative thin-layer chromatographic techniques and identified as alphitolic acid, isovitexin, flavogallonic acid, nigaichigoside F1, quercetin, quadranoside IV, rosamultin, 19?-hydroxyasiatic acid, asiatic acid, and arjunic acid. Structure elucidation was based on chemical (acid hydrolysis) and spectroscopic (UV, 1 H, and 13C-NMR) analyses and findings were confirmed through comparison with published data. The ?-glucosidase inhibitory activity of the methanol extract fractions and compounds isolated therefrom was evaluated in-vitro alongside that of acarbose (positive control). Results revealed that quercetin and flavogallonic acid were significantly active with half-maximal inhibitory concentrations (IC50) equal to 7.5 ± 0.09 and 21.0 ± 1.4 µM, respectively. Furthermore, the interaction of flavogallonic acid with ?-glucosidase enzyme was verified by docking experiment. Accordingly, quercetin and flavogallonic acid could be considered safer natural alternatives to the currently available ?-glucosidase inhibitors.

Biochemical Analysis and Characterization of Silver Nanoparticles Synthesized by the Endophytic Fungus Plectosphaerella oligotrophica S8A26 Associated with Anaphalis contorta

Endophytic fungi associated with medicinal plants produce compounds that have vast biological activities. Anaphalis contorta is a highly therapeutic plant that has been used in traditional medicine by various communities. In the present study, the endophytic fungus Plectosphaerella oligotrophica S8A26 was isolated from the stem part of A. contorta and assessed for its plant growth promotion abilities, and showed positive results for phosphate solubilization and ammonia production. The secondary metabolites were analyzed for the presence of seven biochemicals, which indicate the presence of flavonoids, phenols, saponins, and steroids. The total flavonoid content and total phenolic content were recorded to be 25.67 µg of QE/mg of EE and 5.06 µg of GAE/mg of EE, respectively. The isolate was allowed to synthesize silver nanoparticles and characterized using X-ray diffraction, Energy Dispersive X-ray, Particle size distribution, and Zeta potential. XRD analysis has shown the AgNPs are crystalline with an average size of 24.81 nm. EDX study reveals silver as the main component with 48.63%. Particle size distribution indicates that the majority of the particles occur in the size range of 65–70 nm. Zeta potential value of ?31.8 mV shows that the AgNPs are stable. Studies like the biological activities of the endophyte as well as AgNPs, secondary metabolite profiling, and the isolation of bioactive compounds would constitute important future research.

Protective effect of sterubin against neurochemical and behavioral impairments in rotenone-induced Parkinson's disease

This study was conducted to evaluate how sterubin affects rotenone-induced Parkinson's disease (PD) in rats. A total of 24 rats were distributed into 4 equal groups: normal saline control and rotenone control were administered saline or rotenone (ROT), respectively, orally; sterubin 10 received ROT + sterubin 10 mg/kg po; and sterubin alone was administered to the test group (10 mg/kg). Rats of the normal saline and sterubin alone groups received sunflower oil injection (sc) daily, 1 h after receiving the treatments cited above, while rats of the other groups received rotenone injection (0.5 mg/kg, sc). The treatment was continued over the course of 28 days daily. On the 29th day, catalepsy and akinesia were assessed. The rats were then euthanized, and the brain was extracted for estimation of endogenous antioxidants (MDA: malondialdehyde, GSH: reduced glutathione, CAT: catalase, SOD: superoxide dismutase), nitrative (nitrite) stress markers, neuroinflammatory cytokines, and neurotransmitter levels and their metabolites (3,4-dihydroxyphenylacetic acid (DOPAC), dopamine (DA), norepinephrine (NE), serotonin (5-HT), 5-hydroxyindoleacetic acid (5-HIAA), and homovanillic acid (HVA)). Akinesia and catatonia caused by ROT reduced the levels of endogenous antioxidants (GSH, CAT, and SOD), elevated the MDA level, and altered the levels of nitrites, neurotransmitters, and their metabolites. Sterubin restored the neurobehavioral deficits, oxidative stress, and metabolites of altered neurotransmitters caused by ROT. Results demonstrated the anti-Parkinson's activities of sterubin in ROT-treated rats.

Neurocognitive effects of proanthocyanidin in Alzheimer's disease: a systematic review of preclinical evidence

Cognitive disorders and dementia largely influence individual independence and orientation. Based on the Alzheimer's Disease International (ADI) estimation, approximately 75% of individuals with dementia are undiagnosed. In fact, in some low- and middle-income countries, the percentage is as high as 90%. In this systematic review, which is based on PRISMA guidelines, we aim to identify the mechanism of action of proanthocyanidin. Finding a natural product alternative as a potential nootropic can help increase the number of armamentariums against dementia and other cognitive impairments. In this preclinical research, we determined the effect of proanthocyanidins on Alzheimer's disease (AD) by searching electronic bibliographic databases like Scopus, Proquest, ScienceDirect, PubMed, and Google. There was no imposed time limit. However, the search was limited to only English articles. The review protocol is registered on PROSPERO as CRD42022356301. A population, intervention, control, and outcomes (PICO) technique was utilized for report inclusion, and all reports were assessed for risk of bias by using the SYRCLE's RoB tool. The article's bibliographic information, induction model, type of proanthocyanidins, animal strain/weight/age, and outcome measurements were acquired from ten papers and are reported here. Further analysis was validated and determined for the review. The included studies met the review's inclusion criteria and suggested that proanthocyanidins have a neurocognitive effect against AD. Additionally, the effectiveness of proanthocyanidins in reducing oxidative stress, acetylcholinesterase activity, amyloid beta, its efficacy in alleviating superoxide dismutase, cognitive properties, and in facilitating cholinergic transmission in various models of AD has been collectively observed in ten studies.

Inhibitory effect and mechanism of total flavonoids from Melicope pteleifolia on transplanted tumor of colorectal cancer in nude mice / 中国药房

OBJECTIVE To study the inhibitory effect and mechanism of total flavonoids from Melicope pteleifolia （TF-MPL） on transplanted tumor of colorectal cancer in nude mice. METHODS The transplanted tumor model of colorectal cancer was induced by injecting 0.2 mL colorectal cancer cell LoVo subcutaneously via the right armpit of nude mice. After successful modeling， nude mice were randomly divided into model group， 5-fluorouracil group （positive control， 10 mg/kg）， TF-MPL high- dose and low-dose groups （25， 12.5 mg/kg）； a normal group （normal saline containing 0.3% carboxymethyl cellulose sodium） without modeling was additionally set up， with 6 mice in each group. Each group was intraperitoneally injected with the corresponding drug solution/solvent for 21 consecutive days. The inhibitory rate of the transplanted tumor， liver and spleen index， and the levels of tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6） in serum were detected after the last medication； the morphological changes of tumor tissue were observed； immunohistochemical staining was used to detect protein expressions of Toll- like receptor 4 （TLR4） and nuclear factor-κB subunit p65 （NF-κB p65） in tumor tissue of nude mice. Western blot assay was used to detect protein expressions of TLR4， myeloid differentiation factor 88 （MyD88）， TNF receptor-associated factor 6 （TRAF6）， interleukin-1 receptor-associated kinase 1 （IRAK-1）， NF-κB p65 and caspase-3 in tumor tissue of nude mice. RESULTS Compared with the model group， TF-MPL high-dose group showed a significant decrease in tumor weight （inhibitory rate of 36.91%）， liver and spleen index， serum levels of TNF-α and IL-6 and protein expressions of TLR4， MyD88， TRAF6，IRAK-1 and NF- κB p65 （P＜0.05 or P＜0.01）； the expression of caspase-3 protein was increased significantly （P＜0.05）， and more tumor cell shrinkage and deformation， nuclear pyknosis and fragmentation were observed. CONCLUSIONS TF-MPL can significantly inhibit the growth of transplanted tumor of colorectal cancer in nude mice， the mechanism of which may be associated with reducing inflammatory response， inhibiting TLR4/MyD88/NF-κB signaling pathway， and promoting apoptosis in colorectal cancer cells.

Quantitative Analysis of Multi-components in Gnaphalium affine Based on UPLC-Q-Exactive Orbitrap MS / 中国实验方剂学杂志

ObjectiveTo establish a rapid and stable liquid chromatography-mass spectrometry（LC-MS） for simultaneous analysis of 17 chemical components in Gnaphalium affine aboveground parts with flowers， so as to provide experimental basis for improving the quality standard of this herb. MethodUltra performance liquid chromatography-quadrupole/electrostatic field orbitrap mass spectrometry（UPLC-Q-Exactive Orbitrap MS） was used for the quantitative analysis of 17 constituents in 15 batches of G. affine from different origins， the separation was performed on an ACQUITY UPLC® BEH C18 column（2.1 mm×100 mm， 1.7 μm） with the mobile phase of methanol（A）-0.1% formic acid aqueous solution（B） for gradient elution（0-1.0 min， 8%A； 1.0-4.0 min， 8%-26%A； 4.0-9.0 min， 26%A； 9.0-14.0 min， 26%-34%A； 14.0-14.5 min， 34%-45%A； 14.5-15.0 min， 45%-60%A； 15.0-18.0 min， 60%-90%A； 18.0-19.0 min， 90%A； 19.0-19.01 min， 90%-8%A； 19.01-20.0 min， 8%A）， the flow rate was 0.3 mL·min-1， the column temperature was 40 ℃ and the injection volume was 2 μL. And the electrospray ionization was used with full scanning in both positive and negative ion modes， and the scanning range was m/z 100-1 000. ResultThe established method has been verified by the methodology and could be used for the simultaneous quantification of 17 components in G. affine. The content ranges of the 17 components（quinic acid， gallic acid， protocatechuic acid， neochlorogenic acid， chlorogenic acid， cryptochlorogenic acid， caffeic acid， 1，3-O-dicaffeoylquinic acid， isochlorogenic acid A， isoquercitrin， 1，5-O-dicaffeoylquinic acid， apigenin-7-O-glucoside， astragalin， isochlorogenic acid C， luteolin， apigenin and hispidulin） in 15 batches of G. affine samples was 39.60-179.12， 0.17-0.84， 2.41-8.38， 4.33-31.50， 13.63-180.38， 2.43-14.75， 1.16-19.68， 0.49-5.63， 55.77-445.16， 0.23-10.26， 62.04-530.10， 1.11-18.01， 11.36-90.61， 12.22-65.98， 7.22-69.84， 3.37-45.65， 0.30-2.59 μg·g-1， respectively. The content of organic acids was higher than that of flavonoids in G. affine， and the contents of 1，5-O-dicaffeoylquinic acid， isochlorogenic acid A， quinic acid and chlorogenic acid were higher. Meanwhile， the content of flavonoids in the samples from Guizhou was higher than that from Jiangsu， while the content of organic acids in the samples from Jiangsu was higher than that from Guizhou. ConclusionThe established method can be used for the rapid and accurate determination of 17 components in G. affine， which clarifies the content range of the main components in this herb， and can provide a reference for the selection of quality control markers of G. affine.

Optimization of extraction process of flavonoids from Broussonetia papyrifera leaves and its antioxidant effect on mouse epidermal stem cells / 中国生物制品学杂志

@#Objective To optimize the extraction process of flavonoids from Broussonetia papyrifera leaves and explore the antioxidant effect of flavonoids on mouse epidermal stem cells.Methods The extraction process of flavonoids from Broussonetia papyrifera leaves was optimized by single factor experiment,including the liquid-solid ratio(15:1,20:1,25:1,30:1and 35:1),sodium hydroxide(NaOH) concentration(0.2%,0.4%,0.6%,0.8% and 1.0%),pH value(2.5,3.0,3.5,4.0and 4.5) and extraction temperature(60,65,70,75 and 80℃).Based on the results of single factor experiment,the optimal extraction process was determined by orthogonal test with the mass fraction of flavonoids as the evaluation index.CD49f~+/CD71~-mouse epidermal stem cells were isolated and cultivated by immunomagnetic bead method,and the effects of flavonoids on the cell relative viability and the contents of reduced glutathione(GSH) and malondialdehyde(MDA) were detected.Results The optimal extraction conditions of flavonoids were liquid-solid ratio of 30:1,0.6% NaOH,pH 4.5and extraction temperature of 75 ℃.Under these conditions,the average mass fraction of flavonoids extracted was 1.47%.Compared with the negative control group,when the flavonoids final concentration was 25 and 50 μg/mL,the cell relative viability increased significantly(F=1.427 and 13.747 respectively,each P <0.01);when the final concentration of flavonoids was 12.5,25 and 50 μg/mL,the content of GSH increased significantly(F=0.044,0.291 and 2.577 respectively,each P <0.05) and the content of MDA decreased significantly(F=3.568,4.909 and 1.400 respectively,each P <0.05).Conclusion The optimized extraction process of flavonoids from B.papyrifera leaves was stable and reliable,which is beneficial to the reuse of remaining stock solution after processing,and the extracted flavonoids can promote the proliferation of mouse epidermal stem cells and perform antioxidant activity.

Evaluation of Relationship of Pueraria lobata Quality from Different Producing Areas Based on "Climate Factor-component Content-antioxidant Capacity" / 中国实验方剂学杂志

ObjectiveThe correlation of Pueraria lobata producing areas， climate factors， total flavonoids of P. lobata， polysaccharide content of P. lobata， and antioxidant activity of P.lobata for medicinal application was analyzed， and the relationship between climate factors and the formation of P. lobata quality was evaluated. MethodThe scavenging rates of 1,1-diphenyl-2-picrylhydrazyl radical 2,2-diphenyl-1-(2,4,6-trinitrophenyl)hydrazyl（DPPH） and 2, 2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)（ABTS） radicals by total flavonoids and polysaccharides of P. lobata were detected， and the correlation between the contents of each component and the information of producing areas and climate factors was analyzed. ResultThe ABTS+ scavenging rate by total flavonoids of P.lobata was negatively correlated with altitude （P<0.05） and positively correlated with annual sunshine hours （P<0.05）. The altitude was positively correlated with the total flavonoid content， while the annual sunshine hours were negatively correlated with the total flavonoid content. There was a negative correlation between total flavonoid content and ABTS+ scavenging rate by total flavonoids. In other words， lower altitude and longer annual sunshine hours indicated lower total flavonoid content and higher ABTS+ scavenging rate by total flavonoids. The ABTS+ scavenging rate by polysaccharides of P. lobata was negatively correlated with the frost-free period （P<0.05） and the mean temperature in July （P<0.01）. There was a positive correlation between the polysaccharide content of P. lobata and the frost-free period. The mean temperature in July was positively correlated with the polysaccharide content of P. lobata （P<0.05）. The polysaccharide content of P. lobata was negatively correlated with the ABTS+ scavenging rate by polysaccharides of P. lobata. In other words， a shorter frost-free period in the producing area and lower mean temperature in July indicated lower polysaccharide content of P. lobata and higher ABTS+ scavenging rate by polysaccharides of P. lobata. The mean temperature in July was significantly correlated with the contents of total flavonoids and polysaccharides in P. lobata samples （P<0.05）. The lower mean temperature in July was often accompanied by lower total flavonoid content of P. lobata， lower polysaccharide content of Pueraria lobata， and stronger antioxidant activity of P. lobata samples. ConclusionThe ability of P. lobata to remove ABTS+ is stronger than that of DPPH+. There is a significant correlation between climate factors， content， and antioxidant capacity in each producing area. Further research on the internal law of the formation of medicinal active components of P. lobata induced by core climate factors will provide a scientific basis for revealing the formation mechanism of genuine P. lobata and the subsequent control of P. lobata quality according to the environment of producing areas.

Analysis of Differences in Secondary Metabolites from Dendrobium nobile Stems Cultivated on Epiphytic Rocks in Different Growth Years Based on Metabolomics / 中国实验方剂学杂志

ObjectiveTo study the changing characteristics of secondary metabolic compounds accumulated in Dendrobium nobile stems at different growth years， a simulated wild stone plant， in order to provide a theoretical basis for rational planning of the harvesting period of D. nobile. MethodUltra-high performance liquid chromatography-mass spectrometry（UPLC-MS/MS） was used to detect and analyze the secondary metabolites in the stems of 1-year-old， 2-year-old， and 3-year-old D. nobile. The mass spectrometry data were processed using Analyst 1.6.3 software， and all samples were subjected to principal component analysis（PCA）， cluster heat map analysis， partial least squares-discriminant analysis（PLS-DA）， and differential secondary metabolites were screened based on variable importance in projection（VIP） values>1， fold change（FC）≥2 and FC≤0.5. Then differential secondary metabolites were identified based on relative molecular weight， fragmentation ions and mass spectrometry database， and enriched pathways were identified based on the Kyoto Encyclopedia of Genes and Genomes（KEGG） database. ResultA total of 1 317 secondary metabolites were identified in the stems of D. nobile at three growth stages， with flavonoids， phenolic acids， alkaloids and terpenoids accounting for 76.55% of the total. Compared with the 1-year-old stems of D. nobile， 289 differential secondary metabolites were identified in the 2-year-old stems， of which 255 were up-regulated and 34 were down-regulated， 682 differential secondary metabolites were identified in the 3-year-old stems， of which 502 were up-regulated and 180 were down-regulated. Compared to the 2-year-old stems， the 3-year-old stems had 602 differential secondary metabolites， with 405 up-regulated and 197 down-regulated. As the growth stage of D. nobile increased， the top 10 up-regulated differential metabolites mainly included flavonoids， phenolic acids， phenylpropanoids and terpenoids， such as kaempferol derivatives， asperulosidic acid， apigenin derivatives， chrysoeriol derivatives， isorhamnetin derivatives， taxifolin derivatives， quercetin derivatives. KEGG enrichment analysis showed significant enrichment of secondary metabolites in the flavonoid biosynthesis， flavone， and flavonol biosynthesis， secondary metabolite biosynthesis， and phenylpropanoid biosynthesis pathways with the increase of growth years. ConclusionWith the increase of the growth years， the levels of secondary metabolites such as flavonoids， phenolic acids， phenylpropanoids and terpenoids in the wild-grown D. nobile have been significantly enhanced. In practical production， grading based on different growth years can be carried out to improve the medicinal and economic values of D. nobile.

Role of Flavonoids in Traditional Chinese Medicine to Improve Cerebral Ischaemia-reperfusion Injury and Its Mechanism： A Review / 中国实验方剂学杂志

Cerebral ischemia-reperfusion injury （CIRI） has a very high incidence， disability， and mortality rates， which seriously affects human life and health. In recent years， modern medicine has made some progress in the diagnosis and treatment of CIRI， but there are still problems such as difficulties in postoperative rehabilitation and adverse drug reactions， and new therapeutic drugs for CIRI are urgently needed. As an important class of active ingredients in traditional Chinese medicine， flavonoids can play antioxidant， apoptosis inhibition， anti-inflammatory， and other pharmacological effects to improve brain tissue damage， which is important for improving the quality of life of CIRI patients and slowing down the aging of the social population. Numerous studies have found that flavonoids in traditional Chinese medicine can regulate cell surface receptors Toll-like receptor 4/nuclear factor-kappaB （TLR4/NF-κB）， phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt）， adenylate-activated protein kinase/mammalian target of rapamycin protein （AMPK/mTOR）， Ras homologous gene family member A/Rho-associated coiled-coil protein kinase （RhoA/ROCK）， nuclear factor E2-associated factor 2/Kelch-like epoxychloropropane-associated protein-1/haemoglobin oxygenase 1 （Nrf2/Keap1/ HO-1）， Notch， and other signaling pathways， so as to regulate the transcription and expression of related proteins after CIRI， alleviate brain tissue injury， and improve CIRI. This paper analyzed the relevant literature in China and abroad in recent years， reviewed the mechanism of action and related pathways of flavonoids in traditional Chinese medicine to improve CIRI， and explored the new therapeutic direction of CIRI at the metabolic level， with a view to providing a basis for the further development and application of flavonoids in traditional Chinese medicine.