D101 Macroporous Resin Research Articles

Extraction and Purification of Flavonoids and Antiviral and Antioxidant Activities of Polygonum perfoliatum L.

The aim of the present study was to optimize the process parameters for the extraction and purification of total flavonoids from Polygonum perfoliatum L., in addition to analyzing their chemical composition and evaluating their activity against varicella-zoster virus (VZV) and antioxidant activity. The optimum extraction process was determined using one-way and response surface methods with the following conditions: ethanol concentration of 82.00%, temperature of 90.29 °C, solid-to-liquid ratio of 1:32.78 g/mL, extraction time of 1.5 h, and two extractions with a yield of 14.98 ± 0.11 mg/g. Purification was then carried out using D101 macroporous resin to obtain a flavonoid purity of 43.00 ± 2.55%, which was 3.38 times higher than that of the crude extract (12.74 ± 1.04%). Further purification was carried out using a 1:9 hybrid column of macroporous resin and polyamide, and the purity of flavonoids was enhanced to 59.02 ± 2.23%, which is 1.37 times higher than that of the macroporous resin purifier (43.00 ± 2.55%) and 4.63 times higher than that of the crude extract (12.74 ± 1.04%). Seventy-nine flavonoids were identified using ultra-performance liquid chromatography-tandem high-resolution mass spectrometry (UPLC-HRMS). In addition, the purified flavonoids showed good anti-VZV and antioxidant activities. Therefore, this study can provide technical support and theoretical basis for the further development and utilization of Polygonum perfoliatum L. resources.

Isolation and purification of polyphenols, hypoglycemic and hypolipidemic and active constituent analysis of walnut septum polyphenols

Walnut septum is rich in polyphenolic substances, but the hypoglycaemic and hypolipidemic effects of walnut septum polyphenols (WSP) and the main active compounds responsible for these effects are still unknown. In the present study, WSP was firstly extracted, and then purified using D101 macroporous resin, and the purity of WSP obtained was 87.36 %. WSP inhibited the activity of α-glucosidase and pancreatic lipase with IC50 values of 36.2 μg/mL and 155.9 μg/mL, respectively. Furthermore, WSP increased glucose consumption and decreased oxidative stress in insulin-resistant C2C12 cells, as well as inhibited lipid accumulation in 3 T3-L1 adipocytes. Eight naturally active ligands for α-glucosidase and pancreatic lipase were identified from WSP by affinity ultrafiltration combined with liquid chromatography-mass spectrometry analytical methods. Further molecular docking showed strong binding between these eight ligands and the enzyme. The results of the study lay a scientific foundation for the development of walnut septum as a functional food for hypoglycemic and hypolipidemic.

Preparation of polyacylated anthocyanins of red radish and enhancing its stability and bioaccessibility through encapsulating within double-coated yeast glucan particles

Abstract Although the anthocyanins of red radishes (ARR) rich in polyacylated pelargonidin glucosides are used as commercial food pigment, they are unstable. We found that the anthocyanin purity of the crude ARR extract can be rapidly increased by 5 times using D101 macroporous resin (mass ratio of extract:resin = 1:2.2). Capsule constructed by chitosan (CTS), pectin (PT) and yeast glucan particles (YGP) presented a high efficiency to encapsulate ARR. Liquid chromatography-mass spectrometry analysis verified that polyacylated pelargonidin glucosides in ARR were packed into the capsule. Microstructure observation and Fourier transforms infrared spectroscopy further confirmed the encapsulated structure. Co-encapsulation of CTS, PT and YGP showed effective protection for ARR against heat, oxygen, ascorbic acid, and physiological pH. This encapsulation also significantly improved the gastric and intestinal bioaccessibilities of ARR. These results suggested that the triplex-coated YGPs might be a promising strategy to protect and deliver polyacylated anthocyanin.

Extraction, in vitro hypoglycaemic activity and active ingredient analysis of polyphenols from walnut green husk

A variety of polyphenols in walnut green husk have shown physiological benefits, but their primary hypoglycemic active ingredients remain unclear. We report the extraction of walnut green husk polyphenols (WGHP) and their hypoglycemic effects in vitro. WGHP was extracted using ultrasound-assisted ethanol extraction, with optimization via response surface methodology resulting in 88.0 ± 0.1 % purity after D101 macroporous resin purification. WGHP effectively inhibited α-glucosidase and α-amylase, with IC50 values of 3.42 μg/mL and 2.56 mg/mL, respectively. It also increased glycogen synthesis in insulin-resistant HepG2 cells and enhanced glucose consumption in 3T3-L1 and insulin-resistant HepG2 cells. Screening six α-glucosidase ligands from WGHP using affinity ultrafiltration and UPLC-MS/MS, combined with molecular docking, identified hydrogen bonds and binding energies between the ligands and the enzyme. These results highlight the hypoglycemic potential of walnut green husk polyphenols and provide a basis for future therapeutic research.

Elucidating the eco-friendly herbicidal potential of microbial metabolites from Bacillus altitudinis.

Microbial herbicides play a vital role in agricultural preservation, amid growing concerns over the ecological impact from extensive development and use of chemical herbicides. Utilizing beneficial microbial metabolites to combat weeds has become a significant focus of research. This study focused on isolating herbicidal active compounds from Bacillus altitudinis D30202 through activity-guided methods. First, the n-butanol extract (n-BE) of B. altitudinis D30202 underwent fractionation using macroporous adsorption resin D101 and Sephadex LH-20, identifying Fr. F as the most potent segment against wild oats (Avena fatua L.). Ultra-performance liquid chromatography - quadrupole time-of-flight mass spectrometry (UPLC - QTOF-MS) identified nine compounds in the active fraction Fr. F. Subsequently, three subfractions (Fr.F-1 to Fr.F-3) were derived from Fr.F via semi-preparative liquid chromatography, resulting in methyl indole-3-acetate (MeIAA) purification. MeIAA, functioning as an auxin analog, exhibited effects of indole-3-acetic acid (IAA) on wild oats' growth, with a root length median inhibitory concentration of 81.06µg/ml. Furthermore, we assessed MeIAA's herbicidal impact on five weed species across diverse families and genera, providing a first-time analysis of MeIAA's mechanism on wild oats. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) revealed structural damage to leaves and roots post-MeIAA treatment. MeIAA treatment increased superoxide anion and hydrogen peroxide levels in wild oat roots, alongside with elevated peroxidase (POD) and superoxide dismutase (SOD) activity, chlorophyll-degrading enzymes (Chlase, MDACase), malondialdehyde (MDA) content, and relative conductivity in leaves. Conversely, it decreased catalase (CAT) activity and chlorophyll content. Therefore, this study provides a new material source and theoretical foundation for ecologically sustainable agricultural weed control.

Isolation and purification of flavonoids from quinoa whole grain and its inhibitory effect on lipid accumulation in nonalcoholic fatty liver disease by inhibiting the expression of CD36 and FASN.

Nonalcoholic fatty liver disease (NAFLD), a chronic metabolic disorder marked by excessive lipid deposition, represents a considerable health burden with no established efficacious treatment strategy. Quinoa (Chenopodium quinoa Willd.), valued for its health benefits, is replete with flavonoid bioactives. The aims of this work are to isolate and purify flavonoids from quinoa whole grain that can intervene in NAFLD and to elucidate some of the underlying mechanisms. Chenopodium quinoa Willd. flavonoids (CQWF) were obtained successfully through an optimized ultrasonic extraction methodology, followed by isolation and purification utilizing macroporous resin D101. The study then explored the therapeutic potential of CQWF and its eluted fractions in models emulating NAFLD conditions: an in vitro fatty liver cell model induced by oleic acid (OA) and palmitic acid (PA) in the HepG2 and BEL-7402 cell lines, and an in vivo high-fat diet (HFD)-induced NAFLD model in C57BL/6N mice. The findings revealed a comprehensive mitigating effect of CQWF30 on NAFLD, manifesting in reduced intracellular lipid accumulation in steatotic hepatocytes and a concerted downregulation of key lipid metabolism genes, CD36 and FASN. Administration of CQWF30 reduced triglyceride (TG) levels in both the cellular model and the livers of HFD-fed mice. It also reduced serum concentrations of TG, total cholesterol (T-CHO), low-density lipoprotein cholesterol (LDL-C), aspartate aminotransferase (AST), and alanine aminotransferase (ALT), while increasing high-density lipoprotein cholesterol (HDL-C) in the mice. These results highlighted the promising therapeutic capacity of CQWF, particularly CQWF30. This research advances the exploration and utilization of flavonoids derived from quinoa whole grain, providing innovative dietary intervention strategies for NAFLD. © 2024 Society of Chemical Industry.

Optimization and evaluation of biologically active compound with antibacterial and anti-inflammatory properties from peony seed meal

Peony seed meal is produced annually on a large scale in China, resulting in considerable solid residue. Unfortunately, most of this residue is discarded, thereby creating environmental pressure. However, the untapped medicinal value of peony seed meal prompted us to establish an efficient methodology for optimizing the extraction and purification of total alkaloid, followed by a detailed analysis of its biological activities. A single-factor experiment and an orthogonal test were used to identify the optimal extraction parameters. Then, further optimization was performed using the response surface methodology. The extract was enriched using D-101 macroporous adsorption resin. Subsequently, silica gel column chromatography was used for separation, and high-performance liquid chromatography was used for further separation and quantification. Meanwhile, gas chromatography–mass spectrometry (GC–MS) was used to identify alkaloid components. Finally, the antibacterial and anti-inflammatory were evaluated. The optimal extraction conditions substantially improved the yield of alkaloids. A total of 15 alkaloids were separated. GC–MS analysis revealed the presence of Sophoridine, which accounted for 91.88 % of the total alkaloid content. The antibacterial activity of the total alkaloid was tested, and the lowest inhibitory concentrations of E. coli ATCC 25922 and S. aureus ATCC 29213 were 0.04516 and 0.05645 mg/mL, respectively. Additionally, total alkaloid had an inhibitory effect on ear swelling induced by xylene and toe swelling induced by egg white, reducing by 80 % and 24 %, respectively. The current research discovered the antibacterial activity of PSM alkaloid, which has important practical significance for the research and development of new plant-derived antibacterial agents.

Potential active fractions and constituents in the flowers of Trollius chinensis Bunge responsible for treating acute pharyngitis

Ethnopharmacological relevanceTrollius chinensis Bunge has a long history of use in China as traditional Chinese medicine and functional tea for the treatment of respiratory infections, such as pharyngitis, tonsillitis and bronchitis. Pharyngitis can impact the entire throat and adjacent lymphoid tissues, and may lead to significant systemic complications. However, the active components and mechanism of Trollius chinensis Bunge for treating acute pharyngitis remains unclear. Aim of the studyTrollius chinensis Bunge is recognized in China both as a medicinal herb and a functional tea. Research into its properties aimed to establish its effectiveness against pharyngitis and to pinpoint the active components and mechanism. Materials and methodsA 70% ethanol extract from the herb was prepared, which was refined using chromatography through a column containing D101 macroporous resin and varying ethanol solutions. The efficacy of the initial and refined extracts was tested using a rat model of ammonia-induced acute pharyngitis. Pathological examination, HE staining and ELISA were applied to screen activity fraction. The compounds were isolated by silica gel, sephadex LH-20 column chromatography and semi-preparative HPLC chromatography from active fraction. All of the isolated compounds were assessed for anti-inflammatory activity by acting on LPS-induced RAW 264.7 macrophage cells in vitro. Cytotoxicity of compounds was detected by CCK-8 assay. The Griess reaction was applied to evaluate the inhibitory effects of isolated compounds on NO production in RAW 264.7 cells induced by LPS. TNF-α, IL-6, IL-1β and PGE2 levels in macrophage supernatant were detected by ELISA. Molecular docking and western blot analysis were applied to study the anti-inflammatory mechanism of active compound. ResultsThe fraction extracted with 30% ethanol proved particularly effective, significantly reducing pharyngitis symptoms. This was evidenced by decreased levels of cytokines (TNF-α, IL-6, IL-1β and PGE2) and visible improvements in the pharyngeal tissue histology. In pursuit of pharyngitis treatments, 23 phenolic acids and 13 flavonoids were isolated from the 30% ethanol fraction and identified using spectral analysis. Of these, three were newly discovered compounds and eight were first-time isolates from the Trollius genus. These compounds were further investigated for their ability to suppress nitric oxide production in RAW 264.7 cells triggered by lipopolysaccharide. Compounds 3, 19, and 26 exhibited strong anti-inflammatory properties. HPLC analysis of the 30% ethanol fraction revealed that orientin was the predominant component, accounting for 44.4% of this fraction. Western blot analysis demonstrated that orientin reduced the expression levels of the protein p-p65 relative to p65, p-IκBα relative to IκBα and iNOS, indicating an anti-inflammatory effect potentially through the modulation of the NF-κB signaling pathway. ConclusionThe finding of this study provided strong support for the use of T. chinensis as a potential functional food for treating pharyngitis.

Extraction effects of eight deep eutectic solvents on dianhong black tea: From chemical composition analysis to antioxidant and α-glucosidase inhibitory assessments

Deep eutectic solvent (DES) has been widely developed and applied in extracting the functional components of teas, however, few studies devoted to revealing the composition differences of tea extracts obtained with DES. The present study aims to study the chemical composition and bioactive differences of Dianhong congou black tea extracts prepared by eight choline chloride (ChCl)-based DES. In addition, D-101 macroporous resin was used to remove DES for bioactivity analysis. High performance liquid chromatography (HPLC) and LC-mass spectrometry (LC-MS) were applied to quantitatively and qualitatively analyze chemical composition differences of DES extracts before and after DES removal. Quantitative results showed that DES4 (composed of ChCl and ethylene glycol) was suitable for extracting catechins (e.g., the extracted level of epicatechin gallate was 8.75 mg/g), theaflavins (theaflavin-3,3′-digallate, 1.59 mg/g), and monosaccharides (glucose, 4.97 mg/g), meanwhile DES5 (composed of ChCl and glycerol) was proved to be effective for extracting free amino acids (with highest level of theanine, 19.72 mg/g). LC-MS based metabolomics results showed that DES2 (ChCl-citric acid), DES3 (ChCl-proline), and DES4 had considerable remaining amounts of flavonoid glycosides, theasinensins, and hydrolysable tannins after DES removal. Although purified DES5 extract contained a comparable amount of these marker compounds to other purified DES extracts, its antioxidant and α-glucosidase inhibitory activities were the lowest among others. Combining results from above, DES4 was proved to be the optimum solvent for extracting functional components of Dianhong congou black tea. The finding provided basis for the application of DES extraction method in tea science.

Ultrasound-assisted deep eutectic solvents extraction of podophyllotoxins from Juniperus sabina L: Optimization, enrichment, and anti-drug resistant bacteria activity

Podophyllotoxin, along with its congeners and derivatives exhibit strong biological activity mainly as antitumor drugs and antiviral agents. Here, the ultrasound-assisted extraction of podophyllotoxins including α-peltatin, podophyllotoxin, podophyllotoxinone and deoxypodophyllotoxin from Juniperus sabina L. leaves using deep eutectic solvents (DESs) were developed first time. To simultaneously and effectively extract four target podophyllotoxins, 19 DESs were screened to determine the best solvent to extract podophyllotoxins. Choline chloride-lactic acid (1:1) with 30 % water content was proved to have the best extraction effect. The optimal extraction process with DES were DES/solid ratio 30 mL/g, reaction temperature 44℃, time 53 min and ultrasound power 495 W. Under optimized extraction conditions, the total podophyllotoxins extraction yield was 27.520 mg/g, 196.84 % higher than that using traditional methanol solvent. The yield of α-peltatin, podophyllotoxin, deoxypodophyllotoxin and podophyllotoxinone were 13.567, 9.548, 2.021, 2.204 mg/g, respectively. Subsequently, the podophyllotoxins were effectively isolated from the deep eutectic solvent using D101 macroporous resin. The optimal process for enriching target podophyllotoxins by D101 macroporous resin were as follows: 30 mg/mL of crude extract solution, five bed volumes feed volume for adsorption; six bed volumes of 80 % ethanol for desorption. Moreover, a scale-up enrichment experiment of podophyllotoxins was performed under optimum procedure, and the results showed that the target podophyllotoxins yield was increased by 8.92 times, with recoveries in the range of 75.86–85.69 %. Additionally, the enriched podophyllotoxins effectively restored the antibacterial efficacy of cefazolin against methicillin-resistant Staphylococcus aureus and colistin-resistant Escherichia coli, resulting in 16–64-fold reduction in MIC. Therefore, the method developed in this study could be an eco-friendly, efficient and sustainable alternative for the selective extraction and enrichment of active compounds from plant material.

Comprehensive Screening and Characterization of α-glucosidase Inhibitory Components in the Edible Medicinal Plant Pholidota cantonensis Rolfe Using UPLC-Q-TOF-MS/MS Analysis and Molecular Docking.

Pholidota cantonensis Rolfe is an edible medicinal plant in the genus Pholidota of the family Orchidaceae. This plant is used to prepare medicated food in China and has been reported to possess anti-α-glucosidase activity. To date, little is known about the active substances responsible for the observed anti-α-glucosidase activity. In the present study, we aimed to screen and characterize the α-glucosidase inhibitory fraction of P. cantonensis using ultraperformance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS/MS) analysis and molecular docking. As a result, the 50% ethanol fraction obtained from D101 macroporous adsorption resin column chromatography (D50 fraction) had the highest total phenol content (353.83 ± 6.06mg GAE/g) and the most prominent α-glucosidase inhibitory activity (IC50 = 30.01 ± 7.30µg/mL). Forty-five compounds were identified from the D50 fraction by using UPLC-Q-TOF-MS/MS analysis. Molecular docking results showed that six main constituents, namely, crepidatin, 2,7-dihydroxy-4-methoxyl-9,10-dihydrophenylene, 4,4',5,6-tetrahydroxystilbene, 4,7-dihydroxy-2-methoxyl-9,10-dihydrophenylene, (-)-lariciresinol, and thunalbene, in the D50 fraction occupied the catalytic sites of α-glucosidase through strong hydrophobic interactions, hydrogen bonding, and other patterns. The binding energies were between - 29.95 and - 11.41kJ/mol, indicating good binding between the tested compounds and α-glucosidase. The active ingredients responsible for the α-glucosidase inhibitory activity may include phenanthrenes, stilbenes, dibenzyls, and lignans. The D50 fraction has potential value for developing innovative drugs for the prevention and treatment of diabetes mellitus (DM) and is worthy of in-depth research.

Screening and characterization of cosmetic efficacy components of Terminalia chebula based on biological activity-guided methodology.

Terminalia chebula exhibits a high level of antioxidant capacity and is highly valued in medicine and cosmetics. However, its main efficacy and active ingredients related to antioxidant, whitening, and anti-aging are still unclear. In this study, the active site responsible for its cosmetic efficacy was specified by the biological activity-guided method and further characterized by using ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS/MS). T.chebula was ultrasonically extracted by five solvents, and 30% ethanol extract was screened out for subsequent purification by 1,1-D-iphenyl-2-picrylhydrazyl radical (DPPH), 2,2'-Azinobis-(3-ethylbenzothiazoline-6-sulphonate) (ABTS), hydroxyl, and superoxide anion free radical scavenging assays. Five elution fractions were obtained by column chromatography on D101 macroporous adsorbent resin eluted by an increased proportion of ethanol. The 30% ethanol elution fraction was specified as the enrichment site of active ingredients showing good antioxidant capacity and potent inhibitory activity against tyrosinase and elastase. A total of 30 compounds were identified by UHPLC-QTOF-MS/MS in the 30% ethanol elution fraction, including 11 gallotannins, 14 ellagitannins, and 5 other compounds, and these compounds may be the key ingredients in cosmetics beneficial for the skin. Such a biological activity-guided method has provided a simple and rapid venue for specifying the components of medicinal herbs responsible for cosmetic efficacy.

A green and simple method for enrichment of major diterpenoids from the buds of Wikstroemia chamaedaphne with macroporous resins and their activation of latent human immunodeficiency virus activity

In this study, a green and efficient enrichment method for the four majors active diterpenoid components: pimelotide C, pimelotide A, simplexin, and 6α,7α-epoxy-5β-hydroxy-12-deoxyphorbol-13-decanoate in the buds of Wikstroemia chamaedaphne was established using macroporous resin chromatography. The adsorption and desorption rates of seven macroporous resins were compared using static tests. The D101 macroporous resin exhibited the best performance. Static and dynamic adsorption tests were performed to determine the enrichment and purification of important bioactive diterpenoids in the buds of W. chamaedaphne. Diterpenoid extracts were obtained by using D101 macroporous resin from the crude extracts of W. chamaedaphne. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis demonstrated that most of the diterpenoids were enriched in diterpenoid extracts. These results confirmed that diterpenoids in the buds of W. chamaedaphne could be enriched using macroporous resin technology, and the enriched diterpenoid extracts showed more efficient activation of the latent human immunodeficiency virus. This study provides a novel strategy for discovering efficient and low-toxicity latency-reversing agents and a potential basis for the comprehensive development and clinical application of the buds of W. chamaedaphne.

Isolation and identification of malonyl ginsenoside-Ra_1 and malonyl ginsenoside-Ra_2 from fresh roots of Panax ginseng

The aim of this paper is to study the malonyl ginsenosides in the fresh roots of Panax ginseng. D101 macroporous adsorption resin, ODS, and preparative HPLC were employed to separate the chemical components from the 70% ethanol extract of the fresh roots of P. ginseng, and the structures of the separated compounds were identified based on the data of high-resolution mass spectrometry and nuclear magnetic resonance spectroscopy. Two malonyl ginsenosides were isolated from the fresh roots of P. ginseng and identified as 3-O-\[6-O-malonyl-β-D-glucopyranosyl-(1→2)-β-D-glucopyranosyl\]-20-O-\[ β-D-xylopyranosyl-(1→4)-α-L-arabinopyranosyl-(1→6)-β-D-glucopyranosyl\]-dammar-24-ene-3β,12β,20S-triol(1) and 3-O-\[6-O-malonyl-β-D-glucopyranosyl-(1→2)-β-D-glucopyranosyl\]-20-O-\[ β-D-xylopyranosyl-(1→2)-α-L-arabinofuranosyl-(1→6)-β-D-glucopyranosyl\]-dammar-24-ene-3β,12β,20S-triol(2), respectively. Compounds 1 and 2 are new compounds isolated from fresh roots of P. ginseng for the first time and named as malonyl ginsenoside-Ra_1 and malonyl ginsenoside-Ra_2, respectively.

Silk fibroin microspheres loaded Rehmannia Liuwei extract for the protection of endothelial cells from the inhibitory effects

Liuwei Dihuang (LWDH) is a multi-component and multi-target Chinese herbal compound widely used for treating chronic conditions such as diabetes, diabetic nephropathy, hypertension, osteoporosis, and chronic kidney disease. However, traditional Chinese medicine (TCM) preparations like decoction and pill face limitations, including low active component concentration, limited bioavailability, short half-life, and the need for high dosage, which may increase the burden on liver and kidney functions and reduce clinical efficacy. In this study, LWDH was further purified using D101 macroporous adsorption resin, resulting in a soluble extract with an active component content 53.6 times higher than that of LWDH itself. The freeze-dried LWDH extract was then encapsulated within silk fibroin (SF) microspheres to significantly enhance the sustained release performance of the drug. In a human umbilical vein endothelial cell (HUVEC) model cultured under high glucose conditions, methanol vapor-treated SF/LWDH microspheres demonstrated a decrease in the 24-hour drug release rate from 61.88 % to 34.81 %, augmenting their protective effect on endothelial cells.

Impact of Liparis nervosa extract on neuroinflammation mediated by LPS-induced BV-2 microglial cells and its bioactive components analysis

To investigate the impact and potential mechanisms of extracts from different parts of Liparis nervosa on neuroinflammation by lipopolysaccharide(LPS)-induced BV-2 microglial cells. The materials of L. nervosa were subjected to crushing, ethanol extraction, and concentration to obtain an alcohol extract. Subsequently, the extract was further extracted to obtain petroleum ether extract, ethyl acetate extract, N-butanol extract, and aqueous phase extract. The ethyl acetate extract was separated into distillate(1)-(6)using D101 macroporous resin column chromatography. The experiment was divided into control group, LPS model group, L. nervosa extract group, and LPS + L. nervosa group. LPS was utilized to induce a neuroinflammatory cell model in BV-2 microglial cells. The Griess test was utilized for detecting the production of nitric oxide(NO) in the cell supernatant. Cell viability was detected by MTT assay. The release of interleukin-6(IL-6) and tumor necrosis factor alpha(TNF-α) in the cell supernatant was quantified using ELISA.RT-qPCR was utilized to assess the m RNA levels of pro-inflammatory cytokines inducible nitric oxide synthase(iNOS), cyclooxygenase-2(COX-2), interleukin( IL)-6, IL-1β, and TNF-α. The protein expression of i NOS, COX-2, nuclear factor kappa-B p65(p65), p-p65, extracellular signal-regulated kinase(ERK), p-ERK, c-jun N-terminal kinase(JNK), p-JNK, p38 mitogen-activated protein kinase(p38), and p-p38 MAPK(p-p38) were also evaluated by Western blot. The chemical composition of active substances in L. nervosa was analyzed using the UHPLC-Q-Exactive Orbitrap technology and literature comparison. Our findings indicate that extracts from different parts of L. nervosa exhibit a significant reduction in the release of NO from LPS-induced BV-2 microglial cells.Specifically, the ethyl acetate extract demonstrates the most notable inhibitory effect without causing cell toxicity. Additionally, the distillate(6) extracted from the ethyl acetate exhibits a reduction in the m RNA and protein levels of i NOS, COX-2, IL-6, IL-1β, and TNF-α in a dose-dependent manner, and it inhibits the protein expression of p-p65, p-ERK, p-p38, and p-JNK in LPS-induced BV-2 microglial cells. A total of 79 compounds in the distillate(6) were identified by mass spectrometry, including 12 confirmed compounds with anti-inflammatory effects. This study confirmed the remarkable efficacy of L. nervosa extract in the treatment of neuroinflammation, which may be achieved through the inhibition of NF-κB and MAPK signaling pathways.

Physicochemical characterization of the fraction components of the theabrownin isolates from Pu'er tea

This study focuses on the isolation of theabrownins (TBs) from Pu'er tea. This isolation process involves low temperature water extraction, membrane separation, and multi-stage extraction with organic solvents. Subsequently, purification was performed using column chromatography with macroporous adsorbent resin D-101 and ethanol aqueous solution with a concentration of 0–64.81 g/100g. Five purified fractions were obtained, namely TBs-D1, TBs-D2, TBs-D3, TBs-D4, and TBs-D5, collectively referred to as TBs-D(1–5). These fractions were found to contain TBs and polysaccharides but lacked catechins, caffeine, theaflavin, and free protein. The average molecular weights of these fraction components exceeded 104 Da. These results indicated that TBs-D(1–5) were not free, but combined with tea polysaccharide conjugates (TPC), that is, TBs-TPC complexes. Furthermore, ultraviolet–visible spectrophotometry and Flourier transformation infrared spectroscopy confirmed the nature of TBs-D(1–5) as TBs-TPC complexes. Fluorescence spectra revealed that polysaccharides exert a quenching effect on the intrinsic fluorescence of TBs. Atomic force microscopy indicated that the sample forms a spherical cluster due to the aggregation of theabrownin molecules. Additionally, measurements of interfacial tension and water contact angle demonstrated that TBs-D(1–5) exhibit some amphiphilic properties. This research provides a theoretical foundation for the industrial production and utilization of TBs.

Preparation, Purification, Characterization and Antioxidant Activity of Rice Bran Fermentation Broth with Hypsizigus marmoreus

The main purpose of this study was to investigate the composition, characterization and antioxidant activity of rice bran fermentation broth, and provide a new way for high-value utilization of rice bran. Firstly, we fermented rice bran with Hypsizigus marmoreus and purified fermentation broth with macroporous resins. We took feruloyl oligosaccharides (FOs) concentration as the measure index, and the results showed that the maximum concentration of FOs was 0.72 mmol/L on the 6th day of rice bran fermentation. We took D101 macroporous resin as adsorption resin for rice bran fermentation broth, and the result showed that FOs concentration reached 2.38 mmol/L with the optimal purification process at pH 4.5, temperature 29 °C, ethanol concentration 55%, sample flow rate 1.5 mL/min, sample concentration 1.7 mL/min and elution flow rate 2.0 mmol/L. Secondly, the characters of rice bran fermentation broth were identified by high-performance liquid chromatography (HPLC) and Fourier-transform infrared spectroscopy (FTIR). These methods showed the presence of ferulic acid (FA), arabinose, xylose and glucose in rice bran fermentation broth. Finally, the in vitro antioxidant activities of rice bran fermentation broth were tested and the result showed that fermentation broth had good antioxidant activities and significantly improved after purification.

Sasanquasaponin from Camellia oleifera Abel Exerts an Anti-Inflammatory Effect in RAW 264.7 Cells via Inhibition of the NF-κB/MAPK Signaling Pathways.

Sasanquasaponin (SQS), a secondary metabolite that is derived from Camellia seeds, reportedly possesses notable biological properties. However, the anti-inflammatory effects of SQS and its underlying mechanisms remain poorly explored. Herein, we aimed to investigate the anti-inflammatory properties of SQS against lipopolysaccharide (LPS)-induced inflammatory responses in RAW264.7 cells, focusing on the nuclear factor-κB (NF-κB) and MAPK signaling pathways. SQS was isolated using a deep eutectic solvent and D101 macroporous adsorption resin and analyzed using high-performance liquid chromatography. The viability of LPS-stimulated RAW264.7 was assessed using the CCK-8 assay. The presence of reactive oxygen species (ROS) was evaluated using 2',7'-dichlorofluorescein-diacetate. The expression levels of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6) were detected using reverse transcription-quantitative PCR and ELISA. Western blot was performed to analyze the protein expression of LPS-induced RAW264.7 cells. Herein, SQS exhibited anti-inflammatory activity: 30 μg/mL of SQS significantly reduced ROS generation, inhibited the LPS-induced expression of iNOS and COX-2, and attenuated the production of pro-inflammatory cytokines IL-1β, IL-6, and TNF-α. The anti-inflammatory activity was potentially mediated by inhibiting the phosphorylation of IκBα and p65 in the NF-κB signaling pathway and the phosphorylation of ERK and JNK in the MAPK signaling pathway. Accordingly, SQS could inhibit inflammation in LPS-induced RAW264.7 cells by suppressing the NF-κB and MAPK signaling pathways. This study demonstrated the potential application of SQS as an anti-inflammatory agent.

Chemical constituents from Alangium chinense subsp. pauciflorum

Chemical constituents of 70% ethanol extract of Alangium chinense subsp. pauciflorum were investigated. The 70% ethanol extract of A. chinense subsp. pauciflorum was isolated and purified by D-101 macroporous resins, silica gel, Sephadex LH-20 and other methods. As a result, nineteen compounds were isolated and identified as 4-cyclohexene-1α,2α,3α-triol-1-O-β-D-glucoside(1), 1β,4α,6α,13-tetrahydroxy-eudesm-11(12)-ene(2), sucrose(3), 1'-O-benzyl-α-L-rhamnopyranosyl-(1″→6')-β-D-glucopyranoside(4), bis(2-ethylhexyl)benzene-1,2-dicarboxylate(5),(Z)-10-heneicosenoic acid(6), di-O-methylcrenati(7), methyl-α-D-fructofuranoside(8), β-daucosterol(9), syringic acid(10), vanillicacid(11), octacosanol(12), isoarborinol(13), 2,7-dihydroxy-6-methyl-4-(1-methylethyl)-1-naphthalenecarboxylate(14),vanillin(15), coniferyl aldehyde(16), 9(11)-dehydroergosterolperoxide(17), 5α,8α-epidioxy-(22E,24R)-ergosta-6,22-dien-3β-ol(18), β-sitosterol(19), respectively. Compounds 1 and 2 were new compounds, compounds 5-11, 13, 15-18 were isolated from Alangium for the first time.The anti-inflammatory activity of compourd 1 was determinded by the LPS-induced RAW264.7 macrophage inflammation model. The results showed that the new compound 1 has a certain inhibitory effect on LPS-induced NO production of RAW264.7 cells, and the inhibitory rate was 54.57%.