Ilex Rotunda Research Articles

Quantification and Pharmacokinetics Study of Pedunculoside in Rats by Using UPLC-MS/MS

Background: Pedunculoside (PE) is a triterpene saponin from the barks of Ilex rotunda, a Traditional Chinese Medicine called Jiubiying, which is used for the treatment of cold and fever, tonsillitis, sore throat, acute and chronic hepatitis, etc. Previous studies have confirmed that crude extract orally has a significant therapeutic effect on myocardial infarction. Methods: A simple, sensitive, and specific method by using UPLC-MS/MS to study the pharmacokinetics of PE in rats was developed and validated, with ilexsaponin A as an internal standard. Methanol was used as a protein precipitation reagent for blood sample extraction. A Waters Acquity C18 column (2.1 mm × 50 mm, 1.7 μm) was used for chromatographic separation with a gradient elution of CH3CN: 0.1% formic acid (0.3 mL·min-1). Negative ion electrospray ionization was used for detection in multiple reaction monitoring mode. Results: PE was linear within the concentration range of 0.14-1118.00 ng/mL. The LLOQ was 0.14 ng/mL for the plasma samples. The intra-day and inter-day precision were ranged from 1.18% to 10.48%, while the accuracy ranged from -1.32% to 1.68%, indicating satisfactory precision and accuracy of the assay. The extraction recoveries for PE and IS were ranged from 81.40% to 86.65%, with no significant variation among the three concentrations, respectively. PE remained stable at room temperature (25°C) for 3 h, in auto-sampler (4°C) for 24 h, after three freeze-thaw cycles, and in long-term storage at ‒20°C for 30 days. The PK results of PE indicated its poor oral bioavailability (3.37%). Conclusion: Non-compartmental pharmacokinetics parameters indicated that PE was rapidly distributed to the tissues and metabolized. The pharmacokinetic data of this paper highlighted the first-time report of PE oral bioavailability with two different administration manners, which will help to better understand how PE metabolized in rats and exert its pharmacological effect in vivo.

Rotundic acid reduces LPS-induced acute lung injury in vitro and in vivo through regulating TLR4 dimer.

Acute lung injury (ALI) is a serious clinical disease. Rotundic acid (RA), a natural ingredient isolated from Ilex rotunda Thunb, exhibits multiple pharmacological activities. However, RA's therapeutic effect and mechanism on ALI remain to be elucidated. The present study aimed to further clarify its regulating effects on inflammation in vitro and in vivo. Our results indicated that RA significantly inhibited the overproduction of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), cyclooxygenase-2 (COX-2), and inducible nitric oxide synthase (iNOS). RA decreased ROS production and calcium influx. In addition, RA inhibited the activation of PI3K, MAPK, and NF-κB pathways and enhanced the activity of nuclear factor E2-related factor 2 (Nrf2) signaling. The cellular thermal shift assay and docking results indicated that RA bind to TLR4 to block TLR4 dimerization. Furthermore, RA pretreatment effectively inhibited ear edema induced by xylene and LPS-induced endotoxin death and had a protective effect on LPS-induced ALI. Our findings collectively indicated that RA has anti-inflammatory effects, which may serve as a potential therapeutic option for pulmonary inflammation.

Absolute bioavailability, dose proportionality, and tissue distribution of rotundic acid in rats based on validated LC-QqQ-MS/MS method

Rotundic acid (RA), an ursane-type pentacyclic triterpene acid isolated from the dried barks of Ilex rotunda Thunb. (Aquifoliaceae), possesses diverse bioactivities. To further study its pharmacokinetics, a simple and sensitive liquid chromatography with triple quadrupole mass spectrometry (LC-QqQ-MS/MS) method was developed and validated to quantify RA concentration in rat plasma and tissue using etofesalamide as an internal standard (IS). Plasma and tissue samples were subjected to one-step protein precipitation. Chromatographic separation was achieved on a ZORBAX Eclipse XDB-C18 column (4.6 mm × 50 mm, 5 μm) under gradient conditions with eluents of methanol:acetonitrile (1:1, V/V) and 5 mM ammonium formate:methanol (9:1, V/V) at 0.5 mL/min. Multiple reaction monitoring transitions were performed at m/z 487.30 → 437.30 for RA and m/z 256.10 → 227.10 for IS in the negative mode. The developed LC-QqQ-MS/MS method exhibited good linearity (2–500 ng/mL) and was fully validated in accordance with U.S. Food and Drug Administration bioanalytical guidelines. Dose proportionality and bioavailability in rats were determined by comparing pharmacokinetic data after single oral (10, 20, and 40 mg/kg) and intravenous (10 mg/kg) administration of RA. Tissue distribution was studied following oral administration at 20 mg/kg. The results showed that the absolute bioavailability of RA after administration at different doses ranged from 16.1% to 19.4%. RA showed good dose proportionality over a dose range of 10–40 mg/kg. RA was rapidly absorbed in a dose-dependent manner and highly distributed in the liver. In conclusion, this study is the first to systematically elucidate the absorption and distribution characteristics of RA in rats, which can provide additional information for further development and evaluation of RA in drug metabolism and pharmacokinetic studies.

Comparative analysis and phylogenetic investigation of Hong Kong Ilex chloroplast genomes

Ilex is a monogeneric plant group (containing approximately 600 species) in the Aquifoliaceae family and one of the most commonly used medicinal herbs. However, its taxonomy and phylogenetic relationships at the species level are debatable. Herein, we obtained the complete chloroplast genomes of all 19 Ilex types that are native to Hong Kong. The genomes are conserved in structure, gene content and arrangement. The chloroplast genomes range in size from 157,119 bp in Ilex graciliflora to 158,020 bp in Ilex kwangtungensis. All these genomes contain 125 genes, of which 88 are protein-coding and 37 are tRNA genes. Four highly varied sequences (rps16-trnQ, rpl32-trnL, ndhD-psaC and ycf1) were found. The number of repeats in the Ilex genomes is mostly conserved, but the number of repeating motifs varies. The phylogenetic relationship among the 19 Ilex genomes, together with eight other available genomes in other studies, was investigated. Most of the species could be correctly assigned to the section or even series level, consistent with previous taxonomy, except Ilex rotunda var. microcarpa, Ilex asprella var. tapuensis and Ilex chapaensis. These species were reclassified; I. rotunda was placed in the section Micrococca, while the other two were grouped with the section Pseudoaquifolium. These studies provide a better understanding of Ilex phylogeny and refine its classification.

Neighborhood effects and environmental variables drive sapling growth in a young subtropical tree plantation

We established a forest biodiversity and ecosystem functioning experiment in southern China by planting 20,480 saplings of eight subtropical tree species in 320 plots in monocultures and mixtures of two, four and eight species to explore the biodiversity-ecosystem functioning (BEF) relationships and the underlying coexistence determinants. In this study, we aimed to report the growth of these planted saplings and explore its abiotic and biotic driving factors. Specifically, we analyzed the relative importance of sapling species identity, initial size, neighborhood interactions, neighborhood diversity, and environmental factors to sapling growth in the early stage of our BEF experiment. We found that the species of Ilex rotunda, Pinus massoniana and Elaeocarpus sylvestris grew much faster than other five species, followed by Castanopsis carlesii, Cinnamomum camphora and Schima superba. Erythrophleum fordii and Castanopsis fissa had the lowest growth rates in ground diameter and height. In addition, the relative growth rates of saplings were positively related to the neighborhood interactions, neighborhood diversity and soil nutrients, but were negatively related to initial size. Our study stresses the importance of better understanding the effects of environmental and local neighborhood conditions on initial growth of subtropical forest tree species in a mixed plantation, and provides reference for other BEF experiments in China and elsewhere.

Evaluation of the efficiency of the imperative ecological remediation regarding tree growth, root development, and edaphic properties after Typhoon Hato (2017) in Zhuhai, China.

Background: urban forest in coastal cities encounters multiple disturbances of frequent typhoon events caused by global change, under which ecological remediation can help to improve urban environment. We measured and analyzed the growth and ecosystem services of four newly-planted tree species in Zhuhai after Typhoon Hato (2017), aiming to evaluate the efficiency of the ecological remediation. Methods: National Meteorological Information Center of China supplied climate variables. From June 2018 to December 2019, we measured soil physical and chemical properties, above- and below-ground development regarding stem, tree height, and root growth of all the selected tree species. Results: Sl (Sterculia lanceolata Cav.), Ir (Ilex rotunda Thunb), Ss (Schima superba Gardn. et Champ.) could be more wind-resistant from the above-ground morphological perspective. For the below-ground process, Sl was the only tree species with continuous development, while Ir, Ss, and Es (Elaeocarpus sylvestris (Lour.) Poir.) decreased. Furthermore, Sl, Ir, and Ss maintained their investment in deep roots when Es had apparent deep root biomass reduction. The edaphic condition showed notable improvement in chemical properties rather than physical properties, especially for AN (available nitrogen), AK (available potassium), and SOM (soil organic matter). Conclusions: The ecological remediation in Zhuhai after Typhoon Hato (2017) was efficient, and in the future, tree species like Sl with advantages in root development and morphological profile were preferentially recommender for plantation in typhoon-affected areas.

Structural characterisation and screening of triterpene saponins in the bark of Ilex rotunda using high-performance liquid chromatography coupled to electrospray ionisation and quadrupole time-of-flight mass spectrometry.

The bark of Ilex rotunda (BIr) has been used as a traditional Chinese medicine for a long time in China. Triterpene saponins (TSs) were considered to be the major bioactive ingredients and possessed antiplatelet aggregation activity. Identification of TSs in BIr is a necessary step to understand the biological properties. This article describes a method for the rapid identification of TSs in BIr. An analytical method based on high-performance liquid chromatography coupled to electrospray ionisation and quadrupole time-of-flight tandem mass spectrometry (HPLC-ESI-QTOF-MS/MS) was established to characterise the TSs in BIr. Chromatographic separation was performed on an Agilent poroshell 120 EC-C18 column with gradient elution using water containing 0.1% (v/v) formic acid and acetonitrile as the mobile phase. A total of 26 TSs were identified or tentatively identified from BIr. Of them, 23 TSs were first reported from BIr. This was the first comprehensive investigation on TSs in BIr, and the results formed the foundation for the further development and utilisation of BIr in the future.

Extraction and purification of pedunculoside from the dried barks of Ilex rotunda using crystallization combined with polyamide column chromatography

ABSTRACT An efficient and low-cost method was developed for the preparation of pedunculoside from the dried barks of Ilex rotunda using crystallization combined with polyamide column chromatography. The kinetics of ethanol extraction was first evaluated and then the extract was concentrated to remove ethanol. Pedunculoside was partially crystallized after ethanol removal and those left in solution were further separated by polyamide column chromatography. To optimize the operating parameters for chromatography, the static and dynamic adsorption/desorption experiments were conducted. The purity of pedunculoside in the product increased to 85.69%, with an overall yield of 66.71%. The extraction and purification method developed herein provides a reference standard and can be used as the starting material for further studies.

Large-Scale Screening of 239 Traditional Chinese Medicinal Plant Extracts for Their Antibacterial Activities against Multidrug-Resistant Staphylococcus aureus and Cytotoxic Activities.

Novel alternative antibacterial compounds have been persistently explored from plants as natural sources to overcome antibiotic resistance leading to serious foodborne bacterial illnesses. In this study, the ethanolic extracts from 239 traditional Chinese medicinal plants (TCMP)’ materials were screened to discover promising candidates that have strong antibacterial properties against multidrug-resistant Staphylococcus (S.) aureus and low cytotoxicity. The results revealed that 74 extracts exhibited good antibacterial activities (diameter of inhibition zone (DIZ) ≥ 15 mm). Furthermore, 18 extracts (DIZ ≥ 20 mm) were determined their minimum inhibitory concentrations (MIC) and minimum bactericide concentrations (MBC), ranging from 0.1 to 12.5 mg/mL and 0.78 to 25 mg/mL, respectively. In addition, most of the 18 extracts showed relatively low cytotoxicity (a median lethal concentration (LC50) >100 µg/mL). The 18 extracts were further determined to estimate possible correlation of their phenolic contents with antibacterial activity, and the results did not show any significant correlation. In conclusion, this study selected out some promising antibacterial TCMP extracts with low cytotoxicity, including Rhus chinensis Mill., Ilex rotunda Thunb., Leontice kiangnanensis P.L.Chiu, Oroxylum indicum Vent., Isatis tinctorial L., Terminalia chebula Retz., Acacia catechu (L.f.) Willd., Spatholobus suberectus Dunn, Rabdosia rubescens (Hemsl.) H.Hara, Salvia miltiorrhiza Bunge, Fraxinus fallax Lingelsh, Coptis chinensis Franch., Agrimonia Pilosa Ledeb., and Phellodendron chinense C.K.Schneid.

Rotundic Acid Protects against Metabolic Disturbance and Improves Gut Microbiota in Type 2 Diabetes Rats.

Rotundic acid (RA) is a major triterpene constituent in the barks of Ilex rotunda Thunb, which have been widely used to make herbal tea for health care in southern China. RA has a variety of bioactivities such as anti-inflammation and lipid-lowering effect. However, little is known about the effects and mechanisms of RA on metabolic disturbance in type 2 diabetes (T2D) and its effect on gut microbiota. A T2D rat model induced by high fat diet (HFD) feeding and low-dose streptozotocin (STZ) injection was employed and RA showed multipronged effects on T2D and its complications, including improving glucolipid metabolism, lowering blood pressure, protecting against cardiovascular and hepatorenal injuries, and alleviating oxidative stress and inflammation. Furthermore, 16s rRNA gene sequencing was carried out on an Illumina HiSeq 2500 platform and RA treatment could restore the gut microbial dysbiosis in T2D rats to a certain extent. RA treatment significantly enhanced the richness and diversity of gut microbiota. At the genus level, beneficial or commensal bacteria Prevotella, Ruminococcus, Leuconostoc and Streptococcus were significantly increased by RA treatment, while RA-treated rats had a lower abundance of opportunistic pathogen Klebsiella and Proteus. Spearman’s correlation analysis showed that the abundances of these bacteria were strongly correlated with various biochemical parameters, suggesting that the improvement of gut microbiota might help to prevent or attenuate T2D and its complication. In conclusion, our findings support RA as a nutraceutical agent or plant foods rich in this compound might be helpful for the alleviation of T2D and its complications through improving gut microbiota.

Pedunculoside attenuates pathological phenotypes of fibroblast-like synoviocytes and protects against collagen-induced arthritis

Objectives: The discovery of alternative and well-tolerated anti-arthritic drugs, especially from natural products, is becoming an area of active research. Pedunculoside (PE) is a novel triterpene saponin extracted from the dried bark of Ilex rotunda Thunb. Limited published papers have reported its pharmacological properties, including anti-inflammatory, anti-myocardial ischaemia, anti-liver injury, and hypocholesterolaemic activities. However, the effect of PE on rheumatoid arthritis (RA) remains unknown. Here, we investigated the anti-arthritic effect of PE in both in vitro and in vivo models.Method: The inhibitory effects of PE on proliferation, migration, and production of inflammatory mediators in primary fibroblast-like synoviocytes (FLSs) were examined by a 5-ethynyl-2ʹ-deoxyuridine incorporation assay, wound-healing assay, and real-time polymerase chain reaction, respectively. Cellular signalling mechanisms were analysed by Western blot. The in vivo studies were performed using a collagen-induced arthritis (CIA) rat model. Multiple methods, including arthritis scoring, enzyme-linked immunoassay, radiography, and histopathological assessment, were used to evaluate the therapeutic effects of PE on CIA rats.Results: The in vitro studies revealed that PE significantly inhibited proliferation and migration of FLSs. PE also decreased the production of pro-inflammatory cytokines, including interleukin-1β (IL-1β), IL-6, IL-8, and tumour necrosis factor-α (TNF-α). Western blot results suggested that PE suppressed TNF-α-stimulated activation of p38 and extracellular signal-regulated kinase. The in vivo studies showed that PE treatment significantly inhibited synovial inflammation and bone destruction in CIA rats.Conclusion: These results demonstrate that PE exerts an inhibitory role in FLSs and CIA rats, and therefore may have therapeutic value for the treatment of RA.

The first definite record of Sinomegoura citricola (van der Goot, 1917) (Hemiptera: Aphididae) occurring on Ilex rotunda (Aquifoliaceae)

In April 2018, the occurrence of Sinomegoura citricola was recorded on Ilex rotunda (Aquifoliaceae) in Japan. Both aphid identification with mounted specimens and rearing experiments with the plant shoots revealed that I. rotunda is a definite host for S. citricola. This is believed to be the first study to report a species of the family Aquifoliaceae as a certain host plant of S. citricola, emphasizing that the aphid is a polyphagous species.

Extract of Ilex rotunda Thunb alleviates experimental colitis-associated cancer via suppressing inflammation-induced miR-31-5p/YAP overexpression

BackgroundIlex rotunda Thunb is a traditional medicine used in China treating colitis clinically. Triterpenoids is one of its main components. However, the detailed pharmacological activity and the component responsible for its clinical effects are still elusive. PurposeTo test the in vivo colitis-associated cancer (CAC) preventive effect of the water fraction extracted from the roots of I. rotunda, and to evaluate its microRNA (miRNA)-related mechanism. Study design and methodsMale or female C57BL/6 mice (12 weeks of age) were used to construct the azoxymethane (AOM)/dextran sulfate sodium (DSS)-induced CAC. 12.5 mg/kg and 25.0 mg/kg of the standardized water extract of I. rotunda (WIR), being equal to 4.29 and 8.58 g of the raw medicine respectively, were adopted to treat the AOM/DSS-induced CAC from the fourth week and continued for 5 weeks. Mice were killed two weeks after the end of the last round of DSS by cervical dislocation. ResultsThe chemical analysis of WIR revealed the presence of 21 compounds. The syringing and caffeic acid (1-hydroxyl-4-O-β-D-glucopyranosylprenyl)-ester are the main components of WIR, counting for 8.27% and 5.71% of the water extract respectively. The levels of miR-31-5p were up-regulated in both thp1 and Caco2 cells (p < 0.05) stimulated by either IL-6 or TNF-α, and WIR could restore miR-31-5p levels in the IL-6/TNF-α-stimulated thp-1 and Caco2 cells. Furthermore, WIR decreased TNF-α and IL-6 levels in PMA-differentiated thp-1 cells stimulated by LPS via NF-κB pathway (p < 0.05), suggesting that WIR could restore miR-31-5p expression via down-regulating IL-6 and TNF-α levels. In vivo study showed that oral administration of WIR (25 mg/kg) produced a significant inhibition on the atypical hyperplasia, as well as the release and the expression of IL-6 and TNF-α in the colon tissue. The in vivo transcription of other pro-inflammatory mediators such as iNOS, IL-11, and IL-17A were also attenuated by WIR administration (25 mg/kg, p < 0.05). Meanwhile, WIR (25 mg/kg) restored the miR-31-5p level which was up-regulated in the CAC model group, and ectopic expressions of the miR-31-5p down-stream LATS2 and YAP genes in the hippo pathway were also modulated by the WIR (25 mg/kg) treatment. ConclusionThe present study suggests that WIR exerts intestinal anti-inflammatory and CAC preventive effects in an experimental CAC mouse model. The CAC preventive effect can be attributed to the suppression of hippo pathway activated by the inflammatory cytokines, indicating that WIR can be potentially used as an herbal product for CAC prevention. Therefore, there is an emergent need for further evaluation of the main components in WIR to determine the definite bioactive component responsible for the CAC preventive activity.

Natural CAC chemopreventive agents from Ilex rotunda Thunb.

Colitis-associated cancer (CAC) is one of the most serious complications of inflammatory bowel disease. The pathogenesis of CAC is complicated and so far elusive, and the anti-inflammatory effect does not assure CAC preventive activity, making it difficult to discover CAC preventive drugs. In this study, we report the CAC preventive effect of the ethyl acetate (EIR) of Ilex rotunda Thunb., a traditional Chinese herbal medicine being clinically used to treat intestinal disease. We also report the results of screening for CAC preventive agents from EIR via a nuclear factor-kappa B (NF-κB) translocation model in Caco2 cells, since activated NF-κB can be used by tumor cells at the early stage of tumorigenesis. Twenty-four components were isolated from EIR and identified by multiple chromatography and spectral analysis. MTT experiments in IEC-6 and RAW264.7 cells showed that all 24 compounds were toxic-free to normal cell lines. Furthermore, compound rotundic acid (RA) (19) exhibited an inhibitory effect on LPS-induced NF-κB translocation in Caco2 cells. Moreover, RA did not induce apoptosis in Caco2 tumor cells while possessing an anti-inflammatory effect both in immune and intestinal epithelium cells (RAW264.7 and IEC-6 cells, respectively). Removing RA (19) and its 28-O-glucopyranoside (17) from EIR definitely undermined the in vivo CAC preventive activity of EIR. Therefore, the current study suggested that RA (19) could be a potential therapeutic agent against CAC.

A facile and selective approach to the qualitative and quantitative analysis of triterpenoids and phenylpropanoids by UPLC/Q-TOF-MS/MS for the quality control of Ilex rotunda

Ilex rotunda, in which triterpenoids and phenylpropanoids are major bioactive constituents, has been widely used in traditional Chinese medicines. In this study, a validated UPLC/Q-TOF-MS/MS method was developed to simultaneously identify and quantify the triterpenoids and phenylpropanoids in the stem bark, fruit, leaves, roots and stem xylem of this herbal medicine. A total of seventy triterpenoids and twelve phenylpropanoids were identified with the assistance of the modified mass defect filter and key product ion filter data processing strategies, and forty-eight of them were confirmed by reference substances. Meanwhile, the contents of twelve triterpenoids and three phenylpropanoids in the five plant parts were determined with good linearity (R2 ≥ 0.9993), precision (RSD ≤ 2.04%), repeatability (RSD ≤ 1.99%), stability (RSD ≤ 1.88%) and recovery (96.65–103.17% and RSD ≤ 3.54%). Furthermore, PCA and OPLS-DA methods were employed to visualize the relationships and discrimination of the forty-two stem bark samples from two origins based on the contents of fifteen analytes. Our findings may provide early scientific evidence for quality control and for elucidating the therapeutic principle of Ilex rotunda.

Pedunculoside, a novel triterpene saponin extracted from Ilex rotunda, ameliorates high-fat diet induced hyperlipidemia in rats

Pedunculoside (PE) is a novel triterpene saponin extracted from the dried barks of Ilex rotunda Thunb. The present study aims to explore lipid-lowering effects of PE on hyperlipidemia rat induced by high-fat diet. The rats were fed with the high-fat diet and subjected to intragastric administration of PE at doses of 30, 15, or 5 mg/kg daily for 7 weeks. The results demonstrated that treatment with PE for 7-week dramatically decreased serum total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) and reduced liver TC in hyperlipidemia rat induced by high-fat diet. Furthermore, the results also showed that PE modulated the expression of enzymes involved in lipid metabolism including peroxisome proliferator-activated receptor α (PPAR-α), sterol regulatory element-binding protein 1 (SREBP-1), fatty acid synthase (FAS) and stearoyl CoA desaturase-1 (SCD-1) mRNA in liver. Besides, PE-treated group decreased weights and diameters of epididymal adipose hyperlipidemia rat. Mechanism study demonstrated that PE regulated PPAR-γ, CCAAT/Enhancer-binding Protein α (C/EBPα)、and SREBP-1 expression as well as inhibited phosphorylation of AMPK in MDI (methylisobutylxanthine, dexamethasone, insulin) induced-3T3L1 cells. Molecular Docking confirmed interaction between PE with proteins involving PPAR-γ, C/EBPα and SREBP-1. In summary, these findings may support that PE is a novel lipid-lowering drug candidate.

Triterpenoids with antiplatelet aggregation activity from Ilex rotunda

Phytochemical studies on the barks of Ilex rotunda Thunb. had resulted in the isolation of seven previously undescribed triterpenoids, rotundinosides E-K, along with sixteen known ones. The structures of previously undescribed compounds were elucidated on the basis of extensive spectroscopic analysis and the sugar moieties were further identified by HPLC and GC after acid hydrolysis. Among the isolates, rotundinoside F featured a rare triterpene-phenylpropanoid hybrid structure and rotundinoside H was an uncommon triterpene saponin with α-linked glucopyranosyl moiety at C-3. The antiplatelet aggregation of all compounds were evaluated against ADP induced rat platelet aggregation in vitro, and five compounds exhibited moderate inhibitory effects with IC50 values ranging from 22.4 to 32.8 μM.

A simple method for extraction and purification of pedunculoside from the dried barks of Ilex rotunda and its inhibitory effect on pancreatic lipase in vitro

ABSTRACTA simple and efficient method was developed for the preparation of pedunculoside from the dried barks of Ilex rotunda. Pedunculoside was extracted by heat reflux. The optimal conditions were determined by the response surface methodology (40% ethanol concentration, 14 mL/g solvent/material loading level and 90 min extraction time with the dried barks being extracted twice). After extraction and condensing the crude extract, pedunculoside was directly purified through crystallization in water with the addition of ethyl acetate. Pedunculoside (96.9% purity) was obtained with 48.2% recovery after the extraction and purification. Compared with the reported chromatographic methods, this strategy is simple, eco-friendly, and economical. The purified pedunculoside showed an inhibitory effect on pancreatic lipase in vitro (IC50 = 80.8 μg/mL), suggesting its anti-obesity potential.

Resveratrol-Mg hydroxide complex display enhanced bioavailability: A possible application in DM-1 disease

Phytochemical studies on the barks of Ilex rotunda Thunb. had resulted in the isolation of seven previously undescribed triterpenoids, rotundinosides E-K, along with sixteen known ones. The structures of previously undescribed compounds were elucidated on the basis of extensive spectroscopic analysis and the sugar moieties were further identified by HPLC and GC after acid hydrolysis. Among the isolates, rotundinoside F featured a rare triterpene-phenylpropanoid hybrid structure and rotundinoside H was an uncommon triterpene saponin with α-linked glucopyranosyl moiety at C-3. The antiplatelet aggregation of all compounds were evaluated against ADP induced rat platelet aggregation in vitro, and five compounds exhibited moderate inhibitory effects with IC50 values ranging from 22.4 to 32.8 μM.

Fluorescence Response of Greening Seedling of Four Tree Species to Low Temperature Stress

Fluorescence indexes of Cassia bakeriana, Syzygium hainanense, Ilex rotunda and Phoebe sheareri seedlings were determined under simulated low temperature stress using artificial climate chambers. The results showed that with increasing low temperature time, the minimal initial fluorescence Fo of C. bakeriana fluctuated, S. hainanense and P. sheareri decreased followed by an increase, I. rotunda decreased and then fluctuated; The biggest fluorescent Fm of C. bakeriana and I. rotunda increased followed by a decrease and increased again, S. hainanense continuously decreased; the largest PSII photochemical efficiency Fv/Fm of C. bakeriana and I. rotunda S. hainanense continuously decreased, S. hainanense increased followed by a decrease, whereas P. sheareri decreased followed by an increase; the PSII actual light quantum yield Y(II) and photosynthetic electron transport rate ETR of seedlings of the four tree species decreased; non‐photochemical quenching of C. bakeriana and S. hainanense fluctuated, whereas that of I. rotunda and P. sheareri continuously decreased. Two days after low‐temperature release, Fo of seedlings of the four tree species decreased, Fm of C. bakeriana, S. hainanense and I. rotunda, and Fv/Fm, Y(II), ETR and NPQ of seedlings of the four tree species increased. The fluorescence parameters of seedlings of the four tree species was evaluated using principal component analysis, indicating that cold‐resistance ability decreased in the order of C. bakeriana > P. sheareri > I. rotunda > S. hainanense