Plant-derived monoterpene S-linalool and ß-ocimene generated by CsLIS and CsOCS-SCZ are key chemical cues for attracting parasitoid wasps for suppressing Ectropis obliqua infestation in Camellia sinensis L.

Insect-induced plant volatile organic compounds (VOCs) may function as either direct defence molecules to deter insects or indirect defence signals to attract the natural enemies of the invading insects. Tea (Camellia sinensis L.), an important leaf-based beverage crop, is mainly infested by Ectropis obliqua which causes the most serious damage. Here, we report a mechanistic investigation of tea plant-derived VOCs in an indirect defence mechanism against E. obliqua. Parasitoid wasp Parapanteles hyposidrae, a natural enemy of E. obliqua, showed strong electrophysiological response and selection behaviour towards S-linalool and ß-ocimene, two monoterpenes with elevated emission from E. obliqua-damaged tea plants. Larvae frass of E. obliqua, which also released S-linalool and ß-ocimene, was found to attract both mated female or male Pa. hyposidrae according to gas chromatography-electroantennogram detection and Y-tube olfactometer assays. In a field setting, both S-linalool and ß-ocimene were effective in recruiting both female and male Pa. hyposidrae wasps. To understand the molecular mechanism of monoterpenes-mediated indirect defence in tea plants, two novel monoterpene synthase genes, CsLIS and CsOCS-SCZ, involved in the biosynthesis of S-linalool or ß-ocimene, respectively, were identified and biochemically characterised. When the expression of these two genes in tea plants was inhibited by antisense oligodeoxynucleotide, both volatile emission and attraction of wasps were reduced. Furthermore, gene expression analysis suggested that the expression of CsLIS and CsOCS-SCZ is regulated by the jasmonic acid signalling pathway in the tea plant.

High-efficiency green management of potato late blight by a self-assembled multicomponent nano-bioprotectant.

Potato late blight caused by Phytophthora infestans is a devastating disease worldwide. Unlike other plant pathogens, double-stranded RNA (dsRNA) is poorly taken up by P. infestans, which is a key obstacle in using dsRNA for disease control. Here, a self-assembled multicomponent nano-bioprotectant for potato late blight management is designed based on dsRNA and a plant elicitor. Nanotechnology overcomes the dsRNA delivery bottleneck for P. infestans and extends the RNAi protective window. The protective effect of nano-enabled dsRNA against infection arises from a synergistic mechanism that bolsters the stability of dsRNA and optimizes its effective intracellular delivery. Additionally, the nano-enabled elicitor enhances endocytosis and amplifies the systemic defense response of the plants. Co-delivery of dsRNA and an elicitor provides a protective effect via the two aspects of pathogen inhibition and elevated plant defense mechanisms. The multicomponent nano-bioprotectant exhibits superior control efficacy compared to a commercial synthetic pesticide in field conditions. This work proposes an eco-friendly strategy to manage devastating plant diseases and pests.

Guiding ablation strategies for ventricular tachycardia in patients with structural heart disease by analyzing links and conversion patterns of traceable abnormal late potential zone.

BACKGROUND: Substrate-based ablation can treat uninducible or hemodynamically instability scar-related ventricular tachycardia (VT). However, whether a correlation exists between the critical VT isthmus and late activation zone (LAZ) during sinus rhythm (SR) is unknown. OBJECTIVE: To demonstrate the structural and functional properties of abnormal substrates and analyze the link between the VT circuit and abnormal activity during SR. METHODS: Thirty-six patients with scar-related VT (age, 50.0 ± 13.7 years and 86.1% men) who underwent VT ablation were reviewed. The automatic rhythmia ultrahigh resolution mapping system was used for electroanatomic substrate mapping. The clinical characteristics and mapping findings, particularly the LAZ characteristics during SR and VT, were analyzed. To determine the association between the LAZ during the SR and VT circuits, the LAZ was defined as five activation patterns: entrance, exit, core, blind alley, and conduction barrier. RESULTS: Forty-five VTs were induced in 36 patients, 91.1% of which were monomorphic. The LAZ of all patients was mapped during the SR and VT circuits, and the consistency of the anatomical locations of the LAZ and VT circuits was analyzed. Using the ultrahigh resolution mapping system, interconversion patterns, including the bridge, T, puzzle, maze, and multilayer types, were identified. VT ablation enabled precise ablation of abnormal late potential conduction channels. CONCLUSION: Five interconversion patterns of the LAZ during the SR and VT circuits were summarized. These findings may help formulate more precise substrate-based ablation strategies for scar-related VT and shorter procedure times.

Secondary Metabolites from the Cultures of Medicinal Mushroom Vanderbylia robiniophila and Their Tyrosinase Inhibitory Activities.

Vanderbylia robiniophila (Huaier in Chinese) has been used as a traditional herbal medicine in China for over 1600 years. However, the secondary metabolites of V. robiniophila have not been systematically examined. Corresponding chemical investigation in this study led to the discovery of two new compounds, (22E, 24R)-6ß, 7α-dimethoxyergosta-8(14), 22-diene-3ß, 5α-diol (1) and vanderbyliolide A (8), along with eight known ones (2-7, 9-10). Their structures were determined by extensive spectroscopic analyses and electronic circular dichroism (ECD) calculations. The tyrosinase inhibitory activity of all isolated compounds was evaluated, and compound 10 showed a potential tyrosinase inhibitory effect with an IC50 value of 60.47 ± 2.63 µM. Kinetic studies of the inhibition reactions suggested that 10 provides the inhibitory ability on tyrosinase in an uncompetitive way.

Optimization of 4-arylthiophene-3-carboxylic acid derivatives as inhibitors of ANO1: Lead optimization studies toward their analgesic efficacy for inflammatory pain.

Current pain management is largely limited to opioids and non-steroidal anti-inflammatory drugs. Developing new analgesic drugs remains important to address the unmet medical needs of chronic pain patients. Calcium-activated chloride channel anoctamin-1 (ANO1) is a potential analgesic target. ANO1 is activated by noxious stimuli in peripheral sensory neurons and further induced neural depolarization. Downregulation of ANO1 reduced hyperalgesia and allodynia caused by inflammation and nerve injury. Here we developed a series of 4-arylthiophene-3-carboxylic acid derivatives for proof-of-concept studies of ANO1-targeted analgesia. These efforts led to the identification of the compound DFBTA, 4-(4-chlorophenyl)-2-(2,5-difluorobenzamido)thiophene-3-carboxylic acid, which displays dramatic ANO1 inhibition with IC50 of 24 nM. DFBTA displays very weak cytotoxicity, cardiotoxicity, and acute toxicity (HEK293 proliferation IC50 > 30 µM, hERG IC50 > 30 µM, mouse minimum lethal dosage, MLD>1000 mg/kg), as well as excellent pharmacokinetics properties with oral bioavailability >75% and little brain penetration (<1.5% brain/plasma). Finally, the analgesic efficacy of ANO1 inhibitor was evaluated in animal models. DFBTA shown comparable efficacy to clinical drugs in all inflammatory pain models induced by complete Freund's adjuvant, formalin, and capsaicin. These works provide a useful tool compound and promising results for ANO1-targenting analgesic development.

Ultrasonic-assisted extraction of polyphenolic compounds from Paederia scandens (Lour.) Merr. Using deep eutectic solvent: optimization, identification, and comparison with traditional methods.

Ultrasonic-assisted extraction (UAE) coupled with deep eutectic solvent (DES) is a novel, efficient and green extraction method for phytochemicals. In this study, the effects of 16 DESs coupled with UAE on the extraction rate of polyphenols from Paederia scandens (Lour.) Merr. (P. scandens), an edible and medicinal herb, were investigated. DES synthesised with choline chloride and ethylene glycol at a 1:2 M ratio resulted in the highest extractability. Moreover, the effects of extraction parameters were investigated by using a two-level factorial experiment followed by response surface methodology The optimal parameters (water content in DES of 49.2%, the actual ultrasonic power of 72.4 W, and ultrasonic time of 9.7 min) resulted in the optimal total flavonoid content (TFC) (27.04 mg CE/g DW), ferric-reducing antioxidant power (FRAP) value (373.27 µmol Fe(Ⅱ)E/g DW) and 2,2'-azino-bis(3-ethylbenzthiazoline)-6-sulfonic acid radical (ABTS+) value (48.64 µmol TE/g DW), closely matching the experimental results. Furthermore, a comparison study demonstrated that DES-UAE afforded the higher TFC and FRAP value than traditional extraction methods. 36 individual polyphenolic compounds were identified and quantified by ultra-high-performance liquid chromatography-mass spectrometry (UHPLC-MS) in P. scandens extracts, and of which 30 were found in the extracts obtained by DES-UAE. Additionally, DES-UAE afforded the highest sum of individual polyphenolic compound content. These results revealed that DES-UAE enhanced the extraction efficiency for polyphenols and provided a scientific basis for further processing and utilization of P. scandens.

Effects of different extraction methods on contents, profiles, and antioxidant abilities of free and bound phenolics of Sargassum polycystum from the South China Sea.

Total phenolic content (TPC), phenolic profiles, and antioxidant activity of free and bound extracts of Sargassum polycystum, obtained by different extraction solvents and hydrolysis methods, were investigated. Aqueous acetone afforded the highest free TPC and antioxidant ability, followed by aqueous ethanol and aqueous methanol. Twelve free phenolic compounds were identified by ultra-high-performance liquid chromatography-mass spectrometry (UHPLC-MS), including two hydroxycinnamic acids, seven flavonoids, one stilbene, and two phlorotannins. Three to nine different free phenolic compounds were extracted by these solvents with different compositions, including nine by 70% acetone and eight by 70% methanol, 70% ethanol, and 50% ethanol. The highest total content of free phenolic compounds determined by high-performance liquid chromatography-diode array detection was obtained from 70% ethanol. Alkaline hydrolysis afforded higher bound TPC (274.27 mg GAE/100 g DW) and antioxidant ability than acid hydrolysis. Five bound phenolic compounds were characterized by UHPLC-MS and five were released from alkaline hydrolysis, whereas two were released from acid hydrolysis. Total content of bound phenolic compounds released by alkaline hydrolysis was 14.68-fold higher than that by acid hydrolysis. The free and bound TPC, phenolic profiles, and antioxidant activities depended on the extraction solvent used. These results indicate that S. polycystum is a potentially useful antioxidant source and contribute to the development of seaweed-based functional foods. PRACTICAL APPLICATION: Phenolics are usually divided into free and bound forms based on their extractability and interaction with cell wall components. The nutritional effects of bound phenolics in algae have long been neglected. These topics contribute to the development of seaweed-based functional foods.

Protective Effects of Astilbin Against Cadmium-Induced Apoptosis in Chicken Kidneys via Endoplasmic Reticulum Stress Signaling Pathway.

Cadmium (Cd) can cause endoplasmic reticulum stress (ERS) and apoptosis in animals. The kidney is an organ seriously affected by Cd because it can accumulate metal ions. Astilbin (ASB) is a dihydroflavonol rhamnoside, which has an anti-renal injury effect. This study aimed to evaluate the protective effect of ASB on Cd-induced ERS and apoptosis in the chicken kidney. In this study, a total of 120 1-day-old chickens were randomly divided into 4 groups. Chickens were fed with a basic diet (Con group), ASB 40 mg/kg (ASB group), CdCl2 150 mg/kg + ASB 40 mg/kg (ASB/Cd group), and CdCl2 150 mg/kg (Cd group) for 90 days. The results showed that Cd exposure induced pathological and ultrastructural damages and apoptosis in chicken kidneys. Compared with the Con group, metallothionein (MT1/MT2) level, nitric oxide (NO) content, inducible nitric oxide synthase (iNOS) activity, ERS-related genes 78-kDa glucose-regulated protein (Grp78), protein kinase PKR-like endoplasmic reticulum kinase (Perk), activating transcription factor 4 (Atf4) and CAAT/enhancer-binding protein (C/EBP) homologous protein (Chop), and pro-apoptotic gene B-cell lymphoma 2 (Bcl-2)-associated X (Bax), caspase-12, caspase-9, caspase-3 expression levels, and apoptotic rate were significantly increased in the Cd group. The expression level of Bcl-2 was significantly decreased in the Cd group. ASB/Cd combined treatment significantly improves the damage of chicken kidneys by ameliorating Cd-induced kidney ERS and apoptosis. Cd can cause the disorder of the GRP78 signal axis, activate the PERK-ATF4-CHOP pathway, aggravate the structural damage and dysfunction of ER, and promote the apoptosis of chicken kidneys, while the above changes were significantly alleviated in the ASB/Cd group. The results showed that ASB antagonizes the negative effects of Cd and against Cd-induced apoptosis in chicken kidneys via ERS signaling pathway.

Subambient daytime radiative cooling textile based on nanoprocessed silk.

Decreasing energy consumption is critical to sustainable development. Because temperature regulation for human comfort consumes vast amounts of energy, substantial research efforts are currently directed towards developing passive personal thermal management techniques that cool the human body without any energy consumption1-9. Although various cooling textile designs have been proposed previously, textile-based daytime radiative cooling to a temperature below ambient has not been realized6-13. Silk, a natural protein fabric produced by moth caterpillars, is famous for its shimmering appearance and its cooling and comforting sensation on skin14-17. It has been recently recognized that silk, with its optical properties derived from its hierarchical microstructure, may represent a promising starting point for exploring daytime radiative cooling18-21. However, the intrinsic absorption of protein in the ultraviolet region prevents natural silk from achieving net cooling under sunlight. Here we explore the nanoprocessing of silk through a molecular bonding design and scalable coupling reagent-assisted dip-coating method, and demonstrate that nanoprocessed silk can achieve subambient daytime radiative cooling. Under direct sunlight (peak solar irradiance >900 W m-2) we observed a temperature of ~3.5 °C below ambient (for an ambient temperature of ~35 °C) for stand-alone nanoprocessed silks. We also observed a temperature reduction of 8 °C for a simulated skin when coated with nanoprocessed silk, compared with natural silk. This subambient daytime radiative cooling of nanoprocessed silk was achieved without compromising its wearability and comfort. This strategy of tailoring natural fabrics through scalable nanoprocessing techniques opens up new pathways to realizing thermoregulatory materials and provides an innovative way to sustainable energy.

Discovery of Novel and Potent N-Methyl-d-aspartate Receptor Positive Allosteric Modulators with Antidepressant-like Activity in Rodent Models.

N-Methyl-d-aspartate receptors (NMDARs) are glutamate-gated Na+ and Ca2+-permeable ion channels involved in excitatory synaptic transmission and synaptic plasticity. NMDAR hypofunction has long been implicated in the pathophysiology including major depressive disorders (MDDs). Herein, we report a series of furan-2-carboxamide analogues as novel NMDAR-positive allosteric modulators (PAMs). Through structure-based virtual screen and electrophysiological tests, FS2921 was identified as a novel NMDAR PAM with potential antidepressant effects. Further structure-activity relationship studies led to the discovery of novel analogues with increased potentiation. Compound 32h caused a significant increase in NMDAR excitability in vitro and impressive activity in the forced swimming test. Moreover, compound 32h showed no significant inhibition of hERG or cell viability and possessed a favorable PK/PD profile. Our study presented a series of novel NMDAR PAMs and provided potential opportunities for discovering of new antidepressants.

Quassinoids: Phytochemistry and antitumor prospect.

Quassinoids, originating from the oxidative degradation of tetracyclic tirucallane triterpene, are a diverse class of secondary metabolites identifying from nature mostly in Simaroubaceae family. The crucial pharmacological activities and structural complexity of quassinoids have long fascinated scientists due to their medicinal uses, infamous toxicity, and unique biosynthesis. In the past few decades, 482 quassinoids, assigned to 6 skeletons, have been isolated and identified from plants. The names, classes, molecular formula, and plant sources of these secondary metabolites are collated here. This review will be a detailed update of the naturally occurring quassinoids reported from the plant kingdom, providing an in-depth discussion of their diversity, antitumor activities, structure-activity relationship.

Illicium verum and Eucommia ulmoides leaf extracts promote nutrient availability and antioxidant capacity in piglets by upregulating duodenal and jejunal Nrf2/TNF-α.

The effect of Illicium verum extracts (IVE) or Eucommia ulmoides leaf extracts (ELE) on nutrient availability, duodenal and jejunal antioxidant ability of Duroc ×Landrace × Yorkshire (DLY) and Chinese native Licha-black (LCB) piglets was investigated. Ninety-six piglets (48 DLY and 48 LCB respectively) without significant difference in body weight (11.22 ± 0.32 kg) were used in a 2 × 4 factorial design. Animals were randomly allocated to four treatments, and each had four replicates with three DLY and three LCB piglets. Treatments were basal diet (CON) and basal diet with 500 mg/kg IVE, 250 mg/kg ELE and 50 mg/kg chlortetracycline (CHL) respectively. Animals were placed individually for 7-days adaptation following 42-days test. Results showed the significant interaction (p < 0.05) between dietary treatments and pig species in activity of total superoxide dismutase (T-SOD) and glutathione peroxidase (GSH-Px), content of malondialdehyde (MDA), and α-tumour necrosis factor (TNF-α), nuclear factor erythroid 2-related factor 2 (Nrf2) and Nrf2/TNF-α at mRNA and protein level in duodenum and jejunum of DLY and LCB piglets. The IVE and ELE increased (p < 0.05) activity of GSH-Px and T-SOD, and the Nrf2/TNF-α at mRNA and protein level, however, the decreased (p < 0.05) MDA content, and TNF-α at mRNA and protein level in duodenum and jejunum were observed. The CHL decreased (p < 0.05) activity of GSH-Px and T-SOD, TNF-α and Nrf2 at mRNA and protein level in duodenum, but increased (p < 0.05) MDA content and Nrf2/TNF-α in jejunum. DLY piglets had higher (p < 0.05) nutrient digestibility (organic matter, crude protein and gross energy), availability (biological value and net protein utilization), MDA content, and TNF-α at mRNA and protein level in jejunum, and had lower (p < 0.05) activity of GSH-Px and T-SOD, and Nrf2 and Nrf2/TNF-α at mRNA and protein level in duodenum and jejunum than LCB. In conclusion, the 500 mg/kg IVE and 250 mg/kg ELE improved the nutrient availability, and the improvement of antioxidant capacity is realized by activating the Nrf2/TNF-α of duodenum and jejunum. The CHL had adverse effects on antioxidant ability of DLY and LCB piglets. The results showed that the nutrient digestion and absorption capacity of DLY were stronger than that of LCB piglets, but the antioxidant capacity was lower than that of LCB piglets. Therefore, the IVE and ELE are recommended as a new potential alternative to antibiotics in piglets' diets.

High-resolution X-ray structure of three microtubule-stabilizing agents in complex with tubulin provide a rationale for drug design.

Microtubule is a key component of cytoskeleton and has been considered as an important target for the treatment of cancer. In particular, the tubulin taxane-site inhibitors such as taxol analogs and epothilones have achieved great success in clinical trials. However, the structural basis of many taxane-site inhibitors is still lacking in exploring their mechanism of action. We here reported crystal complex structures for three taxane-site inhibitors, Ixabepilone, Epothilone B, and Epothilone D, which were determined to 2.4 Å, 2.4 Å, and 2.85 Å, respectively. The crystal structures revealed that these taxane-site inhibitors possess similar binding modes to that of Epothilone A at the taxane site, e.g. making critical hydrogen-bonding interactions with multiple residues on the M-loop, which facilitating the tubulin polymerization. Furthermore, we summarized the binding modes of almost all taxane-site inhibitors and identified novel taxane-site ligands with simpler chemical structures through virtual screening. On this basis, new derivatives with higher binding affinity to tubulin were designed and developed, which can form additional hydrogen bond interactions with tubulin. Overall, this work determined the mechanism of action of epothilones and provided a structural basis to design reasonably novel taxane-site inhibitors with simpler structure and improved pharmacokinetic properties.

A new dilignan from the twigs and leaves of Archidendron clypearia.

Previous work has shown that the lignans from the twigs and leaves of Archidendron clypearia (Jack) I.C.N. possess anti-ß-amyloid aggregation activity. Here we report a new dilignan, archidendronin A (1), along with one known sesquilignan (2). Their structures were determined by extensive spectroscopic methods, including UV, HRESIMS, 1 D and 2 D NMR data. The inhibitory activity on Aß1-42 aggregation was screened by ThT assay with curcumin as the positive control, and compounds 1 and 2 showed inhibition rate of 60.0% and 64.4% at 20 µM, respectively.[Formula: see text].

A polysaccharide extracted from alfalfa activates splenic B cells by TLR4 and acts primarily via the MAPK/p38 pathway.

Alfalfa polysaccharide (APS) has been proposed to exhibit growth-promoting and immune-enhancing bodily functions in vivo. However, little is known about its downstream immunomodulatory and intrinsic molecular mechanisms. Herein, mouse splenic lymphocytes were isolated to characterize the immunomodulatory effects and molecular mechanisms of APS in vitro. The results demonstrated that APS selectively improved the cell viability and IgM production of B cells, but no effects on T cell viability or secretion of IL-2, IL-4 and IFN-γ were observed in vitro. The receptor blocking assay showed that TLR4 was the primary receptor involved in APS-mediated B cell activation, which was confirmed by the results obtained using C57BL/10ScNJ (TLR4 gene-deficient) mice. Moreover, APS activated the TLR4-MyD88 signaling pathway at the translational level by significantly increasing the protein expression of TLR4 and MyD88. Downstream pathway blocking assay demonstrated that both the MAPK and NF-κB pathways were involved in APS-induced B cell activation. Additionally, APS significantly enhanced the phosphorylation of p38, ERK, and JNK and activated the nuclear translocation of the NF-κB p65 subunit. Therefore, we concluded that APS specifically activates the immune functions of splenic B cells by TLR4, acting through the MAPK and NF-κB signaling pathways, and potently activates the p38 pathway.

Lignans and neolignans with isovaleroyloxy moiety from Solanum lyratum Thunb.: Chiral resolution, configurational assignment and neuroprotective effects.

Eight pairs of enantiomeric lignans and neolignans including thirteen undescribed compounds, along with an undescribed meso compound, were isolated from the herbs of Solanum lyratum Thunb.(Solanaceae). Their structures and relative configurations were determined by extensive spectroscopic analyses of HRESIMS and nuclear magnetic resonance. The absolute configurations of the pure isomers were established based on the cooperative comparison between the experimental and calculated electronic circular dichroism (ECD) and optical rotation (OR). It is interesting that we obtained several naturally occurring stereoisomers with the identical gross structure possessing several stereogenic carbons from S. lyratum. Additionally, all isolates were assessed for neuroprotective effects toward human neuroblastoma SH-SY5Y cells injury induced by H2O2.

Effects of Jianpi Yiqi method for chronic obstructive pulmonary disease: A protocol for systematic review and meta-analysis.

BACKGROUND: Chronic obstructive pulmonary disease (COPD) is a chronic inflammatory lung disease with high morbidity and mortality worldwide. Jianpi Yiqi method (JYM) is a classical Chinese therapy in the treatment of COPD. However, there is no systematic review related to JYM for COPD. In this study, we aim to systematically examine the efficacy and safety for clinical use of JYM. METHODS AND ANALYSIS: We will conduct a comprehensive retrieval in the following electronic databases: PubMed, Embase, MEDLINE, Cochrane Library Central Register of Controlled Trials, China National Knowledge Infrastructure (CNKI) database, Wanfang Data Knowledge Service Platform, Chinese Scientific Journals Database (VIP), Chinese Biomedical Literature Service System (SinoMed), and other sources. Two trained reviewers will identify relevant studies, extract data information, and then assess the methodical quality by the Cochrane risk of bias assessment tool, independently. Then the meta-analyses will be conducted by using the RevMan 5.2. Based on the heterogeneity test, data integration is performed using a fixed effect model or a random effects model. A sensitivity analysis will be performed to evaluate the stability of the results. Then publication bias assessment will be conducted by funnel plot analysis and Egger test. Finally, the quality of evidence will be assessed by the GRADE system. RESULTS: The results of our research will be published in a peer-reviewed journal. CONCLUSION: The conclusion of our systematic review will provide evidence to judge whether JYM is an effective intervention for patients with COPD. OSF REGISTRATION NUMBER: 10.17605/OSF.IO/JKQYV.

Terpenylated coumarins from the root bark of Ailanthus altissima (Mill.) Swingle.

Seven undescribed terpenylated coumarins, named altissimacoumarin I-O, together with seven known compounds, altissimacoumarin C, altissimacoumarin E, altissimacoumarin G, altissimacoumarin H, puberulin, 7-(3-Methyl-2-butenyloxy)-6-methoxycoumarin and artelin were isolated from the root bark of Ailanthus altissima (Mill.) Swingle. Their structures were elucidated by comprehensive spectra data analysis, NMR calculation, DP4+ analysis and ACD/Structure Elucidator software simulation. The absolute configurations of altissimacoumarins K, L, M and N were determined by modified Mosher's method. All isolates were tested for their cytotoxic effect against two hepatoma carcinoma cell lines (HepG2, Hep3B). Altissimacoumarin C exhibited moderate cytotoxic effect against Hep3B cells, with IC50 of 45.21 µM.

Illicium verum extracts and probiotics with added glucose oxidase promote antioxidant capacity through upregulating hepatic and jejunal Nrf2/Keap1 of weaned piglets.

Accumulating evidences indicate that plant extracts and probiotics are effective antioxidant substitutes which play important roles in animal production. However, the comparative study of the mechanism underlying the antioxidant property of Illicium verum extracts (IVE) and probiotics with added glucose oxidase (PGO) on piglets remains to be explored. This study evaluated the difference and the interaction effect of IVE and PGO on serum, liver, and jejunum antioxidant capacity of weaned piglets. A total of 32 weaned piglets (Duroc × Landrace × Yorkshire) at the age of 28 d with an average body weight of 14.96 ± 0.32 kg were randomly divided into four treatments with eight replicates per treatment in a 2 × 2 factorial arrangement. Treatments included basal diet (IVE-PGO-), basal diet + 1,000 mg/kg PGO (IVE-PGO+), basal diet + 500 mg/kg IVE (IVE+PGO-), and basal diet + 500 mg/kg IVE + 1,000 mg/kg PGO (IVE+PGO+). All the piglets were housed individually for the 42-d trial period after 7-d adaptation. The piglets were euthanized at the end of the experiment and the liver and jejunum samples were taken and subjected to immunohistochemistry, Western blotting, as well as antioxidant and qRT-PCR analysis. Significant interactions were observed between IVE and PGO for total superoxide dismutase (T-SOD) and glutathione peroxidase (GSH-Px) in serum (42 d), liver, and jejunum; malondialdehyde (MDA) in serum (21 d); and mRNA and protein expression of kelch sample related protein-1 (Keap1) and nuclear factor erythroid-2 related factor (Nrf2)/Keap1 in the liver and jejunum (P < 0.05). Both IVE and PGO improved (P < 0.05) T-SOD and GSH-Px in the serum (42 d), liver, and jejunum, and the mRNA and protein expression of Nrf2 and Nrf2/Keap1 in the liver and jejunum, but decreased (P < 0.05) MDA in the serum (21 d) and the mRNA and protein expression of Keap1 in the liver and jejunum. Immunohistochemical results confirmed that IVE and PGO enhanced the positive reactions of Nrf2 but weakened Keap1 in both the liver and jejunum. In conclusion, the results confirmed that IVE (500 mg/kg) and PGO (1,000 mg/kg) can improve the antioxidant capacity of weaned piglets and that the interaction effect between IVE and PGO is significant. At the same time, the fact that IVE and PGO activate the Nrf2/Keap1 in the liver and jejunum signaling pathway suggests that they play an important role in the ameliorative antioxidant capacity of weaned piglets. Therefore, the combination of IVE and PGO could be recommended as a new potential alternative to antibiotics in piglets' diets.