In Vivo Anti-Hepatocellular Carcinoma Effects of the Chloroform Root Extract of Clausena excavata Burm.

Liver cancer is the most common cancer among males in Africa. The disease has a poor prognosis and its treatment is associated with toxicity and resistance. For this reason, numerous herbal combinations are being subjected to anticancer screening to circumvent the shortcomings of the conventional anticancer drugs. In the current study, the in vivo anti-cancer effects of the chloroform root extract of the herb, Clausena excavata Burm were investigated. Liver cancer was induced in mice by a single intraperitoneal injection of diethylnitrosamine (DEN) followed by oral administration of the promoter of carcinogenesis, 2-aminoacetyl fluorine that was mixed with the mice feed. The cytotoxicity of the root extract of C. excavata on liver cancer cells was investigated using liver enzyme, histology, DNA fragmentation and caspases assays. Real time qPCR was conducted to evaluate the effect of the extract on apoptotic genes. The findings revealed that the extract of C. excavata significantly decreased the progression of hepatocarcinogenesis and the toxicity-induced production of the liver enzymes, alanine and aspartate aminotransferases. The histological analyses of the liver tissues revealed evidence of apoptotic cell death. The extract also provoked significant (p < .05) expressions of caspase 9 protein and gene as well as other apoptotic genes (P53, P27, Apaf-1, cytochrome C, bax and bid). Therefore, we postulate that the chloroform root extract of C. excavata induces apoptosis of liver cancer in mice.

Chemical profile of the roots of Clausena lansium and their inhibitory effects of the over-activation in BV-2 microglial cells.

From the 95% ethanol aqueous extract of the roots of Clausena lansium, six previously undescribed alkaloids (1, 2a, 2b, 15, 24a, 24b), a pair of prenylated phenylpropenols (26a, 26b), two coumarins (27, 28), and two undescribed sesquiterpenes (37, 38) were isolated and identified using spectroscopic and electron circular dichroism data, together with thirty-two known compounds. The absolute configurations of three alkaloids (3a, 3b, 4a) were determined for the first time. In vitro assay showed that alkaloids 7, 10, 12, 19, and furanocoumarins 34, 35 displayed inhibitory effects on the production of nitric oxide in lipopolysaccharide (LPS)-induced BV-2 microglial cells, which were stronger than that of the minocycline (positive control). RT-PCR results indicated that indizoline (7) could inhibit the expression of pro-inflammatory factors (IL-1ß, TNF-α, and IL-6) in LPS-treated BV-2 cells.

Structural characterization of a water-soluble acidic polysaccharide CSP-IV with potential anticoagulant activity from fruit pulp of Clausena lansium (Lour.) Skeels Guifei.

Four main water-soluble wampee fruit pulp polysaccharides, named CSP-I, CSP-II, CSP-III and CSP-IV, were isolated from Clausena lansium (Lour.) Skeels Guifei, therein CSP-IV content was higher than the others. All components possess certain anticoagulant activity demonstrated by prolonged activated partial thromboplastin time, especially CSP-IV, which suggests that CSP-IV plays anticoagulant effect through disturbing intrinsic coagulation pathway. The wampee polysaccharide CSP-IV with Mw of 510.1 kDa was mainly composed of Gal, Ara and GalA. Backbone of CSP-IV contains Gal, Ara and GalA, two kinds of side chains contain one monosaccharide Gal or Ara, both branch on Gal residue of backbone. CSP-IV has no the conformation of triple helix demonstrated by Congo red test. These results showed that CSP-IV is an acidic polysaccharide with potential anticoagulant activity via targeting intrinsic coagulation pathway.

RNA-Seq-Based Transcriptomics and GC-MS Quantitative Analysis Reveal Antifungal Mechanisms of Essential Oil of Clausena lansium (Lour.) Skeels Seeds against Candida albicans.

Infections caused by Candida albicans (C. albicans) and increasing resistance to commonly used drugs lead to a variety of mucosal diseases and systemic infectious diseases. We previously confirmed that the essential oil of Clausena lansium (Lour.) Skeels seeds (CSEO) had antifungal activity against C. albicans, but the detailed mechanism between the chemical components and antifungal activity is unclear. In this study, a quantitative analysis of five volatile components of CSEO, including sabinene, α-phellandrene, ß-phellandrene, 4-terpineol, and ß-caryophyllene, was carried out using the gas chromatography-mass spectrometry (GC-MS) method. Both the broth dilution and kinetic growth methods proved that the antifungal activity of CSEO against fluconazole-resistant C. albicans was better than that of its main components (sabinene and 4-terpineol). To further investigate the inhibitory mechanism, the transcriptional responses of C. albicans to CSEO, sabinene, and 4-terpineol treatment were determined based on RNA-seq. The Venn diagram and clustering analysis pattern of differential expression genes showed the mechanism of CSEO and 4-terpineol's anti-C. albicans activity might be similar from the perspective of the genes. Functional enrichment analysis suggested that CSEO regulated adherence-, hyphae-, and biofilm-formation-related genes, which may be CSEO's active mechanism of inhibiting the growth of fluconazole-resistant C. albicans. Overall, we preliminarily revealed the molecular mechanism between the chemical components and the antifungal activity of CSEO against C. albicans. This study provides new insights to overcome the azole resistance of C. albicans and promote the development and application of C. lansium (Lour.) Skeels seeds.

Chitosan-based edible film incorporated with wampee (Clausena lansium) seed essential oil: Preparation, characterization and biological activities.

Chitosan (Ch)-based edible composite films were prepared by incorporating blending wampee seed essential oil (WSEO) into a Ch matrix, using the incorporation ratio as a variable. The physical, mechanical properties, structure morphology and rheological properties were determined using tensile strength (TS), elongation at break (EB), water vapor permeability (WVP) tests together with Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM) observations and apparent viscosity and shear rate. In addition, the antimicrobial, antioxidant activities were investigated by the DPPH & ABTS radicals scavenging and inhibition zone assays, respectively. Compared with Ch, the incorporation of WSEO significantly decreased (P < 0.05) the TS, EB, and WVP values, especially when the WSEO ratio reached 1.0 % or higher. Meanwhile, the films exhibited greatly improved visible light barrier performance after WSEO incorporation. Both FTIR spectroscopy and SEM observations reflected the crosslinking between WSEO and Ch. Meanwhile, the composite films demonstrated smaller particle size and weaker rheological viscosities, which enhanced the antimicrobial and antioxidant capabilities when compared with those of Ch. Therefore, this study suggested that WSEO incorporated with Ch is an effective ingredient for the preparation of edible films with enhanced physicochemical and biological properties.

The primary neurotoxic factor, Lansamide I, from Clausena lansium fruits and metabolic dysfunction invoked.

Wampee (Clausena lansium) is a common fruit in South Asia. The pulp is a tasty food, and the seed is a typical traditional herb in China. However, we identified a primary toxic compound, Lansamide I, by NMR and X-ray diffraction of single-crystal. The compound occurred at 4.17 ± 0.16 mg/kg of dried seed and 0.08 ± 0.01 g/kg of fresh fruit. In our phenotype-based toxicity investigation, the compound caused decreased hatchability of zebrafish eggs, increased malformations such as enlarged yolk sacs and pericardial edema, and delayed body length development. Moreover, the compound also caused nerve cell damage and decreased locomotor activity. The compound caused an increase in peroxidation levels in vivo, with increases in both malondialdehyde and superoxide dismutase levels, but did not interfere with acetylcholinesterase levels. Metabolomic studies found that the compound caused significant up-regulation of 16 metabolites, mainly amino acids and peptides, which were involved in the nucleotide metabolism pathway and the betaine biosynthesis module. The qRT-PCR revealed that the substance interfered with the mRNA expression of tat and dctpp. These discoveries offer fresh perspectives on the toxicity mechanisms and metabolic response to the primary harmful molecules in wampee, which could inform the rational usage of wampee resources.

Phenolic profile and α-glucosidase inhibitory potential of wampee (Clausena lansium (Lour.) Skeels) peel and pulp: In vitro digestion/in silico evaluations.

To investigate the changes in phenolics, flavonoids, and their bio-activities of wampee (Clausena lansium (Lour.) Skeels) during digestion, the peel and pulp were subjected to simulated in vitro digestion, encompassing oral, gastric, small intestine, and large intestine digestion stages. The peel exhibited a total release of 91.93 mg GAE/g DW of phenolics and 61.86 mg RE/g DW of flavonoids, whereas the pulp displayed a release of 27.83 mg GAE/g DW of phenolics and 8.94 mg RE/g DW of flavonoids. Notably, the phenolics and flavonoids were mostly released during the oral digestion stage for peel, while they were mostly released during the small intestine digestion stage for pulp. The results of the targeted flavonoids analysis indicated that rutin and l-epicatechin were the two most widely released compounds in each digestion step. Moreover, myricetin has been identified as the best inhibitor against α-glucosidase, probably because it formed the most H-bonds, 8, with 6 catalytic residues, which was the highest number. Furthermore, the soluble substances released from the peel exhibited significantly higher antioxidant activities and inhibitory activity against α-glucosidase (p < 0.05) compared to those from the pulp. Positive correlations were observed between the total phenolic content or total flavonoid content and the antioxidant activities (r > 0.73 (peel), > 0.61 (pulp)), as well as α-glucosidase inhibitory activity (r <  - 0.48 (peel), < -0.64 (pulp)) of peel and pulp. In conclusion, these findings provide valuable insights into the digestive characteristics and health benefits of both wampee peel and pulp.

Anti-malarial activity of the alkaloid, heptaphylline, and the furanocoumarin, imperatorin, from Clausena anisata against human Plasmodium falciparum malaria parasites: ex vivo trophozoitocidal, schizonticidal and gametocytocidal approach.

BACKGROUND: The erythrocytic stage of the life cycle of the malaria parasite, Plasmodium falciparum, consists of trophozoite, schizont and gametocyte stages in humans. Various anti-malarial agents target different stages of the parasite to produce treatment outcomes. This study reports on the stage-specific anti-malarial activity of heptaphylline and imperatorin against human P. falciparum in addition to their cytotoxicity and selectivity indices (SI). METHODS: The compounds were isolated from Clausena anisata using column chromatography and their structures elucidated using NMR spectroscopy. The anti-malarial activity was determined by measuring the trophozoitocidal, schizonticidal and gametocytocidal activities of the compounds using the SYBR green assay. Cytotoxicity was evaluated using the tetrazolium-based colorimetric assay. RESULTS: Heptaphylline and imperatorin produced trophozoitocidal, schizonticidal and gametocytocidal activities with IC50s of 1.57 (0.2317)-26.92 (0.3144) µM with those of artesunate (the standard drug) being 0.00024 (0.0036)-0.0070 (0.0013) µM. In the cytotoxicity assay, the compounds produced CC50S greater than 350 µM and SI of 13.76-235.90. Also, the trophozoitocidal and schizonticidal activities of the compounds were more pronounced than their gametocytocidal activity. Imperatorin was 42.04% more trophozoitocidal than hepthaphyline. However, hepthaphyline has more schizonticidal and gametocytocidal properties than imperatorin. CONCLUSION: Heptaphylline and imperatorin are promising anti-malarial agents, since they possess potent anti-malarial activity with weak cytotoxicity on RBCs. However, imperatorin is a better anti-malarial prophylactic agent whereas heptaphylline is a better malaria treatment agent.

New Phenolic Lipids from the Leaves of Clausena harmandiana Inhibit SARS-CoV-2 Entry into Host Cells.

Induced by the spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the COVID-19 pandemic underlined the clear need for antivirals against coronaviruses. In an effort to identify new inhibitors of SARS-CoV-2, a screening of 824 extracts prepared from various parts of 400 plant species belonging to the Rutaceae and Annonaceae families was conducted using a cell-based HCoV-229E inhibition assay. Due to its significant activity, the ethyl acetate extract of the leaves of Clausena harmandiana was selected for further chemical and biological investigations. Mass spectrometry-guided fractionation afforded three undescribed phenolic lipids (1-3), whose structures were determined via spectroscopic analysis. The absolute configurations of 1 and 2 were determined by analyzing Mosher ester derivatives. The antiviral activity against SARS-CoV-2 was subsequently shown, with IC50 values of 0.20 and 0.05 µM for 2 and 3, respectively. The mechanism of action was further assessed, showing that both 2 and 3 are inhibitors of coronavirus entry by acting directly on the viral particle. Phenolic lipids from Clausena harmandiana might be a source of new antiviral agents against human coronaviruses.

Bioassay-Guided Isolation and Identification of Antiplasmodial Compounds from the Stem Bark of Clausena excavata.

Clausena excavata is a medicinal plant widely distributed in Southeast Asia. It is used for a variety of indications, including to treat malaria. In our present study, a phytochemical study of the methanol extract from the stem bark of C. excavata led to the isolation of five pyranocoumarins, nordentatin (1: ), dentatin (2: ), kinocoumarin (3: ), clausarin (4: ), and clausenidin (5: ), and a coumarin, 8-hydroxy-3″,4″-dihydrocapnolactone-2',3'-diol (6: ). The isolation of compound 6: from C. excavata and the antiplasmodial activities against a multidrug-resistant K1 strain of Plasmodium falciparum of 1, 3: , and 5: were reported for the first time. Compounds 3: and 4: exhibited potent antiplasmodial activities with EC50 values of 1.10 and 0.58 µM, respectively, while 1: and 5: had EC50 values of 5.62 and 7.15 µM, respectively. A prenyl group attached to the C-3 or C-12 position on the pyranocoumarin ring probably plays an important role on the activity. A hydroxyl group at the C-10 position is also likely to enhance the activity.

Ethyl acetate extract of Clausena execavata promotes growth inhibition of Burkitt's lymphoma cell line via apoptotic activities / El extracto de acetato de etilo de Clausena excavata promueve la inhibición del crecimiento de la línea celular del linfoma de Burkitt a través de actividades apoptóticas

Clausena excavata is a famous folklore medicinal plant in Asian region that is being used for the treatment of different disorders. This study investigated the cytotoxic effects of leaf extracts via MTT assay, as well as the in vitro apoptotic activities of the ethyl acetate C. excavata leaf extract (EACE) on human Burkitt's lymphoma, Raji, cell line using annexin-V-FITC/propidium iodide flow cytometric assays. Pro-apoptotic (BAX) and anti-apoptotic (BCL-2, c-MYC) gene expressions were determined via real-time quantitative PCR. Phytochemical screening was done by Gas chromatography-mass spectrometry (GC-MS). EACE has the lowest IC50 (19.3 ± 0.35 µg/mL) among extracts. EACE-treated Raji cells after 48 h underwent apoptosis as evident by loss of cell viability and increase in the percentage of early and late apoptotic cells. The results also showed EACE mediated decreased in the BCL-2 and c-MYC gene expressions and increased in the BAX gene. C. excavata is a potential treatment for Burkitt lymphoma through activation of apoptosis.

Clausena excavata es una planta medicinal tradicional famosa en la región asiática que se utiliza para el tratamiento de diferentes trastornos. Este estudio investigó los efectos citotóxicos de los extractos de hojas a través del ensayo MTT, así como las actividades apoptóticas in vitro del extracto de hoja de acetato de etilo de C. excavata (EACE) en la línea celular de linfoma de Burkitt humano, Raji, usando citometría de flujo de yoduro de anexina-V-FITC/propidio. Las expresiones génicas proapoptóticas (BAX) y antiapoptóticas (BCL-2, c-MYC) se determinaron mediante PCR cuantitativa en tiempo real. El cribado fitoquímico se realizó mediante cromatografía de gases-espectrometría de masas (GC-MS). EACE tiene el IC50más bajo (19,3 ± 0,35 µg/mL) entre los extractos. Las células Raji tratadas con EACE después de 48 h sufrieron apoptosis como es evidente por la pérdida de viabilidad celular y el aumento en el porcentaje de células apoptóticas tempranas y tardías. Los resultados también mostraron una disminución mediada por EACE en las expresiones de los genes BCL-2 y c-MYC y un aumento en el gen BAX. C. excavata es un tratamiento potencial para el linfoma de Burkitt a través de la activación de la apoptosis.

CZK, a novel alkaloid derivative from Clausena lansium, alleviates ischemic stroke injury through Nrf2-mediated antioxidant effects.

Anti-oxidant stress is a potential strategy for the treatment of ischemic stroke. Here, we found a novel free radical scavenger termed as CZK, which is derived from alkaloids contained in Clausena lansium. In this study, we first compared cytotoxicity and biological activity between CZK and its parent's compound Claulansine F. It was found that CZK had lower cytotoxicity and improved anti-oxygen-glucose deprivation/reoxygenation (OGD/R) injury than its parent's compound. Free radical scavenging test showed that CZK had a strong inhibitory effect on hydroxyl free radicals with the IC50 of 77.08 nM. Intravenous injection of CZK (50 mg/kg) significantly alleviated ischemia-reperfusion injury, manifested by reduced neuronal damage and decreased oxidative stress. Consistent with the findings, the activities of superoxide dismutase (SOD) and reduced glutathione (GSH) were increased. Molecular docking predicted that CZK might be combined with nuclear factor erythroid 2-related factor 2 (Nrf2) complex. Our results also confirmed that CZK upregulated the contents of Nrf2 and its target gene products Heme Oxygenase-1 (HO-1), and NAD(P)H: Quinone Oxidoreductase 1 (NQO1). In conclusion, CZK had a potential therapeutic effect for ischemic stroke by activating Nrf2-mediated antioxidant system.

Antifungal alkaloids from the branch-leaves of Clausena lansium Lour. Skeels (Rutaceae).

BACKGROUND: The rational utilization of botanical secondary metabolites is one of the strategies to reduce the application of chemical fungicides. The extensive biological activities of Clausena lansium indicate that it has the potential to develop botanical fungicides. RESULTS: A systematic investigation on the antifungal alkaloids from C. lansium branch-leaves following bioassay-guided isolation was implemented. Sixteen alkaloids, including two new and nine known carbazole alkaloids, one known quinoline alkaloid and four known amides, were isolated. Compounds 4, 7, 12 and 14 showed strong antifungal activity on Phytophthora capsiciwith EC50 values ranging from 50.67 to 70.82 µg mL-1 . Compounds 1, 3, 8, 10, 11, 12 and 16 displayed different degrees of antifungal activity against Botryosphaeria dothidea with EC50 values ranging from 54.18 to 129.83 µg mL-1 . It was reported for the first time that these alkaloids had antifungal effects on P. capsici or B. dothidea, and their structure-activity relationships were further discussed systematically. Additionally, among all alkaloids, dictamine (12) had the strongest antifungal activities against P. capsici (EC50  = 50.67 µg mL-1 ) and B. dothidea (EC50  = 54.18 µg mL-1 ), and its physiological effects on P. capsici and B. dothidea also were further evaluated. CONCLUSION: Capsicum lansium is a potential source of antifungal alkaloids, and C. lansium alkaloids had the potential as lead compounds of botanical fungicides in the development of new fungicides with novel action mechanism. © 2023 Society of Chemical Industry.

Complete Genome Sequence Resource of Pantoea anthophila CL1 Causing Soft Rot Disease in Clausena lansium (Wampee) in China.

Pantoea anthophila CL1 is a causal agent of soft rot disease in Clausena lansium (wampee) in China and has inhibitory activity against the bacterial wilt pathogen Ralstonia solanacearum. Here we report the genome sequencing and analysis of P. anthophila CL1, representing the first complete genome resource of the species. The CL1 genome consists of four circular replicons (one chromosome and three plasmids), with a total size of 4,594,065 bp, and contains 4,109 protein-coding genes and 106 RNA genes. Our bioinformatic analysis of CL1 predicted 228 virulence factors, two Type VI Secretion Systems, and six secondary metabolite biosynthesis gene clusters producing saccharides, siderophores, and terpene. The complete genome sequence of P. anthophila CL1 provides a solid foundation for further investigation of its pathogenesis and antimicrobial activity and also represents a valuable resource for the comparative genomics of Pantoea.

Anethole rich Clausena heptaphylla (Roxb.) Wight & Arn., essential oil pharmacology and genotoxic efficiencies.

Anethole, a widely used industrial flavoring agent is majorly sourced from anise and star anise. The present study is aimed to the in-depth pharmacological analysis i.e. anti-diabetic, skin whitening, neurodegenerative disorder inhibitory activities of anethole-rich Clausena heptaphylla leaf essential oil (ARCHEO) (88.59%) as revealed by the Gas Chromatography/Mass Spectrometry (GC/MS) analysis and further confirmed by proton nuclear magnetic resonance 1H-NMR as well as to compare with standard compound anethole. ARCHEO (ABTS EC50 6.97 ± 0.004 µg/mL; Protease assay 4.51 ± 0.004 µg/mL) outperformed the standard compound anethole (ABTS EC50 9.48 ± 0.048 µg/mL; Protease assay EC50 22.64 ± 0.016 µg/mL) in antioxidant and anti-inflammatory experiments. ARCHEO was also shown to be more effective than the reference compound anethole in terms of anti-diabetic activity (EC50 22.35 ± 0.121 µg/mL), tyrosinase inhibitory activity (EC50 16.45 ± 0.012 µg/mL), and anti-cholinesterase activity (EC50 22.32 ± 0.016 µg/mL). However, ARCHEO exhibited lower antimicrobial activity towards all the tested microbes compared to standard compound anethole and as for the MIC, ARCHEO was effective only towards Salmonella typhimurium (60 µg/mL), Streptococcus mutans (20 µg/mL), and Aspergillus fumigatus (75 µg/mL). ARCHEO (11.11%) and anethole (12.33%) showed no genotoxic effect based on Allium cepa assay mitotic index value. Thus, ARCHEO could be a commercially viable and widely available cheaper source of anethole, which has buoyant demand in the field of food flavoring, fragrance, and pharmaceutical industries.

Physicochemical, structural, and functional properties of wampee (Clausena lansium (Lour.) Skeels) fruit peel pectin extracted with different organic acids.

Effects of different extraction acids on physicochemical, structural, and functional properties of wampee fruit peel pectin (WFPP) were comparatively investigated. The hydrochloric acid extracted WFPP (HEP) exhibited the highest degrees of methylation (67.79%) and acetylation (86.29%) coupling with abundant monosaccharides and rhamnogalacturonan branches, but lowest molecular weight (5.58 × 105 Da). The results of SEM, X-ray diffraction, and Fourier transform infrared spectroscopy analyses showed that acid types had little effect on the surface morphology of WFPP. However, compared to commercial citrus pectin (CCP), several specific absorbance peaks (1539, 1019, 920 cm-1) were found in WFPPs, which corresponds to aromatic skeletal stretching, pyranose, and d-glucopyranosyl, respectively. Moreover, the rheological behavior revealed that WFPP solution was pseudoplastic fluid and affected by acid types. And the WFPPs exhibited higher emulsifying activity and emulsion stability than CCP. All these WFPPs presented well antioxidant activity and promoting probiotics ability, especially for HEP.

Clausena Harmandiana root extract attenuated cognitive impairments via reducing amyloid accumulation and neuroinflammation in Aß1-42-induced rats.

BACKGROUND: Alzheimer's disease (AD) pathogenesis is associated with amyloid-ß (Aß)-induced neuroinflammation. In AD, the activation of microglia caused by Aß accumulation is followed by the synthesis and release of pro-inflammatory cytokines, including interleukin-1ß (IL-1ß) and tumor necrosis factor-α (TNFα), and ultimately leads to cognitive impairments. Clausena harmandiana (CH) is a medicinal plant in the Rutaceae family and has been used in folk medicine to relieve illnesses such as stomachache and headache, and as a health tonic. Interestingly, CH root extract (CHRE) has several anti-inflammatory and other pharmacological activities, but there are no studies in AD-like animal models. OBJECTIVES: This study aims to evaluate the effects of CHRE on cognitive impairments, increased Aß1-42 protein levels, and neuroinflammation in Aß1-42-induced rats. METHODS: Forty-eight adult male Sprague-Dawley rats (250-300 g) were randomly divided into 6 groups (n = 8) of the sham control, V + Aß, CB + Aß CHRE125 + Aß, CHRE250 + Aß, and CHRE500 + Aß. Sodium carboxymethylcellulose, Celebrex (10 mg/kg BW) and CHRE (125, 250, and 500 mg/kg BW) were given orally or without any treatment for 35 days. On day 21, aggregated Aß1-42 at a concentration of 1 µg/µl were injected into both lateral ventricles (1 µl/side) of all treated rats, while sterilized normal saline were injected to untreated rats. Ten days later, the novel object recognition test was performed to assess their recognition memory. At the end of the test period, an overdose of thiopental sodium (120 mg/kg BW) and transcardial perfusion with 0.9% normal saline solution were used to euthanize all rats. Then Aß1-42 protein levels and the expression of inflammatory markers (CD11b-positive microglia, IL-1ß, and TNFα) were investigated in the cerebral cortex and hippocampus. RESULTS: Pretreatment with CHRE at all doses could attenuate short- and long-term impairments in recognition memory. Additionally, CHRE also inhibited the increase of Aß1-42 protein levels and the expression of inflammatory markers in both brain regions as well as receiving Celebrex. CONCLUSIONS: This suggests that preventive treatment of CHRE might be a potential therapy against cognitive impairments via reducing Aß1-42 protein levels and neuroinflammation caused by Aß1-42.

Anti-Neuroinflammatory Components from Clausena lenis Drake.

Clausena lenis Drake (C. lenis) is a folk medicinal herb to treat influenza, colds, bronchitis, and malaria. The 95% and 50% ethanol extract of C. lenis showed significant nitric oxide (NO) inhibition activity in BV-2 microglial cells stimulated by lipopolysaccharide (LPS). Bio-guided isolation of the active extract afforded five new compounds, including a chlorine-containing furoquinoline racemate, (±)-claulenine A (1), an amide alkaloid, claulenine B (2), a prenylated coumarin, claulenin A (3), a furocoumarin glucoside, clauleside A (4), and a multi-prenylated p-hydroxybenzaldehyde, claulenin B (5), along with 33 known ones. Their structures were determined via spectroscopic methods, and the absolute configurations of new compounds were assigned via the electronic circular dichroism (ECD) calculations and single-crystal X-ray diffraction analysis. Compounds 2, 23, 27, 28, 33, and 34 showed potent anti-neuroinflammatory effects on LPS-induced NO production in BV-2 microglial cells, with IC50 values in the range of 17.6-40.9 µM. The possible mechanism was deduced to interact with iNOS through molecular docking.

Anti-Diabetes, Anti-Gout, and Anti-Leukemia Properties of Essential Oils from Natural Spices Clausena indica, Zanthoxylum rhetsa, and Michelia tonkinensis.

Essential oils (EOs) of Clausena indica fruits, Zanthoxylum rhetsa fruits, and Michelia tonkinensis seeds were analyzed for their phytochemical profiles and biological activities, including anti-diabetes, anti-gout, and anti-leukemia properties. Sixty-six volatile compounds were identified by gas chromatography-mass spectrometry (GC-MS), in which, myristicin (68.3%), limonene (44.2%), and linalool (49.3%) were the most prominent components of EOs extracted from C. indica, Z. rhetsa, and M. tonkinensis, respectively. In addition, only EOs from C. indica inhibited the activities of all tested enzymes comprising α-amylase (IC50 = 7.73 mg/mL), α-glucosidase (IC50 = 0.84 mg/mL), and xanthine oxidase (IC50 = 0.88 mg/mL), which are related to type 2 diabetes and gout. Remarkably, all EOs from C. indica, Z. rhetsa (IC50 = 0.73 mg/mL), and M. tonkinensis (IC50 = 1.46 mg/mL) showed a stronger anti-α-glucosidase ability than acarbose (IC50 = 2.69 mg/mL), a known anti-diabetic agent. Moreover, the growth of leukemia cell Meg-01 was significantly suppressed by all EOs, of which, the IC50 values were recorded as 0.32, 0.64, and 0.31 mg/mL for EOs from C. indica, Z. rhetsa, and M. tonkinensis, respectively. As it stands, this is the first report about the inhibitory effects of EOs from C. indica and Z. rhetsa fruits, and M. tonkinensis seeds on the human leukemia cell line Meg-01 and key enzymes linked to diabetes and gout. In conclusion, the present study suggests that EOs from these natural spices may be promising candidates for pharmaceutical industries to develop nature-based drugs to treat diabetes mellitus or gout, as well as malignant hematological diseases such as leukemia.

Characterization and prebiotic properties of pectin polysaccharide from Clausena lansium (Lour.) Skeels fruit.

Pectins have proven to be advantageous for human health as they regulate beneficial microbial communities and enhance immunity. The fruit of Clausena lansium (Lour.) Skeels (Wampee), also referred to as "treasure in fruit", is rich in pectin polysaccharides. In this study, a homogalacturonan-type pectin (CCP2) with a molecular weight of 8.9 × 104 Da and degree of esterification of 42.86% was isolated from Wampee fruit. The gut microbiota regulation and phagocytosis-enhancing properties of CCP2 were examined in vivo and in vitro, respectively. Oral administration of CCP2 dramatically decreased the abundance of Bacteroidetes and increased the abundance of Firmicutes in intestinal bacteria in mice. The content of short-chain fatty acids in the feces also significantly improved. Moreover, CCP2 exhibited excellent phagocytosis-enhancing activities on RAW 264.7 macrophages. These results suggested that CCP2 could be a potential gut microbiota regulator and phagocytosis-enhancer, which could be used in food products to promote health through beneficial manipulation of gut microbiota.