BACKGROUND AND OBJECTIVE

Diabetes, a prevalent metabolic disorder characterized by hyperglycemia primarily due to insulin action and secretion, poses significant health challenges, particularly in low to medium-income countries such as the Philippines. Quassia amara, a shrub indigenous to South America and present in the Philippines, holds a rich history of utilization in alternative and complementary therapies. While previous studies have demonstrated the hypoglycemic effects of Quassia amara stem wood, investigations into the potential impact of its leaves on blood glucose levels remain scarce. Thus, this study aimed to assess the blood glucose-lowering effects of the aqueous leaf extract of Quassia amara (ALQa) on ICR strain mice.

Diabetes was induced in thirty male ICR mice via intraperitoneal administration of alloxan monohydrate (200 mg/kg) dissolved in 0.9% Normal Saline. The mice were divided into five groups (n=6), Group I: negative control (distilled water), Group II: reference standard glibenclamide (4 mg/kg): Groups III-V: three doses of ALQa (125, 250, and 500 mg/kg) via oral gavage. A glucometer was used to monitor the fasting blood glucose levels at 0, 1-, 2-, 6-, and 24-hour postadministration.

Administration of alloxan monohydrate increased the FBS in the treated group to diabetic levels of >200 mg/dL. The treatment of diabetic mice with ALQa extract significantly reduced fasting blood sugar (FBS) levels in a dose-dependent manner with the highest dose of ALQa (500 mg/kg) having glucoselowering effects comparable to glibenclamide beginning with the 2-hour mark until 24-hour post-intervention. The mean FBS at 0-hour (baseline) and 1-hour postintervention were similar for all the groups. However, there was an increase in the mean FBS of the negative control group treated with distilled water in the first hour while there was already a decrease in the FBS of those allocated to glibenclamide and the three doses of ALQa. At both the second and 6-hour mark post-intervention, the mean FBS of the mice treated with ALQa 250 mg/ kg and 500 mg/kg was comparable to glibenclamide. Finally, at the 24th hour post-intervention, only the mice allocated to 500 mg/kg of ALQa had comparable FBS to glibenclamide. The degree of reduction [mean percent reduction] of the FBS from baseline to the 24th hour was 78% for glibenclamide and 69% for ALQa 500 mg/kg (p =0.816).

The aqueous extract of Quassia amara leaf at 250 and 500 mg/kg produced a dose-dependent significant blood glucose-lowering effect in the alloxan-induced diabetic mice model. The 500 mg dose demonstrated a statistically comparable reduction in FBS to glibenclamide from the 2-hour time point. These f indings suggest the potential of ALQa as an antidiabetic agent. Thus, warranting further investigation into its therapeutic mechanisms and clinical applications.

Background and Objective: Diabetes, a prevalent metabolic disorder characterized by hyperglycemia primarily due to insulin action and secretion, poses significant health challenges, particularly in low to medium-income countries such as the Philippines. Quassia amara, a shrub indigenous to South America and present in the Philippines, holds a rich history of utilization in alternative and complementary therapies. While previous studies have demonstrated the hypoglycemic effects of Quassia amara stem wood, investigations into the potential impact of its leaves on blood glucose levels remain scarce. Thus, this study aimed to assess the blood glucose-lowering effects of the aqueous leaf extract of Quassia amara (ALQa) on ICR strain mice. Methods: Diabetes was induced in thirty male ICR mice via intraperitoneal administration of alloxan monohydrate (200 mg/kg) dissolved in 0.9% Normal Saline. The mice were divided into five groups (n=6), Group I: negative control (distilled water), Group II: reference standard glibenclamide (4 mg/kg): Groups III-V: three doses of ALQa (125, 250, and 500 mg/kg) via oral gavage. A glucometer was used to monitor the fasting blood glucose levels at 0, 1-, 2-, 6-, and 24-hour postadministration. Results: Administration of alloxan monohydrate increased the FBS in the treated group to diabetic levels of >200 mg/dL. The treatment of diabetic mice with ALQa extract significantly reduced fasting blood sugar (FBS) levels in a dose-dependent manner with the highest dose of ALQa (500 mg/kg) having glucoselowering effects comparable to glibenclamide beginning with the 2-hour mark until 24-hour post-intervention. The mean FBS at 0-hour (baseline) and 1-hour postintervention were similar for all the groups. However, there was an increase in the mean FBS of the negative control group treated with distilled water in the first hour while there was already a decrease in the FBS of those allocated to glibenclamide and the three doses of ALQa. At both the second and 6-hour mark post-intervention, the mean FBS of the mice treated with ALQa 250 mg/ kg and 500 mg/kg was comparable to glibenclamide. Finally, at the 24th hour post-intervention, only the mice allocated to 500 mg/kg of ALQa had comparable FBS to glibenclamide. The degree of reduction [mean percent reduction] of the FBS from baseline to the 24th hour was 78% for glibenclamide and 69% for ALQa 500 mg/kg (p =0.816). Conclusions The aqueous extract of Quassia amara leaf at 250 and 500 mg/kg produced a dose-dependent significant blood glucose-lowering effect in the alloxan-induced diabetic mice model. The 500 mg dose demonstrated a statistically comparable reduction in FBS to glibenclamide from the 2-hour time point. These f indings suggest the potential of ALQa as an antidiabetic agent. Thus, warranting further investigation into its therapeutic mechanisms and clinical applications.
Quassia amara ; Quassia

OBJECTIVE: To explore the mechanisms that mediate the anti-inflammatory activity of Eurycoma longifolia. METHODS: Kunming mouse models of xylene-induced ear swelling and lipopolysaccharide (LPS)-induced acute pneumonia were used to compare the anti- inflammatory activities of aqueous and ethanol extracts of Eurycoma longifolia. UPLC-Q-TOF-MS/MS was used to identify the chemical composition in the ethanol extract of Eurycoma longifolia, based on which the potential antiinflammatory targets of Eurycoma longifolia were screened using the databases including SwissADME, SwissTargetPrediction, and Genecards. The String database was used to generate the protein-protein interaction (PPI) network, and Cytoscape was used for network topology analysis and screening the core targets. The enrichment of the core targets was analyzed using Metascape database, the core components and targets were docked with Autodock software, and the docking results were visualized using Pymol software. In a RAW264.7 cell model of LPS-induced inflammation, the Griess reagent was used to measure NO level, and Western blotting was performed to detect the expression levels of MAPK1, JAK2, and STAT3 proteins to verify the anti- inflammatory mechanism of Eurycoma longifolia. RESULTS: The ethanol extract (75%) of Eurycoma longifolia (ELE) was the active site, which contained a total of 37 chemical components. These chemical compounds and diseases had 541 targets, involving the JAK/STAT3, cAMP and other signaling pathways. Twelve indicator components were identified, which all showed good results of molecular docking with two core targets involved in the signaling pathways. In the cell validation experiment, treatment of the cells with low-, medium-, and high-dose ELE significantly reduced NO release in the cells, and ELE at the medium dose significantly decreased the cellular expressions of JAK2 and STAT3. CONCLUSION The anti-inflammatory activity of Eurycoma longifolia is attributed primarily to its active ingredients bitter lignin and alkaloids, which may regulate the JAK/STAT3 signaling pathway by targeting JAK2 and STAT3.
Animals ; Mice ; Network Pharmacology ; Eurycoma ; Lipopolysaccharides ; Molecular Docking Simulation ; Tandem Mass Spectrometry ; Anti-Inflammatory Agents/pharmacology* ; Ethanol ; Plant Extracts/pharmacology*

A sensitive and efficient method was established and validated for qualitative and quantitative analysis of total alkaloids from the extract of Eurycoma longifolia by high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(HPLC-Q-TOF-MS) combined with ultra-performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry(UPLC-QQQ-MS/MS). The HPLC-Q-TOF-MS conditions are as follows: Welch Ultimate XB-C_(18) column(4.6 mm×250 mm, 5 μm) with acetonitrile(containing 0.1% formic acid)-0.1% formic acid in water as mobile phase for gradient elution. The UPLC-QQQ-MS/MS conditions are as below: Agilent Eclipse Plus C_(18) column(2.1 mm×50 mm, 1.8 μm) with acetonitrile(containing 0.1% formic acid) and 0.1% formic acid in water as mobile phase for gradient elution. MS data were collected by electrospray ionization in positive ion mode. According to the comparison with reference standards and the accurate masses of molecules, a total of 17 alkaloids in E. longifolia extract were identified by HPLC-Q-TOF-MS. The UPLC-QQQ-MS/MS quantitative analysis result of 3 alkaloids showed that the linear ranges of them were good(r≥0.999 7) and the overall recoveries ranged from 108.8%-110.2%, with RSDs of 2.9%-5.3%. The method is accurate, reliable, and efficient, which can comprehensively reflect the constituents and content of alkaloids in E. longifolia. The result can serve as a reference for further elucidating its therapeutic material basis and quality control.
Alkaloids ; Chromatography, High Pressure Liquid ; Chromatography, Liquid ; Eurycoma ; Tandem Mass Spectrometry

In this paper,metabolomics and network pharmacology were used to investigate the bioactive components of Harrisonia perforata and their possible mechanisms of action. Metabolites in the flowers,fruits,branches,leaves and stalks of H. perforata were analyzed by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry. Meanwhile,multiple statistical analysis methods including principal component analysis( PCA) and orthogonal partial least squares discriminant analysis( OPLS-DA)were applied to screen and identify differential compounds. With metabolomics method,9 differential compounds were preliminarily identified from leaves and other non-traditional medicinal parts. Subsequently,these compounds were explored by using network pharmacology. With gastrointestinal absorption and drug-likeness as limiting conditions,they were imported into the Swiss ADME,from which 7 compounds with potential medicinal activity were obtained. Then,their targets were predicted by PharmMapper,with Human Protein Targets Only and Normalized Fit Score>0. 9 set as limiting conditions,and 60 standardized potential targets were identified with Uniprot. KEGG( Kyoto encyclopedia of genes and genomes) pathway data was obtained using metascape and the " potential active ingredients-target-pathway" network was constructed with Cytoscape 3. 7. 2. The enrichment analysis of KEGG demonstrated that the 60 targets were enriched in 78 signaling pathways( min overlap: 3,P value cutoff: 0. 01,min enrichment: 1. 5),many of which are related to anti-bacteria,anti-inflammation and anti-virus,such as IL-17 signaling pathway,RIG-I-like receptor signaling pathway and NOD-like receptor signaling pathway. Finally,depending on the clinical activity of H. perforata,the relevant signaling pathways were analyzed through experimental data and literature. Dehydroconiferyl alcohol was reported to have the anti-inflammatory effect and perforamone D to possess the antimycobacterial activity. The KEGG pathway enrichment analysis showed that dehydroconiferyl alcohol could act on the Alzheimer's disease( AD) signaling pathway by targeting CDK5 R1 and BACE1. ACh E inhibitor is the most promising drug to treat AD,while dehydroconiferyl alcohol has been proved to inhibit ACh E according to literature. The experimental results revealed that the extract of leaves of H. perforata can effectively inhibit the growth of Staphylococcus aureus. These are consistent with the enrichment analysis results of KEGG. This study explored the bioactive components and pharmacodynamics of the leaves of the H. perforata,laying a theoretical foundation for its in-depth development and rational application.
Amyloid Precursor Protein Secretases ; Aspartic Acid Endopeptidases ; Drugs, Chinese Herbal/pharmacology* ; Humans ; Metabolomics ; Simaroubaceae

Brucea javanica oil emulsion (BJOE) has been used to treat tumor in China for more than 40 years. However, its components and effectiveness in the treatment of acute lymphocytic leukemia (ALL) and its mechanism of anti-cancer activity remain unknown. In the current study, high-performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD) was used to analyze the components of BJOE. Then, the anti-leukemia effects of BJOE were examined both in vitro and in vivo using ALL Jurkat cells and the p388 mouse leukemia transplant model, respectively. The primary ALL leukemia cells were also used to confirm the anti-leukemia effects of BJOE. The apoptotic-related results indicated that BJOE induced apoptosis in Jurkat cells and were suggestive of intrinsic apoptotic induction. Moreover, BJOE inhibited Akt (protein kinase B) activation and upregulated its downstream targets p53 and FoxO1 (forkhead box gene, group O-1) to initiate apoptosis. The activation of GSK3β was also involved. Our findings demonstrate that BJOE has anti-leukemia effects on ALL cells and can induce apoptosis in Jurkat cells through the phosphoinositide3-kinase (PI3K) /Akt signaling pathway.
Animals ; Apoptosis ; Brucea/chemistry* ; Glycogen Synthase Kinase 3 ; Humans ; Jurkat Cells ; Mice ; Phosphatidylinositol 3-Kinases/genetics* ; Plant Oils/pharmacology* ; Precursor Cell Lymphoblastic Leukemia-Lymphoma/drug therapy* ; Proto-Oncogene Proteins c-akt/genetics* ; Seeds/chemistry* ; Signal Transduction

Angiogenesis is a crucial process in the development of inflammatory diseases, including cancer, psoriasis and rheumatoid arthritis. Recently, several alkaloids from Picrasma quassioides had been screened for angiogenic activity in the zebrafish model, and the results indicated that 1-methoxycarbony-β-carboline (MCC) could effectively inhibit blood vessel formation. In this study, we further confirmed that MCC can inhibit, in a concentration-dependent manner, the viability, migration, invasion, and tube formation of human umbilical vein endothelial cells (HUVECs) in vitro, as well as the regenerative vascular outgrowth of zebrafish caudal fin in vivo. In the zebrafish xenograft assay, MCC inhibited the growth of tumor masses and the metastatic transplanted DU145 tumor cells. The proteome profile array of the MCC-treated HUVECs showed that MCC could down-regulate several angiogenesis-related self-secreted proteins, including ANG, EGF, bFGF, GRO, IGF-1, PLG and MMP-1. In addition, the expression of two key membrane receptor proteins in angiogenesis, TIE-2 and uPAR, were also down-regulated after MCC treatment. Taken together, these results shed light on the potential therapeutic application of MCC as a potent natural angiogenesis inhibitor via multiple molecular targets.
Angiogenesis Inhibitors ; chemistry ; pharmacology ; Animals ; Carbolines ; chemistry ; pharmacology ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Epidermal Growth Factor ; genetics ; metabolism ; Fibroblast Growth Factors ; genetics ; metabolism ; Human Umbilical Vein Endothelial Cells ; cytology ; drug effects ; metabolism ; Humans ; Insulin-Like Growth Factor I ; genetics ; metabolism ; Neovascularization, Physiologic ; drug effects ; Picrasma ; chemistry ; Plant Extracts ; chemistry ; pharmacology ; Receptor, TIE-2 ; genetics ; metabolism ; Zebrafish ; embryology

In the present study, a gastric retention floating system for Brucea javanica oil, composed of alginate and carrageenan, was prepared using ionotropic gelation. Parameters for floatability, drug load, encapsulation efficiency, bead morphology, in vitro release, and in vivo gastric retention were evaluated. The optimized formulation via Box-Behnken design consisted of 1.7% alginate (W/V), 1.02% carrageenan (W/V), 1.4% CaCO (W/V), and a gelling bath of pH 0.8. The alginate-carrageenan-Brucea javanica oil beads had a porous structure and exhibited up to 24 h of in vitro floatability with a load capacity of 45%-55% and an encapsulation efficiency of 70%-80%. A 6-h sustained release was observed in vitro. The beads had a prolonged gastric retention (> 60% at 6 h) in fasted rats, compared to non-floating beads (15% at 6 h), as measured by gamma scintigraphy with single-photon emission tomography/computed tomography (SPET/CT). In conclusion, the alginate-carrageenan-Brucea javanica oil system showed enhanced oil encapsulation efficiency, excellent floating and gastric retention abilities, and a favorable release behavior.
Alginates ; chemistry ; Animals ; Biological Availability ; Brucea ; chemistry ; Carrageenan ; chemistry ; Delayed-Action Preparations ; administration & dosage ; chemistry ; pharmacokinetics ; Drug Carriers ; chemistry ; Drug Delivery Systems ; methods ; Drug Evaluation, Preclinical ; Gastric Mucosa ; metabolism ; Glucuronic Acid ; chemistry ; Hexuronic Acids ; chemistry ; Microspheres ; Plant Oils ; administration & dosage ; chemistry ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley

To investigate the effect of Brucea javanica oil emulsion on proliferation, migration and autophagy of non-small cell lung cancer A549 cells.
 Methods: First, A549 cells were divided into a control group and a low, medium or high dose of Brucea javanica oil emulsion groups (0, 2.5, 5.0 or 10.0 mg/mL); then, the cells were divided into a 3-MA+Brucea javanica oil emulsion group and a Brucea javanica oil emulsion group in the presence or absence of 3-methyl adenine (3-MA). Cell counting kit-8 (CCK-8) and clone formation assay were used to detect cell proliferation, while the wound scratch and Transwell assay were used to measure cell migration. Cell immunofluorescence and Western blot were used to analyze autophagy.
 Results: Compared with the control group, the numbers of cell proliferation and colony-formation, the relative cell migration rate and numbers of trans-membrane cells were reduced in a dose-dependent manner in the Brucea javanica oil emulsion groups (all P<0.05). Meanwhile, compared with the control group, the aggregation of microtubule associated protein 1 light chain3 (LC3) green fluorescence and the LC3-II/LC3-I ratios were increased, while p62 level was decreased (all P<0.05) in the high dose group. Compared with the Brucea javanica oil emulsion group (5.0 mg/mL), the cell proliferation, numbers of cell clone formation, cell migration rate and numbers of Transwell transmembrane cells were increased in the 3-MA+Brucea javanica oil emulsion group (all P<0.05).
 Conclusion: Brucea javanica oil emulsion can promote the autophagy of non-small cell lung cancer A549 cells and inhibit the cell proliferation and migration.
A549 Cells ; Autophagy ; drug effects ; Brucea ; chemistry ; Carcinoma, Non-Small-Cell Lung ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Humans ; Lung Neoplasms ; Plant Oils ; pharmacology

Brucea javanica, a Chinese herbal medicine, combined with conventional anticancer modalities, has been widely used for treatment of various cancers. Based on researches over the last decades, authors briefly summarized its active constituents, molecular mechanisms and clinical application for cancer treatment.
Antineoplastic Agents, Phytogenic ; therapeutic use ; Apoptosis ; drug effects ; Biomedical Research ; methods ; trends ; Brucea ; chemistry ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Neoplasms ; drug therapy ; pathology ; Phytotherapy