UPLC-Q-TOF-MS/MS combined with network pharmacology for exploring antiinflammatory mechanism of Eurycoma longifolia / 南方医科大学学报

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.

Exploring dynamic biomedical algorithm of Eurycoma longifolia Jack and its bioactive phytochemicals: A review of pharmacokinetic and pharmacodynamic implications and future prospects

Eurycoma longifolia Jack (E. longifolia) is a well-recognized traditional herbal medicine that offers a wide dynamic range of biomedical applications including anti-osteoporotic, anticancer, anti-proliferative, anti-malarial, antimicrobial, antioxidant, aphrodisiac, anti-inflammatory, anxiolytic, anti-diabetic, anti-rheumatism and anti-ulcer properties. This review aims to overview the pharmacokinetic and a pharmacodynamic algorithm of E. longifolia and its bioactive components. Analysis of pharmacokinetic profile revealed that E. longifolia exhibit higher bioavailability, high volume of distribution, slow elimination rate, and does not show inhibitory effects on cytochrome P450 isoenzymes. E. longifolia has been used, alone or in combination with other pharmacological agents, in the form of crude extracts, standard extracts, or decoctions of different plant parts (i.e., herbs, shrubs, stem, leaves, and roots) for the treatment of various ailments in animals and humans. Among various bioactive constituents, eurycomanone has been found to be the most remarkable, super-stable, versatile, and most potent phytochemical (isolated or extracted from root extracts) against various types of animals and human diseases. Based on its well-established pharmacokinetic and pharmacodynamic profiles, we suggested that E. longifolia can be a well-accepted complementary and alternative medicine for the treatment of different types of human ailments.

Extraction and purification process of the active ingredient eurycomanone from Eurycoma longifolia Jack / 中国药学杂志

OBJECTIVE: To optimize the extraction and purification process of eurycomanone (EN) from Eurycoma longifolia Jack. METHODS: Using single factor test and orthogonal test to screen the best conditions of ethanol concentration, extraction time, amount of solvent and extraction times with the index of transfer rate of EN. Resin model was screened by static adsorption and desorption test, purification process of EN was investigated by single factor test and orthogonal test. RESULTS: The optimal condition of the extracting was 20-fold water, 2 times with the first 2 and 1 h for the second time. Optimal purification process of HPD100 macroporous resin was 70% amount of saturated adsorption sample, 0.25 g·mL-1 of sample liquid concentration, 3BV·h-1 of sample flow rate, eluting using the 30% ethanol, 3 BV·h-1 of elution velocity. CONCLUSION: The optimized process is simple, stable and repeatable, which is suitable for industrial production.

In Vitro Antibacterial Activity of Eurycoma Longifolia Jack (Tongkat Ali) Root Extract

Currently, researchers are aiming to explore herbal plants to replace synthetic drugs because herbal plants contain high active compounds and fewer side effects. Our study was done to determine the antibacterial activity of Eurycoma longifolia Jack (E. longifolia) root using ethanol based extract. Methods: Five types of pathogenic bacterial strains were used; Gram-positive (Staphylococcus aureus and Bacillus cereus) and Gram-negative (Escherichia coli, Salmonella typhi and Pseudomonas aeruginosa). Disc diffusion assay and Minimum Inhibitory Concentration (MIC) tests were used to determine the inhibition zone and turbidity of suspension which reflects the antibacterial activity of the extract. Results: The ethanolic extract of E. longifolia Jack root extract showed positive results against Gram-positive bacteria (S. aureus and B. cereus) and Gramnegative (S. typhi). B.cereus and S.typhi showed inhibition zone values of 11.76mm and 14.33mm at the extract concentration of 150mg/ml that were higher than the positive control values (9.00, 12.67mm) respectively. However, E. coli and P. aeruginosa did not show any inhibition by the ethanol-based extract. Conclusion: From the results we can conclude that E.Longifolia root extract possesses antibacterial activity that can be further explored to produce new medicinal products.

Effect of Long-Term Use of Eurycoma Longifolia Jack on Histopathological Changes in the Liver in Rats

Presently, the use of Eurycoma longifolia Jack (ELJ) (Tongkat Ali) has increased dramatically in Southeast Asia especially Malaysia where it is widely used as aphrodisiac and anti-malarial agent. Interestingly, its consumption has become popular in daily life as beverage to enhance energy and stamina especially among males. However, its effect on the safety of vital organs of the body has not been studied adequately. Hence, the main objective of this study was to determine whether or not long-term use of ELJ any has side effects on the liver in rats. Methods: Three different concentrations of aqueous extract of ELJ were prepared and dissolved in distilled water. A total of 32 Sprague-Dawley male rats were used and randomly divided into three test groups and control. The test groups were given different doses (low dose 250 mg/kg bw, medium dose 500mg/kg bw and high dose 1000 mg/kg bw) of aqueous extract of ELJ, respectively. Control group was given distilled water alone. Doses were given orally and daily for 5 weeks. After 5 weeks, animals were sacrificed; whole liver tissues were obtained, fixed in 10 percent formaldehyde overnight for histological examination. Result: Histological observations showed mild to moderate degrees of hemorrhage, hepatocytes degeneration and severe fatty changes in liver tissue of the test groups treated with ELJ as compared to control. Conclusion: In conclusion, the long-term daily consumption of ELJ in large quantity as beverage may cause fatty changes, hemorrhage and hepatocytes degeneration in the liver tissue.

Decreased expression of caspase3 in penis and prostate tissues of rat after the treatment with buceng (Pimpinella alpina Molk & Euricoma longifolia Jack).

Background: Buceng {combination of pasak bumi (Eurycoma longifolia Jack) and purwoceng (Pimpinella alpine Molk)} has been proven to increase testosterone (Te) level and decrease apoptosis. Unfortunately, there is no evidence whether these effects are mediated by the declining of caspase3. Objective of this study was to evaluate whether buceng could decrease the expression of caspase3 of penis and prostate cells in Sprague Dawley male rats. Methods: Twenty four Sprague Dawley male rats weighing 300 g (90 days old) were randomly assigned into 4 groups of 6 male rats. Group A, rats were castrated and received buceng 50 mg. Group B, rats were not castrated, sacrifices as positive control. Group C, rats were castrated and given 2 mL aquadest as negative control. Group D, rats were castrated and got of 6.75 mg mesterolone, dissolved in 2 mL water. MANOVA statistical analysis was adopted to examine the difference expression of caspase3 in all groups. The comparison of caspase3 expression between two groups exhibiting difference values were evaluated by Post Hoc test. Results: MANOVA revealed statistically significant differences in the expression of caspase3 of penis and prostate tissues among the four groups. Post Hoct test also indicated that expression of caspase3 in group A (buceng) (33.56; 35.83) was significantly lower compared to group C (negative control) (54.33; 60.07) and group D (mesterolone) (51.91;56.21), p = 0.000, and higher compared than group B or normal rats (29.40; 27.72), but statistically not significant (p = 0.826). Conclusion: The treatment of 50 mg buceng/day for 30 consecutive days could decrease caspase3 expression in penis and prostate cells.

The effect of eurycoma longifolia jack on spermatogenesis in estrogen-treated rats

Introduction: There is little data concerning the ability of Eurycoma longifolia Jack (EL) to reverse the inhibitory effects of estrogen on testosterone production and spermatogenesis. The aim of the present study was to determine the effect of EL on testicular histology and sperm count in estrogen-treated male rats. Methods: Adult male Sprague-Dawley rats weighing 200-250 g were divided into four groups of six rats each. Group A (control) was given solvent in the same manner as the treated groups were given EL. Group B was treated with EL (8 mg/kg body weight) orally. Group C was treated with estradiol (E2) (intramuscular dose of 500 ìg/kg body weight), and group D received a combined treatment of oral EL and intramuscular E2. After fourteen consecutive days of treatment, rats from all groups were sacrificed and subjected to spermatogenic and epididymal sperm cell counts. Results: The spermatogenic cell count in the E2-treated group was significantly decreased as compared to the control (p < 0.05) and EL+E2-treated groups (p < 0.05). A similar finding was found for the epididymal sperm count; the E2-treated group had a significant decrease in the count compared to the control (p < 0.05) and EL+E2-treated groups (p < 0.05). Rats that were treated with EL alone exhibited significantly higher sperm counts and sperm motility when compared to the control group (p < 0.05). Conclusions: EL extract acts as a potential agent for reversing the effects of estrogen by increasing spermatogenesis and sperm counts in rats after fourteen consecutive days of treatment.

Antiplasmodial effects of Brucea javanica (L.) Merr. and Eurycoma longifolia Jack extracts and their combination with chloroquine and quinine on Plasmodium falciparum in culture

Fruits of <I>Brucea javanica</I> (L.) Merr. (“Ratchadad” in Thai) and roots of <I>Eurycoma longifolia</I> Jack (“Plalaipeag” in Thai) are used as traditional medicines for the treatment of malarial fever. Ethanol, methanol, ethyl acetate, ethyl alcohol and aqueous extracts were tested against the multidrug-resistant <I>Plasmodium falciparum</I> strain K1. Ethanol and methanol-ethanol extracts, together with methanol residue, from fruits of <I>B. javanica</I> (L.) Merr. showed the highest antiplasmodial activities with IC<SUB>50</SUB> values of 0.5 ± 0.3, 0.3 ± 0.1 and 0.3 ± 0.05 Μg⁄mL, respectively, comparable to the IC<SUB>50</SUB> values of chloroquine (0.17 ± 0.02 Μg⁄mL) and quinine (0.3 ± 0.1 Μg⁄mL). Similarly, ethanol and methanol-ethanol extracts of roots of <I>E. longifolia</I> Jack showed higher activities than those of the other solvent extracts, but their activities were about 10-fold lower than those of extracts from <I>B. javanica</I> (L.) Merr. fruit. In drug combination tests, <I>B. javanica</I> (L.) Merr. and <I>E. longifolia</I> Jack extracts did not appear to antagonize antiplasmodial activity of chloroquine and quinine. Not only well-known quassinoid glycosides but also coumarins and flavonoids identified by thin-layer chromatography in ethanol and methanol-ethanol extracts and in methanol residue of <I>B. javanica</I> (L.) Merr fruit and <I>E. longifolia</I> roots may be responsible for the antimalarial activity. Taken together, our extraction conditions provided extracts containing novel active compounds that did not antagonize the inhibitory effects of the two widely used antimalarials. This finding could lend support to the future discovery of active antimalaria compounds of <I>Brucea javanica</I> (L.) Merr. and <I>Eurycoma longifolia</I> Jack as drugs for the treatment of malaria that could be employed as drug combinations in order to delay the onset of parasite drug resistance.