Effect in white shrimp Litopenaeus vannamei of Eleutherine bulbosa (Mill.) Urb. Powder on immune genes expression and resistance against Vibrio parahaemolyticus infection.

This study investigated the influence of Eleutherine bulbosa (Mill.) Urb. on the immune responses, bacterial population in the intestines, and resistance of white shrimp, Litopenaeus vannamei, against infection with Vibrio parahaemolyticus. Shrimp were fed with three dosages of powder, at 6.25 g kg-1 (P6.25), 12.5 g kg-1 (P12.5), and 25 g kg-1 (P25). One dosage of the crude extract was provided, 1.25 g kg-1 (E1.25), and the controls without administration of E. bulbosa consisted of a positive control (PC) and a negative control (NC). Feed supplementation was carried out for 30 days; then shrimp from all treatments were challenged by intramuscular injection with V. parahaemolyticus (106 cfu/mL), except for the NC. The results showed that supplementation with the powder and extract of E. bulbosa for 30 days resulted in significantly higher (P < 0.05) immune responses (total hemocyte count (THC), phenoloxidase activity (PO), respiratory bursts (RBs)), gene expression (prophenoloxidase (proPO), lipopolysaccharide- and ß-1,3-glucan-binding protein (LGBP)), and total bacterial count (TBC) compared to PC/NC. In post challenge testing, there were significantly higher levels for THC, PO, RBs, proPO, LGBP, and PE (peroxinetin), and the treatments were able to suppress V. parahaemolyticus in the intestines, hepatopancreas, and muscles and to reduce damage to the muscles and hepatopancreas. The survival rate with P12.5 was significantly higher compared to the other treatments. It was concluded that the shrimp receiving supplementation with the powder and extract of E. bulbosa had increased immunity and resistance against V. parahaemolyticus infection, with the best dosage being the P12.5 treatment.

Exploring the anticancer potential of Eleutherine bulbosa: A systematic network pharmacology study on lung cancer.

Chemotherapy application in lung cancer patients has several side effects and shows lower effectiveness due to chemoresistance. Although Eleutherine bulbosa (Mill.) Urb. (EBE) elicit anticancer properties, yet the exact profile of its active compounds and lung cancer inhibition mechanisms were not fully understood. This study aimed to identify suggestive compounds from EBE extract and explain the molecular mechanisms of EBE against lung cancer. Identification of the compound from the EBE extract was confirmed using liquid chromatography-tandem mass spectrophotometry (LC-MS/MS). The bioavailability profile of three major metabolites was identified using absorption, distribution, metabolism, excretion, toxicity software. The anticancer molecular mechanism prediction of the drugs was ascertained by network pharmacology using Cytoscape 3.9.1 and the protein-protein interaction network technique with STRING 11.0. Interaction between resveratrol and extracellular growth factor receptor (EGFR) was analyzed using site-specific molecular docking with erlotinib as the control using PyRx Autodock Vina 9.0 and BIOVIA Discovery Studio. A total of 16 active compounds were identified from LC-MS/MS. Only resveratrol showed anticancer properties by its interaction with 13 genes and 6 signaling pathways related to lung cancer. The molecular docking result supports the network pharmacology finding. The binding affinity of resveratrol with EGFR, important receptor in lung cancer, was more negative (-6.9 kcal/mol) than erlotinib (-6.2 kcal/mol) as the control. Evidence suggested that resveratrol in EBE exhibits anticancer effects by modulating lung cancer cell proliferation and apoptosis through EGFR binding.

Phenolic constituents with antibacterial activity from Eleutherine bulbosa.

Eleutherine bulbosa (Mill.) Urb. is a medicinal and edible plant with various benefits for humans and animals. In this work, four new phenolic constituents (1-4), along with six known phenolic compounds (5-10) were obtained from the red bulbs of E. bulbosa. Their structures with absolute configurations were characterized by extensive spectroscopic analysis, combined with HR-ESI-MS and quantum mechanical electronic circular dichroism (ECD). Compounds 1 and 2 are novel homologous and heterodimers, respectively, featuring an unusual spiro ring system. All isolated phenolic constituents were tested for their antibacterial effects. The results revealed four phenolic compounds 1-3 and 7 showed moderate antibacterial activity against Bacillus subtilis, Staphylococcus aureus and Escherichia coli with minimum inhibitory concentration (MIC) values ranging from 15.6 to 250.0 µg/mL.

Effects of Eleutherine bulbosa extract on the myofibrillar protein oxidation and moisture migration of yak meat under oxidation stress.

The influence of Eleutherine bulbosa (EB) extract at various levels (1, 4, 7, 10 or 13 g/kg) on the myofibrillar protein oxidation and moisture migration of yak meat in Fenton oxidation system was investigated. The results showed that inclusion of EB extract in yak meat efficiently inhibited carbonyl formation triggered by hydroxyl radicals. Supplementation of EB extract at 1-10 g/kg manifested more contents of the active sulfhydryl, ε-NH2 groups and α-helix structure, and higher solubility of myofibrillar proteins (MPs), but alleviated the turbidity of MPs. However, adding high level of EB extract (13 g/kg) induced the loss of free amine and α-helix content and resulted in more aggregation of MPs. The SDS-PAGE demonstrated that adding 1-7 g/kg EB extract had an obvious protective effect for myosin heavy chain and actin, whereas 10 or 13 g/kg EB extract led to weakened intensities of protein bands. DSC and LF-NMR analysis revealed that 7 g/kg EB extract had appreciable effects on thermal stabilities of MPs, and improved the hydration of yak meat induced by oxidation, while 13 g/kg EB extract accelerated MP structure destabilization and lowered water retention. Our results suggested that incorporation of low levels of EB extract (1-7 g/kg) effectively retarded the oxidative damage to MPs and EB extract could be a promising natural antioxidant in meat processing.

The Antiproliferative Effect of Chloroform Fraction of Eleutherine bulbosa (Mill.) Urb. on 2D- and 3D-Human Lung Cancer Cells (A549) Model.

Since lung cancer is the leading cause of cancer-related death worldwide, research is being conducted to discover anticancer agents as its treatment. Eleutherine bulbosa, a Dayak folklore medicine, exhibited anticancer effects against several cancer cells; however, its anticancer potency against lung cancer cells has not been explored yet. This study aims to determine the anticancer potency of E. bulbosa bulbs against lung cancer cells (A549) using 2D and 3D culture models, as well as determine its active compounds using gas chromatography-mass spectrometry (GC-MS) analysis. Three fractions of E. bulbosa bulbs, namely chloroform, n-hexane, and ethyl acetate, were tested for cytotoxicity using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium-bromide (MTT) and CellTiter-Glo. The antiproliferative effects of the most cytotoxic fraction against the 2D culture model were determined by a clonogenic survival assay and propidium iodide/Hoechst 33342 double staining, whereas the effects against the 3D culture model were determined by microscopy, flow cytometry, and gene expression analysis. The chloroform fraction is the most cytotoxic against A549 cells than other fractions, and it inhibited colony formation and induced apoptosis of A549 cells. The chloroform fraction also inhibited the growth of the A549 spheroid by suppressing the spheroid size, inducing apoptosis, reducing the proportion of CD44 lung cancer stem cells, causing arrest at the S phase of the cell cycle, and suppressing the expression of the SOX2 and MYC genes. Furthermore, the GC-MS analysis detected 20 active compounds in the chloroform fraction, including the major compounds of eleutherine and isoeleutherine. In conclusion, the chloroform fraction of E. bulbosa bulbs exhibit its antiproliferative effect on 2D and 3D culture models of A549 cells, suggesting it could be a lung cancer chemopreventive agent.

Antiosteoarthritis activities of 70% ethanol extract of Eleutherine bulbosa (mill.) urb. bulb on rats monosodium iodoacetate-induced osteoarthritis.

Background: Osteoarthritis (OA) is a common degenerative joint situation that induces pain and disability in the elderly. Traditionally, Eleutherine bulbosa bulb from Pasuruan, East Java, is used to treat many diseases, also as an anti-inflammatory. Objective: In this research, we employed an in vivo model to examine the effects of 70% ethanol extracts of E. bulbosa (EBE) on the progression and development of OA. Methods: A singular intraarticular injection of Monosodium Iodoacetate (MIA) was used to create the OA model in rats. The progression of OA was observed for three weeks. Furthermore, treatment of EBE at a dose of 6, 12, and 24 mg/200g BW orally for four weeks was conducted to assess the effects on decreasing IL- 1. level, joint swelling, and hyperalgesia. Results: Induction was successful, indicated by a significant difference (P<0.05) in decreasing latency time, increasing joint swelling, and IL-1. level. EBE 24 mg/200 g BW treatment has significantly (P<0.05) reduced IL-1. levels, joint swelling, and response to hyperalgesia. Conclusion: The 70% ethanol extract of E. bulbosa bulb has therapeutic effects on inflammation through reducing IL-1. in experimental MIA-induced osteoarthritis in a rat model. According to this study, EBE may have an effective potential new agent for OA therapy.

Effects of Eleutherine bulbosa (mill.) urb. bulb extract on mice glucocorticoid-induced osteoporosis models.

Background: Low bone mass accompanied by microarchitectural alterations in the bone that cause fragility fractures is known as secondary osteoporosis and occurs when there is an underlying condition or medication present. Eleutherine bulbosa bulb extract has been shown to affect bone because of its content, which can help osteoblast differentiation and inhibit osteoclast differentiation. Objective: This study aimed to assess the effects of 70% ethanol extract of E. bulbosa Bulbs (EBE) from Pasuruan-East Java on blood calcium levels, osteoblast cell count, and bone density of trabecular femur in osteoporosis rats. Methods: Six groups of 30 female Wistar rats were created. There were no test materials offered to the healthy group; the negative group received 0.5% CMC; the positive group received alendronate 0.9 mg/kg BW; and the dose group received 30, 60, and 120 mg/kg BW. Glucocorticoid (Dexamethasone) 0.1015 mg/kg BW/day induction was given to all groups except the healthy group to create osteoporosis rats for approximately four weeks. Then they were given oral therapy for approximately 28 days. Followed by the determination of blood calcium levels, the number of osteoblast cells, and bone density of the rat femur trabecular. Results: The result showed that E. bulbosa bulbs extract could raise blood calcium levels and bone density percentage at doses of 60 and 120 mg/kg BW, as well as raise osteoblast cell levels at doses of 120 mg/kg BW. Conclusions: The findings indicate that E.bulbosa bulb extract is a potential complementary medicine for osteoporosis.

Optimization of microwave-assisted extraction on polyphenol metabolite from Eleutherine bulbosa (Mill.) urb. bulbs using response surface methodology.

Eleutherine bulbosa bulbs, an endemic plant in Indonesia, have enormous potential as raw materials for pharmaceutical products. Therefore, it is necessary to strengthen and develop extraction methods that are easy, rapid, and efficient to enrich targeted secondary metabolites. This study aims to optimize the microwave-assisted extraction (MAE) method conditions for polyphenol metabolite from E. bulbosa bulbs. The MAE method (with different conditions) was applied to extract total polyphenol content (TPC) from E. bulbosa bulbs. TPC values were determined using a 96-well microplate reader spectrophotometry method and Folin-Ciocalteu reagent. The variables of MAE, as an experimental design-independent variable, were involved. The MAE method condition was optimized using response surface methodology (RSM) and Box-Behnken design based on the TPC value. The MAE condition was optimized with 60% ethanol, sample-solvent ratio of 1:10 g/mL, and 50% Watts of microwave power for 10 min. The quadratic regression analysis was achieved to predict the TPC value using the equation: TPC value = 28.63-5.545A +2.211B -0.741C +1.995D - 4.045AB +0.856AC -7.541BC +1.961CD -8.342A2-0.071B2 +1.840C2-1.535D2. For the scale-up confirmation test, a 50-g sample was used to prove the validity of the equation to predict the TPC value, yielding 35.33 ± 2.13 mg gallic acid equivalent/g samples. The optimum of the MAE condition recommended based on the results of RSM analysis can be applied directly to the enrichment of polyphenols metabolite constituent of E. bulbosa easily, cheaply, quickly, and efficiently.

The chemical constituents from the active fractions of Eleutherine bulbosa with their antimicrobial activity.

Six compounds including three new polyketide ones named eleubosas A-C (1-3) were isolated from the active frations of Eleutherine bulbosa. Their structures were elucidated by extensive spectroscopic methods, including NMR, MS and IR spectroscopic analyses data. All the isolates were evaluated against three pathogenic bacteria, Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa, and the results showed that compounds 1 and 2 displayed moderate inhibitory activities against E. coli with MIC values both 12.5 µg/mL, which are consistent with the clinical applications and need further studies.

Phytochemical analysis and antibacterial activities of Eleutherine bulbosa (Mill.) Urb. extract against Vibrio parahaemolyticus

To analyze compounds in Eleutherine bulbosa (E. bulbosa) (Mill.) Urb. extract and to determine its antibacterial capability against Vibrio parahaemolyticus (V. parahaemolyticus). Methods: E. bulbosa bulb extract was preprared using 96％ ethanol by the maceration method. Phytochemical investigation of E. bulbosa extract was analyzed using GC-MS, spectrophotometry and titrimetry methods. The zone of inhibition was identified by the diffusion agar method. The minimum inhibitory concentration and minimum bactericidal concentration were determined using the plate count method. The inhibitory rate against V. parahaemolyticus was determined by the microdilution method. Cellular leakage was evaluated by spectrophotometry and cellular damage was observed by scanning electron microscopy. Results: GC-MS analysis showed the high compound of the E. bulbosa extract was securixanthone E (7-hydroxy-1,2-dimethoxyxanthone). The compound groups also included fatty acid esters, isoquinolines, naphthalenes, and phenolics. The total phenolic content was (2.50 ± 0.00) mg/g, flavonoid (6.61 ± 0.00) mg/g, and tannin (0.03 ± 0.00)％. The greatest zone of inhibition and inhibitory rate were (11.83 ± 0.06) mm and (91.32 ± 2.76)％, respectively, at 10 mg/mL. The minimum inhibitory concentration was 0.156 mg/mL, while the minimum bactericidal concentration was 10 mg/mL. The E. bulbosa extract caused leakage and cellular damage to V. parahaemolyticus. Conclusions: The E. bulbosa extract possesses inhibitory activities against V. parahaemolyticus and causes cellular leakage and damage.