Transcriptomic data reveals the dynamics of terpenoids biosynthetic pathway of fenugreek.

Medicinal plants are rich sources for treating various diseases due their bioactive secondary metabolites. Fenugreek (Trigonella foenum-graecum) is one of the medicinal plants traditionally used in human nutrition and medicine which contains an active substance, called diosgenin, with anticancer properties. Biosynthesis of this important anticancer compound in fenugreek can be enhanced using eliciting agents which involves in manipulation of metabolite and biochemical pathways stimulating defense responses. Methyl jasmonate elicitor was used to increase diosgenin biosynthesis in fenugreek plants. However, the molecular mechanism and gene expression profiles underlying diosgening accumulation remain unexplored. In the current study we performed an extensive analysis of publicly available RNA-sequencing datasets to elucidate the biosynthesis and expression profile of fenugreek plants treated with methyl jasmonate. For this purpose, seven read datasets of methyl jasmonate treated plants were obtained that were covering several post-treatment time points (6-120 h). Transcriptomics analysis revealed upregulation of several key genes involved in diosgenein biosynthetic pathway including Squalene synthase (SQS) as the first committed step in diosgenin biosynthesis as well as Squalene Epoxidase (SEP) and Cycloartenol Synthase (CAS) upon methyl jasmonate application. Bioinformatics analysis, including gene ontology enrichment and pathway analysis, further supported the involvement of these genes in diosgenin biosynthesis. The bioinformatics analysis led to a comprehensive validation, with expression profiling across three different fenugreek populations treated with the same methyl jasmonate application. Initially, key genes like SQS, SEP, and CAS showed upregulation, followed by later upregulation of Δ24, suggesting dynamic pathway regulation. Real-time PCR confirmed consistent upregulation of SQS and SEP, peaking at 72 h. Additionally, candidate genes Δ24 and SMT1 highlighted roles in directing metabolic flux towards diosgenin biosynthesis. This integrated approach validates the bioinformatics findings and elucidates fenugreek's molecular response to methyl jasmonate elicitation, offering insights for enhancing diosgenin yield. The assembled transcripts and gene expression profiles are deposited in the Zenodo open repository at https://doi.org/10.5281/zenodo.8155183 .

Effect of salinity stress and surfactant treatment with zinc and boron on morpho-physiological and biochemical indices of fenugreek (Trigonella foenum-graecum).

Micronutrient application has a crucial role in mitigating salinity stress in crop plants. This study was carried out to investigate the effect of zinc (Zn) and boron (B) as foliar applications on fenugreek growth and physiology under salt stress (0 and 120 mM). After 35 days of salt treatments, three levels of zinc (0, 50, and 100 ppm) and two levels of boron (0 and 2 ppm) were applied as a foliar application. Salinity significantly reduced root length (72.7%) and shoot length (33.9%), plant height (36%), leaf area (37%), root fresh weight (48%) and shoot fresh weight (75%), root dry weight (80%) and shoot dry weight (67%), photosynthetic pigments (78%), number of branches (50%), and seeds per pod (56%). Fenugreek's growth and physiology were improved by foliar spray of zinc and boron, which increased the length of the shoot (6%) and root length (2%), fresh root weight (18%), and dry root weight (8%), and chlorophyll a (1%), chlorophyll b (25%), total soluble protein content (3%), shoot calcium (9%) and potassium (5%) contents by significantly decreasing sodium ion (11%) content. Moreover, 100 ppm of Zn and 2 ppm of B enhanced the growth and physiology of fenugreek by reducing the effect of salt stress. Overall, boron and zinc foliar spray is suggested for improvement in fenugreek growth under salinity stress.

Effect of hydroalcoholic extract of Trigonella foenum-graecum leaves on wound healing in type 1 diabetic rats.

Diabetes describes a group of metabolic disorders characterised by increased blood glucose concentration. People living with diabetes have a higher risk of morbidity and mortality than the general population. In 2015 it was estimated that there were 415 million (uncertainty interval: 340-536 million) people with diabetes aged 20-79 years, and 5.0 million deaths attributable to diabetes. When diabetic patients develop an ulcer, they become at high risk for major complications, including infection and amputation. The pathophysiologic relationship between diabetes and impaired healing is complex. Vascular, neuropathic, immune function, and biochemical abnormalities each contribute to the altered tissue repair. The use of herbal medicine has increased and attracted the attention of many researchers all over the world. In this study, we have evaluated the effect of 500mg/kg hydroalcoholic extract of Trigonella foenum-graecum leaves (TFG-E) on wound healing in diabetic rats using a full-thickness cutaneous incisional wound model. Wounds of treated animals showed better tensiometric indices, accelerated wound contraction, faster re-epithelialisation, improved neovascularisation, better modulation of fibroblasts and macrophage presence in the wound bed and moderate collagen formation.

Identification of some bioactive compounds from Trignonella foenumgraecum as possible inhibitors of PPARϒ for diabetes treatment through molecular docking studies, pharmacophore modelling and ADMET profiling: An in-silico study.

Oral antidiabetic agents including the peroxisome proliferator-activated receptor gamma (PPARγ) agonists are available for the clinical management of diabetes mellitus (DM) but most are characterized by many adverse effects. In this study, we explore the antidiabetic properties of phytoconstituents from Trigonellafeonumgraecum (Fabaceae) as potential agonist of PPARγ; using in silico molecular docking, molecular mechanics generalized surface area (MM/GBSA)free binding energy prediction, Pharmacophore modeling experiment, and Pharmacokinetic/ toxicity analysis. One hundred and forty (140) compounds derived from Trigonellafeonumgraecum were screened by molecular docking against protein target PDB 3VI8. Results obtained from binding affinity (BA) and that of binding free energy (BFE) revealed five 5 compounds; arachidonic acid (CID_10467, BA -10.029, BFE -58.9), isoquercetin (CID_5280804, BA -9.507kcal/mol, BFE -56.33), rutin (CID_5280805, BA -9.463kcal/mol, BFE -56.33), quercetin (CID_10121947, BA -11.945kcal/mol, BFE -45.89) and (2S)-2-[[4-methoxy-3-[(pyrene-1-carbonylamino)methyl]phenyl]methyl]butanoic acid (CID_25112371, BA -10.679kcal/mol, BFE -45.73); and were superior to the standard; Rosiglitazone with a docking score of -7.672. Hydrogen bonding was notable in the protein-ligand complex interaction, with hydrophobic bond, polar bond and pipi stacking also observed. Their Pharmacokinetic/ toxicity profile showed varying druggable characteristics, but; arachidonic acid had the most favorable characteristics. These compounds are potential agonists of PPARγ and are considered as antidiabetic agents after successful experimental validation.

Evaluation of oxidative stress and genotoxicity of 900 MHz electromagnetic radiations using Trigonella foenum-graecum test system.

Unprecedented growth in the communication sector and expanded usage of the number of wireless devices in the past few decades have resulted in a tremendous increase in emissions of non-ionizing electromagnetic radiations (EMRs) in the environment. The widespread EMRs have induced many significant changes in biological systems leading to oxidative stress as well as DNA damage. Considering this, the present study was planned to study the effects of EMRs at 900 MHz frequency with the power density of 10.0 dBm (0.01 W) at variable exposure periods (0.5 h, 1 h, 2 h, 4 h, and 8 h per day for 7 days) on percentage germination, morphological characteristics, protein content, lipid peroxidation in terms of malondialdehyde content (MDA), and antioxidant defense system of Trigonella foenum-graecum test system. The genotoxicity was also evaluated using similar conditions. It was observed that EMRs significantly decreased the germination percentage at an exposure time of 4 h and 8 h. Fresh weight and dry weight of root and shoot did not show significant variations, while the root and shoot length have shown significant variations for 4 h and 8 h exposure period. Further, EMRs enhanced MDA indicating lipid peroxidation. In response to exposure of EMRs, there was a significant up-regulation in the activities of enzymes such as ascorbate peroxidase (APX), superoxide dismutase (SOD), glutathione-S-transferase (GST), guaiacol peroxidase (POD), and glutathione reductase (GR) in the roots and shoots of Trigonella-foenum graecum. The genotoxicity study showed the induction of chromosomal aberrations in root tip cells of the Trigonella foenum-graecum test system. The present study revealed the induction of oxidative stress and genotoxicity of EMRs exposure in the test system.

Ameliorating effect of Trigonella foenum-graecum L. (fenugreek) extract tablet on exhaustive exercise-induced fatigue in rats by suppressing mitophagy in skeletal muscle.

OBJECTIVE: Trigonella foenum-graecum L. (fenugreek) is widely used as a leafy vegetable and spice in China and North Africa. Recent studies have reported that fenugreek can reduce fatigue; however, its antifatigue mechanism remains unclear. Therefore, this study aimed to investigate the potential antifatigue effects of fenugreek extract (FE) on mitophagy and the underlying mechanisms. MATERIALS AND METHODS: We evaluated the potential effects of FE tablet on an exhaustive exercise-induced fatigue (EEF) rat model. Oxidative stress indicators and fatigue biomarkers in the serum and skeletal muscle were detected. Mitophagy and mitochondrial morphology were observed using transmission electron microscopy. The expression levels of mitochondrial autophagy-related proteins were detected using western blot and immunofluorescence. RESULTS: Compared with the model group, FE enhanced the activities of the antioxidant enzymes superoxide dismutase and glutathione peroxidase as well as total antioxidant capacity; however, it decreased the level of malondialdehyde in the serum and skeletal muscle after a 7-day treatment. Moreover, certain indicators of mitochondrial function, such as reactive oxygen species levels, ATP levels, cellular and mitochondrial Ca2+ levels, and ATPase activity, were significantly improved in the FE group compared with the model group. Finally, we found that mitophagy was induced by exhaustive exercise and inhibited by FE. Regarding mitochondrial autophagy-related proteins, the expression levels of LC3B, FUNDC1, PGAM5, PARKIN, and PINK1 in the skeletal muscle tissue were increased in the EEF group compared with the control group. After administration of FE and a positive control drug, a significant reversal in the expression of the above-mentioned proteins was noted. CONCLUSIONS: Our findings demonstrate that FE exerted antifatigue effects in the EEF rat model by regulating the mitophagy-related FUNDC1/LC3B signaling pathway rather than the PINK1/PARKIN signaling pathway.

Exploring the medicinally important secondary metabolites landscape through the lens of transcriptome data in fenugreek (Trigonella foenum graecum L.).

Fenugreek (Trigonella foenum-graecum L.) is a self-pollinated leguminous crop belonging to the Fabaceae family. It is a multipurpose crop used as herb, spice, vegetable and forage. It is a traditional medicinal plant in India attributed with several nutritional and medicinal properties including antidiabetic and anticancer. We have performed a combined transcriptome assembly from RNA sequencing data derived from leaf, stem and root tissues. Around 209,831 transcripts were deciphered from the assembly of 92% completeness and an N50 of 1382 bases. Whilst secondary metabolites of medicinal value, such as trigonelline, diosgenin, 4-hydroxyisoleucine and quercetin, are distributed in several tissues, we report transcripts that bear sequence signatures of enzymes involved in the biosynthesis of such metabolites and are highly expressed in leaves, stem and roots. One of the antidiabetic alkaloid, trigonelline and its biosynthesising enzyme, is highly abundant in leaves. These findings are of value to nutritional and the pharmaceutical industry.

[Preparation and characterization of fenugreek leaf flavonoids and their protective effects against oxidative damage to hepatocytes].

The present study investigated the main components of fenugreek(Trigonella foenum-graecum L.) leaf flavonoids(FLFs) and their antioxidant activity. FLFs were prepared and enriched by solvent extraction, and the flavonoids were characterized by high-performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry(HPLC-Q-TOF-MS/MS). The protective effect of FLFs against H_2O_2-induced stress damage to L02 hepatocytes was also investigated. Firstly, the cell viability was measured by MTT assay. The oxidative stress injury model was induced by H_2O_2 in L02 cells. The release of lactate dehydrogenase(LDH), the content of reduced glutathione(GSH) and malondialdehyde(MDA), and the activities of superoxide dismutase(SOD) and catalase(CAT) were measured by assay kits. Hoechst fluorescence staining was performed to observe the cell apoptosis. The expression levels of c-Jun N-terminal kinase(JNK), extracellular signal-regulated kinase 1/2(ERK1/2), nuclear factor erythroid-2 related factor 2(Nrf2), heme oxygenase 1(HO-1), and their phosphorylated proteins were detected by Western blot. Based on the MS fragment ion information and data in databases, FLFs contained eight flavonoids with quercetin and kaempferol as the main aglycons. The cell viabi-lity assay revealed that as compared with the conditions in the H_2O_2 treatment group, 3.125-25 µg·mL~(-1) FLFs could increase the viability of L02 cells, reduce LDH release and MDA content in a dose-dependent manner, potentiate the activities of SOD, CAT, and GSH, decrease the phosphorylation of JNK and ERK1/2 proteins, and up-regulate the expression of Nrf2 and HO-1. The results of fluorescence staining showed that the nucleus of the H_2O_2 treatment group showed concentrated and dense strong blue fluorescence, while the blue fluorescence intensity of the FLFs group decreased significantly. FLFs showed a protective effect against H_2O_2-induced oxidative damage in L02 cells, and the underlying mechanism is associated with the enhancement of cell capability in clearing oxygen free radicals and the inhibition of apoptosis by the activation of the MAPKs/Nrf2/HO-1 signaling pathway. The antioxidant effect of fenugreek leaf is related to its rich flavonoids.

22R- but not 22S-hydroxycholesterol is recruited for diosgenin biosynthesis.

Diosgenin is an important compound in the pharmaceutical industry and it is biosynthesized in several eudicot and monocot species, herein represented by fenugreek (a eudicot), and Dioscorea zingiberensis (a monocot). Formation of diosgenin can be achieved by the early C22,16-oxidations of cholesterol followed by a late C26-oxidation. This study reveals that, in both fenugreek and D. zingiberensis, the early C22,16-oxygenase(s) shows strict 22R-stereospecificity for hydroxylation of the substrates. Evidence against the recently proposed intermediacy of 16S,22S-dihydroxycholesterol in diosgenin biosynthesis was also found. Moreover, in contrast to the eudicot fenugreek, which utilizes a single multifunctional cytochrome P450 (TfCYP90B50) to perform the early C22,16-oxidations, the monocot D. zingiberensis has evolved two separate cytochrome P450 enzymes, with DzCYP90B71 being specific for the 22R-oxidation and DzCYP90G6 for the C16-oxidation. We suggest that the DzCYP90B71/DzCYP90G6 pair represent more broadly conserved catalysts for diosgenin biosynthesis in monocots.

Hepatorenal Protective Effect of Fenugreek Aqueous Extract against Lead Toxicity in Experimental Rats.

In this study, aqueous extract of germinated fenugreek seeds was investigated to assess its therapeutic effect on hepatorenal lead toxicity in experimental rats. After overnight fasting, rats were injected intraperitoneally with 0.5 mL of lead acetate at a dose of 35 mg/kg body weight for five consecutive days. Animals were divided into four groups of ten rats each: normal control; untreated negative control and rats treated with 200 or 400 mg/kg body weight of the aqueous extract. Treatments were performed by intraperitoneal injection of 1mL of the extract once a day for 28 consecutive days. Results showed significant differences between treated and control groups during the whole period of the experiment. This was demonstrated by improving body weight and level of serum total protein, decreasing levels of serum ALT, AST, total bilirubin, blood urea nitrogen, and creatinine. As well, histological analysis revealed a marked reduction in inflammation and structural alterations of liver and kidney organs of fenugreek-treated rats. This hepatoprotective effect can be attributed to the anti-inflammatory, anti-oxidant and regenerative capacity of the high content of the phytochemical constituents in the extract.

Kinetin mitigates Cd-induced damagesto growth, photosynthesis and PS II photochemistry of Trigonella seedlings by up-regulating ascorbate-glutathione cycle.

Cytokinins (CKs) plays a key role in plant adaptation over a range of different stress conditions. Here, we analyze the effects of a cytokinin (i.e., kinetin, KN) on the growth, photosynthesis (rate of O2 evolution), PS II photochemistry and AsA-GSH cycle in Trigonella seedlings grown under cadmium (Cd) stress. Trigonella seeds were sown in soil amended with 0, 3 and 9 mg Cd kg-1 soil, and after 15 days resultant seedlings were sprayed with three doses of KN, i.e.,10 µM (low, KNL), 50 µM (medium, KNM) and 100 µM (high, KNH); subsequent experiments were performed after 15 days of KN application, i.e., 30 days after sowing. Cadmium toxicity induced oxidative damage as shown by decreased seedling growth and photosynthetic pigment production (Chl a, Chl b and Car), rates of O2-evolution, and photochemistry of PS II of Trigonella seedlings, all accompanied by an increase in H2O2 accumulation. Supplementation with doses of KN at KNL and KNM significantly improved the growth and photosynthetic activity by reducing H2O2 accumulation through the up-regulation AsA-GSH cycle. Notably, KNL and KNM doses stimulated the rate of enzyme activities of APX, GR and DHAR, involved in the AsA-GSH cycle thereby efficiently regulates the level of AsA and GSH in Trigonella grown under Cd stress. The study concludes that KN can mitigate the damaging effects of Cd stress on plant growth by maintaining the redox status (>ratios: AsA/DHA and GSH/GSSG) of cells through the regulation of AsA-GSH cycle at 10 and 50 µM KN under Cd stress conditions. At 100 µM KN, the down-regulation of AsA-GSH cycle did not support the growth and PS II activity of the test seedlings.

Cytotoxic activity of standardized extracts, a fraction, and individual secondary metabolites from fenugreek seeds against SKOV-3, HeLa and MOLT-4 cell lines.

CONTEXT: Trigonella foenum-graecum L. (Fabaceae) has many therapeutic properties and anticancer potential. OBJECTIVE: The cytotoxic activities of standardized extracts and a fraction from fenugreek seeds and their compounds (sapogenins, flavone C-glycosides, alkaloid trigonelline) against human cancer SKOV-3, HeLa and MOLT-4 cells were evaluated. MATERIALS AND METHODS: Fenugreek seeds were extracted with 70% methanol (A) or water (B). Furthermore, the seeds were purified with petroleum ether and chloroform and next extracted with methanol to obtain fraction (C). The quantitative analysis of saponins and flavonoids in the extracts was done with HPLC methods. The extracts (5-120 µg/mL) and compounds (1-50 µg/mL) were tested on the cells by MTT assay and RTCA system. The effect of a fraction on ROS production, mitochondrial membrane potential and caspase-3/7 activity in HeLa and SKOV-3 cells was also evaluated by flow cytometry. RESULTS: The strongest cytotoxic activity on cancer cells showed the fraction C (IC50 was 3.91 ± 0.03 for HeLa, 3.97 ± 0.07 for SKOV-3, and 7.75 ± 0.37 for MOLT-4) with the highest content of steroidal saponins (163.18 ± 11.03 µg/mg) and flavone C-glycosides (820.18 ± 0.05 µg/mg). The fraction significantly increased ROS production (up to four times higher than in keratinocytes as control) and caspases activity in the cells. The examined flavonoids did not exhibit the cytotoxic activity in contrast to yamogenin, tigogenin, and diosgenin. CONCLUSIONS: The obtained results complement the data on the cytotoxic activity of Foenugraeci Semen and synergistic effect of flavonoids and saponins complex contained in the plant.

Fenugreek Cultivation with Emphasis on Historical Aspects and its uses in Traditional Medicine and Modern Pharmaceutical Science.

Fenugreek (Trigonella foenum-graecum L.) is a native plant found in the parts of Iran to the North of India, and is presently planted also in other regions of the world. Fenugreek is considered a notable multipurpose medicinal and traditional herb in Iran, India, and China for several centuries. The most important components of fenugreek seeds are protein, neutral detergent fiber, gum, lipids, moisture, ash and starch. Fenugreek seeds and leaves are anti-cholesterolemic, anti-tumor, antiinflammatory, carminative, demulcent, deobstruent, emollient, expectorant, galactogogue, febrifuge, laxative, hypoglycaemic, restorative, parasiticide and uterine tonic and useful in burning sensation. Traditionally, fenugreek seeds being used worldwide are beneficial for bone and muscles, respiratory system, gastro-intestinal system, female reproductive system, cardio-vascular system, endocrinology and hepatic. Fenugreek helps reduce cholesterol, reduce cardiovascular risk, control diabetes, a good consolation for sore throats, a remedy for acid reflux, constipation, colon cancer prevention, appropriate for kidney trouble, skin infection, increase milk production, reduce menstrual discomfort, and reduce menopause symptoms. It is also an appetite suppressant that helps in weight loss. Both modern science and traditional medicine integration with novel technologies and discoveries will secure the cultivation of medicinal herbs and promote sustainability in the long-term and a wide-range.

In Vitro Permeability of Saponins and Sapogenins from Seed Extracts by the Parallel Artificial Membrane Permeability Assay: Effect of in Vitro Gastrointestinal Digestion.

The permeability of saponins and sapogenins from fenugreek and quinoa extracts, as well as dioscin and diosgenin, was evaluated by the parallel artificial membrane permeability assay (PAMPA). The effect of the digestion process on permeability was determined, with previous development of a gastrointestinal process coupled to PAMPA. Saponins from both seeds displayed a moderate-to-poor permeability (>1 × 10-6 cm/s), although the digestion enhanced their permeability values in the order of 10-5 cm/s (p < 0.001). Sapogenins exhibited a similar permeability to that of saponins, although the digestion enhanced the permeability of sapogenins from quinoa (1.14 ± 0.47 × 10-5 cm/s) but not from fenugreek (2.33 ± 0.99 × 10-6 cm/s). An overall positive impact of coexisting lipids on the permeability was evidenced. PAMPA is shown as a useful, rapid, and easy tool for assessing the permeability of bioactive compounds from complex matrices, with the previous gastrointestinal process being a relevant step.

Fenugreek Counters the Effects of High Fat Diet on Gut Microbiota in Mice: Links to Metabolic Benefit.

Fenugreek (Trigonella foenum-graecum) is an annual herbaceous plant and a staple of traditional health remedies for metabolic conditions including high cholesterol and diabetes. While the mechanisms of the beneficial actions of fenugreek remain unknown, a role for intestinal microbiota in metabolic homeostasis is likely. To determine if fenugreek utilizes intestinal bacteria to offset the adverse effects of high fat diets, C57BL/6J mice were fed control/low fat (CD) or high fat (HFD) diets each supplemented with or without 2% (w/w) fenugreek for 16 weeks. The effects of fenugreek and HFD on gut microbiota were comprehensively mapped and then statistically assessed in relation to effects on metrics of body weight, hyperlipidemia, and glucose tolerance. 16S metagenomic analyses revealed robust and significant effects of fenugreek on gut microbiota, with alterations in both alpha and beta diversity as well as taxonomic redistribution under both CD and HFD conditions. As previously reported, fenugreek attenuated HFD-induced hyperlipidemia and stabilized glucose tolerance without affecting body weight. Finally, fenugreek specifically reversed the dysbiotic effects of HFD on numerous taxa in a manner tightly correlated with overall metabolic function. Collectively, these data reinforce the essential link between gut microbiota and metabolic syndrome and suggest that the preservation of healthy populations of gut microbiota participates in the beneficial properties of fenugreek in the context of modern Western-style diets.

The Molecular Mechanisms of Regulating Oxidative Stress-Induced Ferroptosis and Therapeutic Strategy in Tumors.

Ferroptosis is an atypical form of regulated cell death, which is different from apoptosis, necrosis, pyroptosis, and autophagy. Ferroptosis is characterized by iron-dependent oxidative destruction of cellular membranes following the antioxidant system's failure. The sensitivity of ferroptosis is tightly regulated by a series of biological processes, the metabolism of iron, amino acids, and polyunsaturated fatty acids, and the interaction of glutathione (GSH), NADPH, coenzyme Q10 (CoQ10), and phospholipids. Elevated oxidative stress (ROS) level is a hallmark of cancer, and ferroptosis serves as a link between nutrition metabolism and redox biology. Targeting ferroptosis may be an effective and selective way for cancer therapy. The underlying molecular mechanism of ferroptosis occurrence is still not enough. This review will briefly summarize the process of ferroptosis and introduce critical molecules in the ferroptotic cascade. Furthermore, we reviewed the occurrence and regulation of reduction-oxidation (redox) for ferroptosis in cancer metabolism. The role of the tumor suppressor and the epigenetic regulator in tumor cell ferroptosis will also be described. Finally, old drugs that can be repurposed to induce ferroptosis will be characterized, aiming for drug repurposing and novel drug combinations for cancer therapy more efficiently and economically.

In Vitro Colonic Fermentation of Saponin-Rich Extracts from Quinoa, Lentil, and Fenugreek. Effect on Sapogenins Yield and Human Gut Microbiota.

In vitro colonic fermentation of saponin-rich extracts from quinoa, lentil, and fenugreek was performed. Production of sapogenins by human fecal microbiota and the impact of extracts on representative intestinal bacterial groups were evaluated. The main sapogenins were found after fermentation (soyasapogenol B for lentil; oleanolic acid, hederagenin, phytolaccagenic acid, and serjanic acid for quinoa; and sarsasapogenin, diosgenin, and neotigogenin acetate for fenugreek). Interindividual differences were observed, but the highest production of sapogenins corresponded to quinoa (90 µg/mL) and fenugreek (70 µg/mL) extracts, being minor for lentil (4 µg/mL). Lentil and quinoa extracts showed a general antimicrobial effect, mainly on lactic acid bacteria and Lactobacillus spp. Significant increases of Bifidobacterium spp. and Lactobacillus spp. were observed for fenugreek in one volunteer. Thus, the transformation of saponin-rich extracts of quinoa, lentil, and fenugreek to sapogenins by human gut microbiota is demonstrated, exhibiting a modulatory effect on the growth of selected intestinal bacteria.

CO2 treatment improves the hypocholesterolemic and antioxidant properties of fenugreek seeds.

A high level of serum cholesterol is a major cause of atherosclerosis. Fenugreek is a well-known hypocholesterolaemic agent with amazing phytochemical composition. Due to its impact on plant metabolism, CO2 enrichment was tested as a strategy to support functional values in fenugreek seeds. Phytochemical composition and biological activities of three fenugreek cultivars (G2, G6 and G30) grown under ambient (aCO2, 400 µmol mol-1) and elevated CO2 (eCO2, 620 µmol mol-1) were assessed. Applying eCO2 improved physical parameters of fenugreek seeds, and enhanced their biological activities. A significant increase in hypocholesterolaemic potential, as indicated by inhibition of cholesterol micellar solubility and pancreatic lipase activity, was recorded. In addition, antioxidant, anti-lipid peroxidation and antibacterial activities were improved. These enhanced biological activities were accompanied by improved seed chemical composition at the primary and secondary metabolic levels. Therefore, eCO2 treatment represents an efficient strategy to increase the hypocholesterolaemic, antioxidant and antibacterial activities of fenugreek seeds.

Effect of planting time and vermicompost on the proteomic pattern of fenugreek (Trigonella foenum-graecum)

The fenugreek is one of the most important medicinal plants belongs to Fabaceae, originated in West Asia, Iran and Mediterranean regions. This research included a qualitative study of fenugreek proteins using SDS-PAGE electrophoresis on polyacrylamide gel and the separation of protein bands of fenugreek leaves in different treatments of vermicompost fertilizer and cultivating dates. Results showed that a band (about 80 kDa) on the first planting date (May 31) is observed in all samples except for sample a1 (10 t/ha vermicompost on May 31). Another significant difference was the band contained in the third planting date (31 September) and in the molecular weight of about 15 kDa, which was not seen in other dates. This difference can be due to the synthesis of this protein with the mentioned weight under the conditions of reducing the temperature in the early fall. It also showed more differences in two-dimensional electrophoresis, for example, in 14 kDa and PI in the range of 4.5-4.7 in treatment without fertilizer, no protein expression was observed, which was consistent with the results of the SDS-PAGE test.

Biosynthesis of iron nanoparticles using Trigonella foenum-graecum seed extract for photocatalytic methyl orange dye degradation and antibacterial applications.

Trigonella foenum-graecum is the source of various biological and chemical constituents with a wide area of applications, especially in the treatment/prevention of diabetes and other chronic diseases such as cancer. Multiple biological and organic moieties in the aqueous or the organic phase of Trigonella foenum-graecum carry soft reduction properties to reduce the metal cations to nanoparticles. In this investigation, the Trigonella foenum-graecum was found in the seed extract for the first time in an aqueous medium. We successfully synthesized zero-valent iron nanoparticles (Fe0) (ZV-Fe NPs) and stabilized these nanoparticles in an aqueous medium. The stabilization mechanism of Fe NPs by Trigonella foenum-graecum in an aqueous extract was investigated. Further, Fe NPs were characterized by UV-visible spectrometry, X-ray diffraction (XRD), thermogravimetric analysis - derivative thermo-gravimetric (TGA/DTG), magnetization, Fourier transform infrared (FTIR) spectroscopy and transmission electron microscopy (TEM) images. The size of the nanoparticles, calculated using the Debye-Scherer equation and TEM, was found to be approximately 11 nm with the highest particle distribution number. Fe NPs are very effective for methyl orange dye degradation under UV light following pseudo first-order kinetics, and the rate constant kapp was found to be 0.025 min-1. Furthermore, Fe NPs were applied to check the antibacterial activities with microorganisms such as gram-negative E. coli and gram-positive S. aureus. The Minimum Inhibitory Concentration (MIC) of Fe NPs for E. coli and S. aureus was calculated as 32 µg/mL and 64 µg/mL, respectively.