Differential responses of two fenugreek (Trigonella foenum-graecum L.) landraces pretreated with melatonin to prolonged drought stress and subsequent recovery.

BACKGROUND: Drought impairs growth, disturbs photosynthesis, and induces senescence in plants, which results in crop productivity reduction and ultimately jeopardizes human food security. The objective of this study was to determine major parameters associated with drought tolerance and recovery ability of fenugreek (Trigonella foenum-graecum L.), by examining differential biochemical and phenological responses and underlying enzyme activities as well as melatonin roles during drought stress and re-watering for two contrasting landraces. Moreover, the relative expression of three key genes involved in the biosynthesis pathway of diosgenin, including SQS, CAS, and BG, was investigated. RESULTS: Depending on the conditions, drought stress enhanced the activity of antioxidant enzymes and the osmoregulating compounds, non-enzymatic antioxidants, hydrogen peroxide content, and lipid peroxidation levels in most cases. Severe drought stress accelerated flowering time in Shushtar landrace (SHR) but had no significant effects on Varamin (VR). Pretreatment with melatonin delayed flowering time in SHR and caused high drought resistance in this landrace. Furthermore, melatonin significantly enhanced drought adaptability in VR by improving plant recovery ability. DISCUSSION: Based on our results plants' responses to drought stress and melatonin pretreatment were completely landrace-specific. Drought stress caused an increase in the relative expression of CAS gene and ultimately the accumulation of steroidal saponins in SHR. Melatonin compensated for the decrease in biomass production due to drought stress and finally increased steroidal saponins performance in SHR. Our study showed that melatonin can improve drought stress and recovery in fenugreek, but different factors such as genotype, melatonin concentration, and plant age should be considered.

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.

Application of fenugreek in ruminant feed: implications for methane emissions and productivity.

Background: Human demand for meat and dairy products will increase as a result of economic development and population growth, and the farming of ruminants, such as cattle and sheep, will also increase. Methane (CH4) emission from the enteric fermentation of ruminant livestock is a major source of greenhouse gas emissions and a significant contributor to global warming. Meanwhile, growth performance is often limited and animals are more vulnerable to diseases in high-density, intensive farming, greatly reducing livestock productivity, so developing ways to reduce CH4 emissions and improve ruminant productivity has become a research hotspot. Studies have reported that fenugreek (Trigonella foenum-graecum L.) as feed additives have the potential to reduce ruminant methane and improve the productivity. However, systematic reviews of such studies are lacking. Methodology: In this review, databases of Google Scholar, Web of Science, PubMed, Scopus and Science Direct were used for the literature search. The initial keywords search was fenugreek or Trigonella foenum-graecum L. For more focused search, we added terms such as methane, rumen fermentation, growth, milk production and antioxidants. All were done for ruminants. The literature that conforms to the theme of this article is selected, summarized, and finally completed this article. Results: By regulating the rumen microbiome (suppressing protozoans, methanogenic bacteria, and fungi), fenugreek can lower CH4 emissions according to many in vitro anaerobic fermentation experiments. Fenugreek secondary metabolites (saponins and tannins) are responsible for this impact, but it is still unclear exactly how they work. Therefore, more long-term in vivo experiments are needed to verify its efficacy. Fenugreek is also rich in alkaloids, amino acids, flavonoids, saponins and phenolic acids. These compounds have been shown to have beneficial effects on ruminant growth, lactation, and total antioxidant capacity. Therefore, fenugreek has a great opportunity to develop into a new green feed additive. Conclusions: This review provides a summary of the effect of fenugreek and its bioactive compounds on rumen fermentation, CH4 emissions and production performance by ruminants. In addition, based on the available data, the possible biochemical pathway of fenugreek to reduce CH4 emissions in ruminants was described. Overall, the livestock feed industry has the opportunity to develop natural, environmentally-friendly feed additives based on fenugreek.

The amelioration of salt stress-induced damage in fenugreek through the application of cold plasma and melatonin.

Nowadays, it is increasingly crucial to combine innovative approaches with established methods to enhance plant tolerance and maximize the production of beneficial compounds. With this aim in view, a study was carried out to investigate how different melatonin concentrations (0, 30, and 60 ppm), cold plasma treatment (at 3000 and 4000 V), and varying exposure durations (0, 1, 2, and 4 min) affect the physiological and biochemical attributes of fenugreek plants, as well as the levels of diosgenin under salinity stress. This study revealed that the application of 3000 V cold plasma for 2 min with 60 ppm melatonin by establishing cellular redox homeostasis in salinity-treated fenugreek plants, effectively prevented the destruction of pigments and reduced the electrolyte leakage index of malondialdehyde content. The utilization of these two elicitors has the potential to trigger multiple pathways, including the enzymatic and non-enzymatic antioxidants biosynthesis, and abscisic acid-dependent pathways. This activation results in an enhanced production of abscisic acid, auxin, and endogenous melatonin, along with the regulation of signal transduction pathways. Surprisingly, applying these two treatments increased the expression of SQS, CAS, SSR, BGL, SEP, SMT, and diosgenin content by 13, 22.5, 21.6, 19, 15.4, 12, and 6 times respectively. The findings highlight the intricate interplay between these treatments and the positive impact of their combined application, opening up avenues for further research and practical applications in improving plant tolerance to environmental stresses.

Evaluating the hidden potential of deashed biochar in mitigating salinity stress for cultivation of fenugreek.

Soil salinity, the second most prominent cause of land degradation after soil erosion, has posed a persistent challenge to agriculture. Currently, approximately 1 billion hectares of Earth's land surface, equivalent to 7%, are affected by salinity. While biochar has proven effective in mitigating salinity stress, the specific role of deashed biochar in salinity mitigation has not been thoroughly explored. Therefore, this study was conducted to investigate the impact of four levels of deashed biochar (0%, 0.4%, 0.8%, and 1.2%) on the growth and physiological attributes of Fenugreek under both non-saline conditions (2.54 dS/m EC) and salinity stress conditions (5.46 dS/m EC). The results revealed a notable enhancement in various parameters under salinity stress. Compared to the control, the application of 1.20% deashed biochar led to a significant increase in shoot fresh weight (30.82%), root fresh weight (13.06%), shoot dry weight (17.43%), root dry weight (33.44%), shoot length (23.09%), and root length (52.39%) under salinity stress. Furthermore, improvements in internal CO2 concentration (9.91%), stomatal conductance (15.49%), photosynthetic rate (25.50%), and transpiration rate (10.46%) were observed, validating the efficacy of 1.20% deashed biochar in alleviating salinity stress. The study also demonstrated a significant decrease in the activities of oxidative stress markers such as peroxidase (POD), superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), electrolyte leakage, and malondialdehyde (MDA). Simultaneously, there was an increase in the concentrations of essential nutrients, namely nitrogen (N), phosphorus (P), and potassium (K), in both shoot and root tissues. These findings collectively suggest that deashed biochar, particularly at a concentration of 1.20%, is recommended for achieving enhanced crop production under conditions of 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.

Exogenous abscisic acid fine-tunes heavy metal accumulation and plant's antioxidant defence mechanism to optimize crop performance and secondary metabolite production in Trigonella foenum-graecum L. under nickel stress.

Nickel (Ni) contamination of farming soil has become currently a recurring global menace to agriculture crop productivity. The purpose of the present study was to investigate the putative contributions of abscisic acid (ABA) to extemporize Ni tolerance in Trigonella foenum-graecum L. (fenugreek) plants. The outcomes of this study exposed that exogenous supplementation of ABA at 10, 20, 40 and 80 µM considerably enhanced the growth and physiological attributes of fenugreek under 80 mg Ni kg-1 soil, however, 40 µM of ABA exhibited the best results under normal and Ni-stressed conditions. ABA-mediated Ni tolerance was marked by reductions in Ni accumulation and consequent lowering of reactive oxygen species (ROS) like hydrogen peroxide and superoxide radicals. Contrarily, NO (nitric oxide) level increased in response to ABA application under Ni stress conditions, accompanied by promoted antioxidant activities through improved levels of secondary metabolites, proline, and perked-up ROS-detoxification enzymes activities. Exogenous ABA at 40 µM concentration applied to Ni-exposed plants (80 mg Ni kg-1 soil) improved the total content of alkaloids, phenolics, flavonoids and tannins by 14.3%, 10.2%, 15.4% and 7.0%, respectively, over Ni-stressed plants alone. Additionally, seed trigonelline content imparting several pharmacological actions to the fenugreek plant exhibited a remarkable escalation upto 3.6 and 2.6 mg g-1 DW under '40 µM ABA' and '40 µM ABA + 80 mg Ni kg-1 soil' treatments, respectively. The findings of the study suggest that ABA plays a key role in enhancing the overall performance of the fenugreek crop under excessive Ni stress.

Exogenous 24-epibrassinolide ameliorates tolerance to high-temperature by adjusting the biosynthesis of pigments, enzymatic, non-enzymatic antioxidants, and diosgenin content in fenugreek.

High-temperature stress is widely considered a main plant-growth-limiting factor. The positive effects of 24-epibrassinolide (EBR) as analogs of brassinosteroids (BRs) in modulating abiotic stresses have led this hormone to be referred to as a growth regulator in plants. The current study highlights the influence of EBR on enhancing tolerance to high-temperature and altering the diosgenin content in fenugreek. Different amounts of EBR (4, 8, and 16 µM), harvesting times (6, and 24 h), as well as temperature regimes (23 °C, and 42 °C) were, used as treatments. EBR application under normal temperature and high-temperature stress resulted in decreased malondialdehyde content and electrolyte leakage percentage, while the activity of antioxidant enzymes improved significantly. Exogenous EBR application possibly contributes to activating the nitric oxide, H2O2, and ABA-dependent pathways, enhancing the biosynthesis of abscisic acid and auxin, and regulating the signal transduction pathways, which raises fenugreek tolerance to high-temperature. The SQS (eightfold), SEP (2.8-fold), CAS (11-fold), SMT (17-fold), and SQS (sixfold) expression, considerably increased following EBR application (8 µM) compared to the control. Compared to the control, when the short-term (6 h) high-temperature stress was accompanied by EBR (8 µM), a sixfold increase in diosgenin content was achieved. Our findings highlight the potential role of exogenous 24-epibrassinolide in mitigating the high-temperature stress in fenugreek by stimulating the biosynthesis processes of enzymatic and non-enzymatic antioxidants, chlorophylls, and diosgenin. In conclusion, the current results could be of utmost importance in breeding or biotechnology-based programs of fenugreek and also in the researches related to the engineering of the biosynthesis pathway of diosgenin in this valuable plant.

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.

Soil-applied Nickel Generates Differential Responses in Growth, Physiology and Oxidative Metabolism of Fenugreek (Trigonella foenum-graecum L.) Varieties.

This work aims to evaluate the potential of nickel (Ni), an essential micronutrient, as an oxidative stress inducer along with associated morphological and biochemical responses in different varieties of fenugreek (Trigonella foenum-graecum L.), a chief economically cultivated crop of India. Varietal differences in crop performance upon exposure to 0, 20, 40, 60 and 80 mg Ni kg- 1 soil reflects that Ni applied at 20 mg Ni kg- 1 soil offers growth-promoting effects, improved photosynthesis attributes, carbonic and nitrate reductase activities more profound in PEB followed by AFg2, AFg1 and UM185 variety. This study observed a dose-dependent reduction in all the above parameters. Maximum toxic effects were noticed at 80 mg kg- 1 Ni, manifested in the form of enhanced H2O2 and MDA contents, which were efficiently counteracted by augmentation in proline content, SOD, POX, CAT and APX activities in PEB over other varieties, suggesting that the Ni tolerance in fenugreek varieties can be organized as PEB > AFg2 > AFg1 > UM185.

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.

Metabolic engineering of the diosgenin biosynthesis pathway in Trigonella foenum-graceum hairy root cultures.

Diosgenin as a triterpene with numbers of pharmaceutical applications has been identified in Trigonella foenum-graceum. In this survey, in order to scale up the amount of diosgenin in Fenugreek as a promising alternative of yam, ∆24-reductase as a rate limiting enzyme in diosgenin biosynthesis pathway has been overexpressed by utilizing pBI121 expression plasmid in hairy roots culture platform. The recombinant binary vector pBI121-∆24-reductase was transformed into R. rhizogenes strain ATCC 15834 to induce transgenic hairy roots in "Hamedan" as a low-diosgenin production genotype. In the transgenic hairy roots, the ∆24-reductase expression level was significantly 8.15 times overexpressed comparing to the non-transgenic hairy roots, Nonetheless the Sterol-methyltransferase, as a competitive enzyme, was 6 times downregulated. Furthermore, the expression rate of Squalene synthase, Cycloartenol synthase, C26-Hydroxylase were also increased 1.5, 1.7, 2.9 times higher than those of the non-transgenic hairy roots, respectively. The diosgenin content in the transgenic hairy root was raised 3 times up comparing to the non-transgenic hairy roots, besides it was scaled up 25-fold comparing to the diosgenin amount in "Hamedan" Leaf. As a result, the first metabolic engineering on this pathway was clearly revealed the impact of ∆24 -reductase gene in diosgenin content enhancement.

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.

Priming with EDTA, IAA and Fe alleviates Pb toxicity in Trigonella Foneum graecum L. growth: Phytochemicals and secondary metabolites.

This study evaluated the effects of the exogenous application of ethylenediaminetetraacetic acid (EDTA), indole-3-acetic acid (IAA) and iron sulfate (FeSO4) upon the phytochemical mechanisms of fenugreek grown under Pb-excess (2000 mg L-1 PbCl2). The results showed that chemical additives of EDTA and IAA as well as FeSO4 improved fenugreek germination parameters. The radicle length and the amylase activity were significantly improved under IAA treatment compared to EDTA and FeSO4. Exogenous FeSO4 was more effective to improving growth parameters. Moreover, the decrease in hydrogen peroxide (H2O2) and malondialdehyde (MDA) levels was noted under all chemical additives especially under IAA application. In addition, it was more effective than EDTA and Fe in increasing catalase, glutathione (GSH), ascorbate peroxidase (APX), flavonoids and phenols while the increment superoxide dismutase (SOD) production was more pronounced under EDTA addition to Pb than other chelators. HPLC analysis revealed that the gallic was the major phenol produced under all chelators addition especially with IAA. In addition, the syringic acid was only produced with exogenous IAA while the quercetin was only detected under EDTA addition. Our results exhibited a higher IAA efficiency than EDTA and FeSO4 in mitigating Pb stress in fenugreek through up-regulated mechanisms of the antioxidant system for reducing reactive oxygen species (ROS) activities and enhancing special phenols.

[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.

Hydrogen sulphide alleviates cadmium stress in Trigonella foenum-graecum by modulating antioxidant enzymes and polyamine content.

Cadmium (Cd) toxicity reduces growth and yield of crops grown in metal-polluted sites. Research was conducted to estimate the potential of hydrogen sulphide (H2 S) to mitigate toxicity caused by Cd in fenugreek seedlings (Trigonella foenum-graecum L.). Different concentrations of CdCl2 (Cd1-1 mM, Cd2-1.5 mM, Cd3-2mM) and H2 S (HS1-100 µM, HS2-150 µM, HS3-200 µM) were assessed. Seeds of fenugreek were primed with sodium hydrosulphide (NaHS), as H2 S donor. Seedlings growing in Cd-spiked media treated with H2 S were harvested after 2 weeks. Cd stress affected growth of fenugreek seedlings. Cd toxicity decreased leaf relative water content (LRWC), intercellular CO2 concentration, net photosynthesis, stomatal conductance and transpiration. However, application of H2 S significantly improved seedling morphological attributes by increasing the activity of antioxidant enzymes, i.e. APX, CAT and SOD, in Cd-contaminated soil. H2 S treatment also regulated phenolic and flavonoid content. H2 S-induced biosynthesis of spermidine (Spd) and putrescine (Put) could account for the enhancement of growth and physiological performance of fenugreek seedlings under Cd stress. H2 S treatment also reduced H2 O2 production (38%) and electrolyte leakage (EL, 51%) in seedlings grown in different concentrations of Cd. It is recommended to evaluate the efficacy of H2 S in alleviating Cd toxicity in other crop plants.

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.

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.

Preparation and characterization of fenugreek leaf flavonoids and their protective effects against oxidative damage to hepatocytes / 中国中药杂志

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.