Search Results (44)

Drought stress significantly hinders the cultivation of medicinal plants such as licorice (Glycyrrhiza uralensis), valued for its bioactive compounds, glycyrrhizin, and liquiritin. This study aims to investigate how co-inoculation with arbuscular mycorrhizal fungus Rhizophagus irregularis and Trichoderma harzianum can enhance licorice drought tolerance and secondary metabolite production, providing insights for sustainable agriculture in arid regions. The results demonstrate that inoculation with R. irregularis significantly improved biomass, drought stress tolerance, and increased glycyrrhizin and liquiritin concentrations by 29.9% and 3.3-fold, respectively, particularly under drought conditions. Co-inoculation with T. harzianum further boosted glycyrrhizin yield by 93.7%, indicating a synergistic relationship between the two microbes. The expression of key biosynthetic genes, including squalene synthase (SQS1) for glycyrrhizin and chalcone synthase (CHS) for liquiritin, was significantly upregulated, enhancing water use efficiency and the biosynthesis of secondary metabolites. Nutrient analysis showed improved phosphorus uptake, alongside reduced root carbon and nitrogen concentrations, leading to greater nutrient utilization efficiency. These findings suggest that co-inoculating R. irregularis and T. harzianum is a promising approach to improving licorice growth and medicinal quality under drought stress, with broad applications for sustainable crop management. Full article

Background: Fine dust exposure worsens oily skin by disrupting lipid metabolism and triggering oxidative inflammation. Scutellaria baicalensis extract-induced exosomes (SBEIEs) have shown anti-inflammatory effects by suppressing reactive oxygen species (ROS) and lipid-regulating properties, making them potential therapeutic agents. Methods: Exosomes from fibroblasts treated with SBEIEs and PM10 were tested on macrophages, adipose-derived stem cells (ASCs), and T lymphocytes. ELISA, flow cytometry, and PCR measured cytokines and gene expression. A 10-day clinical trial evaluated skin hydration, oiliness, and inflammation. Results: SBEIEs increased IRF3 (1.6 times) and suppressed PPARγ in ASCs while enhancing lipolysis markers. Sebaceous gland activity (squalene synthase) decreased by 10%. Macrophages showed increased IRF3, IFN-β, and IL-10 (2.1 times). T cells secreted IL-4 and IL-22 (2–2.33 times). Clinically, SBEIEs improved hydration (21%), reduced oiliness (1.6 times), and decreased inflammation (2.2 times). Conclusions: SBEIEs effectively regulate lipid metabolism, cytokines, and immune responses, showing promise to treat oily and inflamed skin caused by fine dust exposure. Further studies are needed for clinical applications. Full article

This publication reports the controlled cultivation of Zanthoxylum chiloperone var. angustifolium Engl. (Rutaceae) in several growth substrates under controlled greenhouse conditions. This plant is well-known for its anti-Chagas (trypanocidal) activity, related to the presence of several β-carboline alkaloids. The metabolomic study of Z. chiloperone seedlings over two years of growth (2018–2020) was performed using comprehensive two-dimensional gas chromatography time-of-flight mass spectrometry (GC × GC-TOFMS). The canthin-6-one alkaloids, canthin-6-one and 5-methoxy-canthin-6-one, were putatively identified in Z. chiloperone extracts. Finally, in vitro and in silico studies of trypanocidal activity were performed, suggesting that canthin-6-one alkaloids could interact with the main pharmacological targets against Trypanosoma cruzi, cruzain protease, dihydroorotate dehydrogenase, lanosterol 14-alpha-demethylase, farnesyl diphosphate, and squalene synthases. Full article

Ergosterol is a component of fungal cell membranes that has physiological functions and applications in drugs, such as anti-inflammatory, anti-tumor, anti-fungal, and other immunosuppressive activities. The fungus Athelia termitophila, also known as the termite ball fungus, primarily contains secondary metabolites (like active ingredients) that are similar to ergosterol. To enhance the synthesis of ergosterol and mycelial biomass in termite ball fungus, methyl jasmonate and salicylic acid were used to stimulate the biosynthesis of ergosterol compounds during the growth of TMB mycelium and relative quantitative levels of gene transcripts. The conditions of the inducers were optimized. Under 80 µmol/L MJ incubation conditions, the content of ergosterol compounds in TMB was increased by 2.23-fold compared with the wild-type strain. RT-qPCR results at the transcriptional level of ergosterol synthesis pathway genes showed that MJ significantly induced the expression of HMGR (3-Hydroxy-3-Methylglutaryl-Coa Reductase), HMGS (3-Hydroxy-3-Methylglutaryl-Coa Synthase), SE (Squalene Epoxidase), and FPS (Farnesyl Diphosphate Synthase) genes in the ergosterol synthesis pathway. For expression levels at different induction days, we collected 7/10 d and 4/6/8 d samples with similar expression patterns, as well as SS (Squalene Synthase)/FPS (Farnesyl Diphosphate Synthase), SE (Squalene Epoxidase)/MVD (Mevalonate Diphosphate Decarboxylase), and HMGS (3-Hydroxy-3-Methylglutaryl-Coa Synthase)/HMGR (3-Hydroxy-3-Methylglutaryl-Coa Reductase) genes with similar expression patterns, which resulted in gene transcription data during ergosterol content synthesis. The current study provides an effective method to increase the ergosterol contents in termite ball fungus and a new idea for the mechanism of MJ-induced ergosterol compound biosynthesis. Full article

This study presents the first comprehensive genomic analysis of Cryptoporus qinlingensis, a classical folk medicine and newly identified macrofungus from the Qinling Mountains. Utilizing advanced sequencing technologies, including PacBio HiFi and Hi-C, we achieved a high-quality chromosome-level genome assembly. The genome, sized at 39.1 Mb, exhibits a heterozygosity of 0.21% and contains 21.2% repetitive sequences. Phylogenetic analysis revealed a recent divergence of C. qinlingensis from Dichomitus squalens approximately 212.26 million years ago (MYA), highlighting the rapid diversification within the Polyporaceae family. Comparative genomic studies indicate significant gene family contraction in C. qinlingensis, suggesting evolutionary adaptations. The identification of a tetrapolar mating system, along with the analysis of CAZymes and P450 genes, underscores the genomic complexity and ecological adaptability of this species. Furthermore, the discovery of 30 biosynthetic gene clusters (BGCs) related to secondary metabolites, including polyketide synthase (PKS), non-ribosomal peptide synthetase (NRPS), and terpene synthesis enzymes, opens new avenues for exploring bioactive compounds with potential medicinal applications. This research not only enriches our understanding of the Cryptoporus genus but also provides a valuable foundation for future studies aiming to harness the therapeutic potential of C. qinlingensis and to further explore its ecological and evolutionary significance. Full article

Background/Objectives: Developing antifungal drugs with lower potential for interactions with food may help to optimize treatment and reduce the risk of antimicrobial resistance. Chemometrics uses statistical and mathematical methods to analyze multivariate chemical data, enabling the identification of key correlations and simplifying data interpretation. We used the partial least squares (PLS) approach to explore the correlations between various characteristics of oral antifungal drugs (including antifungal antibiotics) and dietary interventions, aiming to identify patterns that could inform the optimization of antifungal therapy. Methods: We analyzed 15 oral antifungal drugs, including azoles (8), antifungal antibiotics (4), antifungal antimetabolites (1), squalene epoxidase inhibitors (1), and glucan synthase inhibitors (1). The input dataset comprised information from published clinical trials, chemical records, and calculations. We constructed PLS models with changes in the pharmacokinetic parameters (∆AUC, area under the curve; ∆C_max, maximum drug concentration; and ∆T_max, time to reach maximum drug concentration) after dietary intervention as the response parameters and eight groups of molecular descriptors (M1–M8) as the predictor parameters. We performed separate analyses for the different nutritional interventions. Results: In the final PLS model with food as an intervention, we effectively reduced the dimensionality of the dataset while retaining a substantial percentage of the original information (variance), as significant components explained 69.8% and 17.5% of the predictor and response parameter variances, respectively. The PLS model was significant because its components met the cross-validation criteria. We obtained six significant positive and negative correlations between the descriptors related to atoms and the postprandial ∆T_max. Conclusions: The PLS method is valuable for investigating interactions between antifungal drugs (including antifungal antibiotics) and food. The correlations obtained can be used in drug modeling to predict interactions with dietary interventions based on the antifungal drug’s chemical structure. Incorporating chemometric techniques into the early drug development stages could facilitate the design of antifungal antibiotics and other antifungal agents with optimized absorption in the presence of dietary components. Full article

Squalene synthase (SQS) is the most direct key enzyme regulating squalene synthesis. To better understand the regulatory mechanisms of squalene biosynthesis, a 1423-bp long promoter region of the CoSQS gene was isolated from Camellia oleifera. Plant CARE and PLACE analysis affirmed the existence of the core promoter elements such as TATA and CAAT boxes and transcription factor binding sites like W-box and MYB in the isolated sequence. Exogenous factors regulating the CoSQS promoter were obtained by using Yeast one-hybrid screening, and the key transcription factor CoWRKY15 was found. AOS (Antibody Optimization System) analysis showed that CoWRKY15 had the highest interactions with a confidence level of 0.9026. Bioinformatics analysis showed that CoWRKY15 belonged to class 2 of the WRKY gene family. The results of subcellular localization showed that CoWRKY15 functioned in the nucleus. The results of CoWRKY15 promoter analysis showed that 8 out of 14 cis-elements with annotatable functions were related to the light response. The region of the CoSQS promoter that interacts with CoWRKY15 is −186 bp~−536 bp. The histochemical assay and squalene content suggested that the CoSQS promoter could drive the expression of GUS gene and specific promotion of CoSQS expression. It was found that CoWRKY15 could act on the −186 bp~−536 bp CoSQS promoter to regulate the expression of CoSQS and the content of squalene in C. oleifera seed kernels. Full article

Paris polyphylla Smith var. Chinensis (Franch.) Hara is a medicinal plant that belongs to the Liliaceae family. Its main components are parissaponins, which have excellent medicinal effects such as anti-inflammatory, anti-tumor, etc. Improving the quality of parissaponins through artificial directional regulation has emerged as a practice to meet medical demand and is a new research hotspot. In this paper, P. polyphylla plants were treated with UVA, UVB, and UVC, and the contents of PolyPhyllin VI (PPVI) and PolyPhyllin VII (PPVII), saponin synthase (squalene synthase, SS; cycloartenol synthase, CAS; cytochrome P450, CYP450; and glycosyl transferases, GT) activity, MDA, and the photosynthetic pigment indexes were measured and analyzed. The results showed that PPVII content increased by 32.43% with UVC treatment after 4 h (3.43 mg/g), but the PPVI and PPVII contents in the other groups decreased compared with CK (control group) and they did not return to the original level after 4 h. SS, CAS, CYP450, and GT synthases were activated in varying degrees via UV treatment and increased, respectively, by 22.93%, 10.83%, 20.15%, and 25.98%. Among them, GT, as the last of the synthetases, had a shorter response time to UVB (30 min) and UVC (15 min); the difference was sensible compared with CK. Moreover, UV had a stressing effect and promoted the rapid accumulation of MDA content (increased 17.66%, 34.53%, and 9.65%) and carotenoid (increased 7.58, 5.60, and 7.76 times) within 4 h compared to CK. UVB and UVC radiation visibly improved chlorophyll a content (42.56% and 35.45%), but UVA did not, and the change in chlorophyll b content showed no overt statistical difference. In addition, PPVI and PPVII were negatively correlated with SS, CAS, carotenoids, and MDA (p < 0.05) and positively correlated with CYP450, GT, and chlorophyll a (p < 0.05). This study provides a theoretical basis for using UV light to regulate secondary metabolism in P. polyphylla, which is of great value for production management. Full article

The accumulation of ginsenosides (triterpenic saponins) was determined in Panax quinquefolium hairy root cultures subjected to an elicitation process using carvacrol at 5, 10, 25, 50, 100, 250, and 500 μM concentrations during 24 and 72 h exposure. This study was the first one in which carvacrol was applied as an elicitor. The content of eight ginsenosides, Rb1, Rb2, Rb3, Rc, Rd, Rg1, Rg2, and Re, was determined using HPLC analysis. Moreover, the quantitative RT-PCR method was applied to assess the relative expression level of farnesyl diphosphate synthase, squalene synthase, and dammarenediol synthase genes in the studied cultures. The addition of carvacrol (100 μM) was an effective approach to increase the production of ginsenosides. The highest content and productivity of all detected saponins were, respectively, 20.01 mg∙g⁻¹ d.w. and 5.74 mg∙L⁻¹∙day⁻¹ after 72 h elicitation. The production profile of individual metabolites in P. quinquefolium cultures changed under the influence of carvacrol. The biosynthesis of most examined protopanaxadiol derivatives was reduced under carvacrol treatment. In contrast, the levels of ginsenosides belonging to the Rg group increased. The strongest effect of carvacrol was noticed for Re metabolites, achieving a 7.72-fold increase in comparison to the control. Saponin Rg2, not detected in untreated samples, was accumulated after carvacrol stimulation, reaching its maximum concentration after 72 h exposure to 10 μM elicitor. Full article

The genome sequence of Leishmania has given rise to diverse novel drug targets, and their identification remains the first step in drug discovery. This study aims to identify the possible anti-leishmanicidal activity target(s) of N¹, N⁴-[dibenzylbutane-4′,4″-(dioxymethylenebenzene)]-1,4-diamine from a plethora of pathways in kinetoplastids. The compound was docked using AutoDockTools-1.5.6 against eight co-crystallized proteins selected from the protein data bank, each representing important biosynthetic pathways. The evaluation of the best conformational protein–ligand poses showed that the N, N′-substituted diamine binds more efficiently to glyceraldehyde-3-phosphate dehydrogenase (G3PDH) (E = −8.97 Kcal/mol and Ki = 0.267 µM; Ki co-crystallized ligand = 19.39 µM), which is responsible for the conversion of glyceraldehyde-3-phosphate to 1,3-bisphosphoglycerate and pteridine reductase I (PTR1) (E = −8.75 Kcal/mol and Ki = 0.387 µM; Ki co-crystallized ligand = 60.56 µM), which reduces both pterins and folates to tetrahydrobiopterin and tetrahydrofolate, respectively. Moderate binding activity by the ligand was observed for the protein kinases (CDKs) (E = −8.37 Kcal/mol and Ki = 0.729 µM; Ki co-crystallized ligand = 26.80 µM) and trypanothione reductase (TR) (E = −8.57 Kcal/mol and Ki = 0.525 µM; Ki co-crystallized ligand = 174.68 µM) of the trypanothione biosynthetic pathway. With E > −7.35 Kcal/mol and Ki > 4.10 µM, the ligand appears to have no significant inhibition of the squalene synthase (SQS), lactoyl glutathione lyase (LGL) or the pteridine synthase (TS) of the sterol, glyoxalase and trypanothione biosynthetic pathways. The efficient inhibition of G3PDH and PTR1 targets in Leishmania by N, N′-substituted diamine molecule provides more insights into understanding the mechanism of leishmanicidal activity. Full article

Centella asiatica is a medicinal plant with a rich tradition of use for its therapeutic properties. Among its bioactive compounds are centellosides, a group of triterpenoid secondary metabolites whose potent pharmacological activities have attracted significant attention. Metabolic engineering has emerged as a powerful biotechnological tool to enhance the production of target compounds. In this study, we explored the effects of overexpressing the squalene synthase (SQS) gene and transcription factor TSAR2 on various aspects of C. asiatica hairy root lines: the expression level of centelloside biosynthetic genes, morphological traits, as well as squalene, phytosterol, and centelloside content. Three distinct categories of transformed lines were obtained: LS, harboring At-SQS; LT, overexpressing TSAR2; and LST, simultaneously carrying both transgenes. These lines displayed noticeable alterations in morphological traits, including changes in branching rate and biomass production. Furthermore, we observed that the expression of T-DNA genes, particularly aux2 and rolC genes, significantly modulated the expression of pivotal genes involved in centelloside biosynthesis. Notably, the LS lines boasted an elevated centelloside content but concurrently displayed reduced phytosterol content, a finding that underscores the intriguing antagonistic relationship between phytosterol and triterpene pathways. Additionally, the inverse correlation between the centelloside content and morphological growth values observed in LS lines was countered by the action of TSAR2 in the LST and LT lines. This difference could be attributed to the simultaneous increase in the phytosterol content in the TSAR2-expressing lines, as these compounds are closely linked to root development. Overall, these discoveries offer valuable information for the biotechnological application of C. asiatica hairy roots and their potential to increase centelloside production. Full article

Squalene is the major unsaponifiable component of virgin olive oil, the fat source of the Mediterranean diet. To evaluate its effect on the hepatic transcriptome, RNA sequencing was carried out in two groups of male Large White x Landrace pigs developing nonalcoholic steatohepatitis by feeding them a high fat/cholesterol/fructose and methionine and choline-deficient steatotic diet or the same diet with 0.5% squalene. Hepatic lipids, squalene content, steatosis, activity (ballooning + inflammation), and SAF (steatosis + activity + fibrosis) scores were analyzed. Pigs receiving the latter diet showed hepatic squalene accumulation and twelve significantly differentially expressed hepatic genes (log₂ fold change < 1.5 or <1.5) correlating in a gene network. These pigs also had lower hepatic triglycerides and lipid droplet areas and higher cellular ballooning. Glutamyl aminopeptidase (ENPEP) was correlated with triglyceride content, while alpha-fetoprotein (AFP), neutralized E3 ubiquitin protein ligase 3 (NEURL3), 2′-5′-oligoadenylate synthase-like protein (OASL), and protein phosphatase 1 regulatory inhibitor subunit 1B (PPP1R1B) were correlated with activity reflecting inflammation and ballooning, and NEURL3 with the SAF score. AFP, ENPEP, and PPP1R1B exhibited a remarkably strong discriminant power compared to those pathological parameters in both experimental groups. Moreover, the expression of PPP1R1B, TMEM45B, AFP, and ENPEP followed the same pattern in vitro using human hepatoma (HEPG2) and mouse liver 12 (AML12) cell lines incubated with squalene, indicating a direct effect of squalene on these expressions. These findings suggest that squalene accumulated in the liver is able to modulate gene expression changes that may influence the progression of non-alcoholic steatohepatitis. Full article

Trypanosomatids are mainly responsible for leishmaniasis, sleeping sickness, and Chagas disease, which are the most challenging among the neglected tropical diseases due to the problem of drug resistance. Although problems of target deconvolution and polypharmacology are encountered, a target-based approach is a rational method for screening drug candidates targeting a biomolecule that causes infections. The present study aims to summarize the latest information regarding potential inhibitors of squalene synthase and farnesyl phosphate synthase with anti-Trypanosomatidae activity. The information was obtained by referencing textbooks and major scientific databases from their inception until April 2023. Based on in vitro experiments, more than seventy compounds were reported to inhibit squalene synthase and farnesyl diphosphate synthase. Among these compounds, more than 30 were found to be active in vitro against Trypanosomatidae, inferring that these compounds can be used as scaffolds to develop new drugs against trypanosomatid-related infections. Overall, natural and synthetic products can inhibit enzymes that are crucial for the survival and virulence of trypanosomatids. Moreover, in vitro experiments have confirmed the activity of more than half of these inhibitors using cell-based assays. Nevertheless, additional studies on the cytotoxicity, pharmacokinetics, and lead optimization of potent anti-Trypanosomatid compounds should be investigated. Full article

Squalene synthase (SQS) has emerged as a promising therapeutic target for various diseases, including cancers, owing to its pivotal role in the mevalonate pathway and the antioxidant properties of squalene. Primarily, SQS orchestrates the head-to-head condensation reaction, catalyzing the fusion of two farnesyl pyrophosphate molecules, leading to the formation of squalene, which has been depicted as a highly effective oxygen-scavenging agent in in vitro studies. Recent studies have depicted this isoprenoid as a protective layer against ferroptosis due to its potential regulation of lipid peroxidation, as well as its protection against oxidative damage. Therefore, beyond its fundamental function, recent investigations have unveiled additional roles for SQS as a regulator of lipid peroxidation and programmed cell death pathways, such as ferroptosis—a type of cell death characterized by elevated levels of lipid peroxide, one of the forms of reactive oxygen species (ROS), and intracellular iron concentration. Notably, thorough explorations have shed light on the distinctive features that set SQS apart from other members within the isoprenoid synthase superfamily. Its unique biochemical structure, intricately intertwined with its reaction mechanism, has garnered significant attention. Moreover, considerable evidence substantiates the significance of SQS in various disease contexts, and its intriguing association with ferroptosis and lipid peroxidation. The objective of this report is to analyze the existing literature comprehensively, corroborating these findings, and provide an up-to-date perspective on the current understanding of SQS as a prospective therapeutic target, as well as its intricate relationship with ferroptosis. This review aims to consolidate the knowledge surrounding SQS, thereby contributing to the broader comprehension of its potential implications in disease management and therapeutic interventions. Full article

Aralia elata is an important herb due to the abundance of pentacyclic triterpenoid saponins whose important precursors are squalene and OA. Here, we found that MeJA treatment promoted both precursors accumulation, especially the latter, in transgenic A. elata, overexpressing a squalene synthase gene from Panax notoginseng(PnSS). In this study, Rhizobium-mediated transformation was used to express the PnSS gene. Gene expression analysis and high-performance liquid chromatography (HPLC) were used to identify the effect of MeJA on squalene and OA accumulation. The PnSS gene was isolated and expressed in A. elata. Transgenic lines showed a very high expression of the PnSS gene and farnesyl diphosphate synthase gene (AeFPS) and a slightly higher squalene content than the wild-type, but endogenous squalene synthase (AeSS), squalene epoxidase (AeSE), and β-amyrin synthase (Aeβ-AS) gene were decreased as well as OA content. Following one day of MeJA treatment, the expression levels of PeSS, AeSS, and AeSE genes increased significantly. On day 3, the maximum content of both products reached 17.34 and 0.70 mg·g^−1, which increased 1.39- and 4.90-fold than in the same lines without treatment. Transgenic lines expressing PnSS gene had a limited capability to promote squalene and OA accumulation. MeJA strongly activated their biosynthesis pathways, leading to enhance yield. Full article