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1.
J Ethnopharmacol ; 334: 118542, 2024 Nov 15.
Artículo en Inglés | MEDLINE | ID: mdl-38992404

RESUMEN

ETHNOPHARMACOLOGICAL RELEVANCE: Dried roots of Peucedanum decursivum, a traditional Chinese medicine (TCM), has historically respiratory diseases such as cough, thick phlegm, headache, fever, and gynecological diseases, rheumatoid arthritis, and nasopharyngeal carcinoma. AIM OF THE STUDY: Made an endeavor to evaluate the research trajectory of P. decursivum, comprehensively discern its developmental status, and offer a guideline for future investigations. MATERIALS AND METHODS: A meticulous search of literatures and books from 1955 to 2024 via databases like PubMed, Web of Science and CNKI was conducted, including topics and keywords of " P. decursivum" "Angelica decursivum" and "Zihua Qianhu". RESULTS: P. decursivum and its prescriptions have traditionally been used for treating phlegm-heat cough, wind-heat cough, gastrointestinal diseases, pain relief and so on. It contains 234 identified compounds, encompassing coumarins, terpenes, volatile oils, phenolic acids, fatty acids and derivatives. It exhibits diverse pharmacological activities, including anti-asthmatic, anti-inflammatory, antioxidant effects, anti-hypertensive, anti-diabetic, anti-Alzheimer, and anti-cancer properties, primarily attributed to coumarins. Microscopic identification, HPLC fingerprinting, and bioinformatics identification are the primary methods currently used for the quality control. CONCLUSION: P. decursivum demonstrates anti-asthmatic, anti-inflammatory, and antioxidant effects, aligning with its traditional use. However, experimental validation of its efficacy against phlegm and viruses is needed. Additionally, analgesic effects mentioned in historical texts lack modern pharmacological studies. Numerous isolated compounds exhibit highly valuable medicinal properties. Future research can delve into exploring these substances further. Rigorous of heavy metal contamination, particularly Cd and Pb, is necessary. Simultaneously, investigating its pharmacokinetics and toxicity in humans is crucial for the safety.


Asunto(s)
Apiaceae , Etnobotánica , Etnofarmacología , Fitoquímicos , Control de Calidad , Humanos , Fitoquímicos/farmacología , Fitoquímicos/química , Fitoquímicos/uso terapéutico , Apiaceae/química , Animales , Medicamentos Herbarios Chinos/farmacología , Medicamentos Herbarios Chinos/química , Medicamentos Herbarios Chinos/uso terapéutico , Medicina Tradicional China/métodos
2.
Genes (Basel) ; 15(6)2024 Jun 12.
Artículo en Inglés | MEDLINE | ID: mdl-38927707

RESUMEN

Phenylalanine ammonia lyase (PAL) is a key enzyme regulating the biosynthesis of the compounds of the phenylpropanoid pathway. This study aimed to isolate and characterize PAL genes from Ferula pseudalliacea Rech.f. (Apiales: Apiaceae) to better understand the regulation of metabolite production. Three PAL gene isoforms (FpPAL1-3) were identified and cloned using the 3'-RACE technique and confirmed by sequencing. Bioinformatics analysis revealed important structural features, such as phosphorylation sites, physicochemical properties, and evolutionary relationships. Expression analysis by qPCR demonstrated the differential transcription profiles of each FpPAL isoform across roots, stems, leaves, flowers, and seeds. FpPAL1 showed the highest expression in stems, FpPAL2 in roots and flowers, and FpPAL3 in flowers. The presence of three isoforms of PAL in F. pseudalliacea, along with the diversity of PAL genes and their tissue-specific expression profiles, suggests that complex modes of regulation exist for phenylpropanoid biosynthesis in this important medicinal plant. The predicted interaction network revealed associations with key metabolic pathways, emphasizing the multifaceted roles of these PAL genes. In silico biochemical analyses revealed the hydrophilicity of the FpPAL isozyme; however, further analysis of substrate specificity and enzyme kinetics can clarify the specific role of each FpPAL isozyme. These comprehensive results increase the understanding of PAL genes in F. pseudalliacea, helping to characterize their contributions to secondary metabolite biosynthesis.


Asunto(s)
Ferula , Regulación de la Expresión Génica de las Plantas , Fenilanina Amoníaco-Liasa , Proteínas de Plantas , Fenilanina Amoníaco-Liasa/genética , Fenilanina Amoníaco-Liasa/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Ferula/genética , Ferula/metabolismo , Filogenia , Flores/genética
3.
Dev Cell ; 59(11): 1363-1378.e4, 2024 Jun 03.
Artículo en Inglés | MEDLINE | ID: mdl-38579719

RESUMEN

The mechanism underlying the ability of rice to germinate underwater is a largely enigmatic but key research question highly relevant to rice cultivation. Moreover, although rice is known to accumulate salicylic acid (SA), SA biosynthesis is poorly defined, and its role in underwater germination is unknown. It is also unclear whether peroxisomes, organelles essential to oilseed germination and rice SA accumulation, play a role in rice germination. Here, we show that submerged imbibition of rice seeds induces SA accumulation to promote germination in submergence. Two submergence-induced peroxisomal Oryza sativa cinnamate:CoA ligases (OsCNLs) are required for this SA accumulation. SA exerts this germination-promoting function by inducing indole-acetic acid (IAA) catabolism through the IAA-amino acid conjugating enzyme GH3. The metabolic cascade we identified may potentially be adopted in agriculture to improve the underwater germination of submergence-intolerant rice varieties. SA pretreatment is also a promising strategy to improve submerged rice germination in the field.


Asunto(s)
Germinación , Oryza , Peroxisomas , Reguladores del Crecimiento de las Plantas , Proteínas de Plantas , Oryza/metabolismo , Oryza/crecimiento & desarrollo , Germinación/fisiología , Peroxisomas/metabolismo , Reguladores del Crecimiento de las Plantas/metabolismo , Proteínas de Plantas/metabolismo , Regulación de la Expresión Génica de las Plantas , Coenzima A Ligasas/metabolismo , Ácidos Indolacéticos/metabolismo , Semillas/metabolismo , Semillas/crecimiento & desarrollo , Ácido Salicílico/metabolismo , Cinamatos/metabolismo
4.
Front Mol Biosci ; 11: 1341950, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38516193

RESUMEN

Introduction: Parkinson's disease (PD) is the most common motor neurodegenerative disease worldwide. Given the complexity of PD etiology and the different metabolic derangements correlated to the disease, metabolomics profiling of patients is a helpful tool to identify patho-mechanistic pathways for the disease development. Dopamine metabolism has been the target of several previous studies, of which some have reported lower phenylalanine and tyrosine levels in PD patients compared to controls. Methods: In this study, we have collected plasma from 27 PD patients, 18 reference controls, and 8 high-risk controls to perform a metabolomic study using liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS). Results: Our findings revealed higher intensities of trans-cinnamate, a phenylalanine metabolite, in patients compared to reference controls. Thus, we hypothesize that phenylalanine metabolism has been shifted to produce trans-cinnamate via L-phenylalanine ammonia lyase (PAL), instead of producing tyrosine, a dopamine precursor, via phenylalanine hydroxylase (PAH). Discussion: Given that these metabolites are precursors to several other metabolic pathways, the intensities of many metabolites such as dopamine, norepinephrine, and 3-hydroxyanthranilic acid, which connects phenylalanine metabolism to that of tryptophan, have been altered. Consequently, and in respect to Metabolic Control Analysis (MCA) theory, the levels of tryptophan metabolites have also been altered. Some of these metabolites are tryptamine, melatonin, and nicotinamide. Thus, we assume that these alterations could contribute to the dopaminergic, adrenergic, and serotonergic neurodegeneration that happen in the disease.

5.
Angew Chem Int Ed Engl ; 63(20): e202400955, 2024 May 13.
Artículo en Inglés | MEDLINE | ID: mdl-38489506

RESUMEN

Vitrimers represent an emerging class of polymeric materials that combine the desirable characteristics of both thermoplastics and thermosets achieved through the design of dynamic covalent bonds within the polymer networks. However, these materials are prone to creep due to the inherent instability of dynamic covalent bonds. Consequently, there are pressing demands for the development of robust and stable dynamic covalent chemistries. Here, we report a catalyst-free α-acetyl cinnamate/acetoacetate (α-AC/A) exchange reaction to develop vitrimers with remarkable creep resistance. Small-molecule model studies revealed that the α-AC/A exchange occurred at temperatures above 140 °C in bulk, whereas at 120 °C, this reaction was absent. For demonstration in the case of polymers, copolymers derived from common vinyl monomers were crosslinked with terephthalaldehyde to produce α-AC/A vitrimers with tunable thermal and mechanical performance. All resulting α-AC/A vitrimers exhibited high stability, especially in terms of creep resistance at 120 °C, while retaining commendable reprocessability when subjected to high temperatures. This work showcases the α-AC/A exchange reaction as a novel and robust dynamic covalent chemistry capable of imparting both reprocessability and high stability to cross-linked networks.

6.
Diabetes Metab Syndr Obes ; 17: 1183-1197, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38469107

RESUMEN

Background: AMP-activated protein kinase (AMPK) plays a critical role in energy metabolism. Its activation leads to the phosphorylation of downstream proteins such as acetyl-CoA carboxylase (ACC) and sterol regulatory element-binding protein-1 (SREBP1), subsequently inhibiting de novo fatty acid synthesis, thereby reducing intracellular triglyceride accumulation. MC is a compound found in extracts from Zanthoxylum armatum DC plants. Research has shown that MC can inhibit the differentiation of 3T3-L1 adipocytes through the CAMKK2-AMPK pathway. However, the biological effect of MC in HepG2 cells remains unknown. Methods: In this study, we utilized HepG2 cells to establish a model of MAFLD through FFAs stimulation. We investigated the biological effects of MC on HepG2 cells and studied its impact on lipid metabolism. Small interfering RNA was employed to explore the mechanism by which MC activates AMPK. Finally, molecular docking was conducted, establishing a model of the interaction between AMPK and MC. Results: We observed that MC can alleviate triglyceride accumulation in HepG2 cells. We observed the elevated p-AMPK/AMPK, P-ACC/ ACC, and elevated CPT1a after treatment of MC in HepG2 cells. The interference of CAMKK2 mRNA did not impact the ability of MC to phosphorylate AMPK. Compound C attenuates the ability of MC to increase p-AMPK. Molecular docking results led us to hypothesize that MC directly interacts with AMPK, resulting in AMPK phosphorylation and improved lipid accumulation in HepG2 cells.

7.
J Ethnopharmacol ; 326: 117913, 2024 May 23.
Artículo en Inglés | MEDLINE | ID: mdl-38360380

RESUMEN

ETHNOPHARMACOLOGICAL RELEVANCE: Kaempferia galanga Linn. is an aromatic medicinal herb with extensively applied in India, China, Malaysia and other South Asia countries for thousands of years. It has been mentioned to treat abdominal tumors. Ethyl cinnamate (EC), one of the main chemical constituents of the rhizome of K. galanga, exhibited nematocidal, sedative and vasorelaxant activities. However, its anti-angiogenic activity, and anti-tumor effect have not been investigated. AIM OF THE STUDY: To investigate the anti-angiogenic mechanism of EC and its anti-tumor effect by suppressing angiogenesis. MATERIALS AND METHODS: The in vitro anti-angiogenic effect was evaluated using HUVECs model induced by VEGF and zebrafish model in vivo. The influence of the EC on phosphorylation of VEGFR2 and its downstream signaling pathways were evaluated by western blotting assay. Molecule docking technology was conducted to explore the interaction between EC and VEGFR2. SPR assay was used for detecting the binding affinity between EC and VEGFR2. To further investigate the molecular mechanism of EC on anti-angiogenesis, VEGFR2 knockdown in HUVECs and examined the influence of the EC. Anti-tumor activity of EC was evaluated using colony formation assay and apoptosis assay. The inhibitory effect of EC on tumor growth was explored using HT29 colon cancer xenograft model. RESULTS: EC obviously inhibited proliferation, migration, invasion and tube formation of VEGF-induced HUVECs. EC also induced apoptosis of HUVECs. Moreover, it inhibited the development of vessel formation in zebrafish. Further investigations demonstrated that EC could suppress the phosphorylation of VEGFR2, and its downstream signaling pathways were altered in VEGF-induced HUVECs. EC formed a hydrogen bond to bind with the ATP binding site of the VEGFR2, and EC-VEGFR2 interaction was shown in SPR assay. The suppressive effect of EC on angiogenesis was abrogated after VEGFR2 knockdown in HUVECs. EC inhibited the colon cancer cells colony formation and induced apoptosis. In addition, EC suppressed tumor growth in colon cancer xenograft model, and no detectable hepatotoxicity and nephrotoxicity. In addition, it inhibited the phosphorylation of VEGFR2, and its downstream signal pathways in tumor. CONCLUSIONS: EC could inhibit tumor growth in colon cancer by suppressing angiogenesis via VEGFR2 signaling pathway, and suggested EC as a promising candidate for colon cancer treatment.


Asunto(s)
Cinamatos , Neoplasias del Colon , Neoplasias Colorrectales , Animales , Humanos , Pez Cebra , Células Endoteliales de la Vena Umbilical Humana , Factor A de Crecimiento Endotelial Vascular/metabolismo , Proliferación Celular , Movimiento Celular , Transducción de Señal , Inhibidores de la Angiogénesis/farmacología , Inhibidores de la Angiogénesis/uso terapéutico , Neoplasias Colorrectales/metabolismo , Neoplasias del Colon/tratamiento farmacológico , Receptor 2 de Factores de Crecimiento Endotelial Vascular/metabolismo , Neovascularización Patológica/metabolismo
8.
Plant Sci ; 341: 111989, 2024 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-38232819

RESUMEN

Hornworts, as the sister group to liverworts and mosses, comprise bryophytes, which are critical in understanding the evolution of key land plant traits. Cinnamate 4-hydroxylase (C4H) catalyzes the second step of the phenylpropanoid pathway to synthesize the precursor of numerous phenolic compounds, such as lignin and flavonoids. However, C4H in the hornwort Anthoceros angustus has not yet been cloned and functionally characterized. In this work, we screened the transcriptome database of A. angustus and identified one C4H gene, AnanC4H. AnanC4H maintained conserved cytochrome P450 domains with other typical plant C4Hs. Ultraviolet B irradiation and exogenous application of methyl jasmonate (MeJA) induced the expression of AnanC4H to varying degrees. The coding sequence of AnanC4H was expressed in yeast, and the recombinant proteins were isolated. The recombinant proteins of AnanC4H catalyzed the conversion of trans-cinnamic acid to p-coumaric acid and catalyzed the conversion of 3-hydroxycinnamic acid to caffeic acid. AnanC4H showed higher affinity for trans-cinnamic acid than for 3-hydroxycinnamic acid, but there was no significant difference in the catalytic efficiency of AnanC4H for the two substrates in vitro. Moreover, the expression of AnanC4H in Arabidopsis thaliana led to an increase in both the lignin content and the number of lignified cells in stems. However, there was no significant change in flavonoid content in transgenic Arabidopsis plants.


Asunto(s)
Anthocerotophyta , Arabidopsis , Cinamatos , Transcinamato 4-Monooxigenasa/genética , Transcinamato 4-Monooxigenasa/metabolismo , Anthocerotophyta/genética , Anthocerotophyta/metabolismo , Ácidos Cumáricos , Lignina/metabolismo , Saccharomyces cerevisiae/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Clonación Molecular , Proteínas Recombinantes/genética
9.
Biotechnol J ; 19(1): e2300275, 2024 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-37861236

RESUMEN

Phenylalanine ammonia-lyase (PAL) is a crucial enzyme for various biotechnology applications, such as producing phenols, antioxidants, and nutraceuticals. However, feedback inhibition from its product, cinnamic acid, limits its forward reaction rate. Therefore, this study aims to address the feedback inhibition in PAL using enzyme engineering strategies. Random and site-directed mutagenesis approaches were utilized to screen mutant enzymes with ameliorated tolerance against cinnamic acid. A thermotolerant and cinnamate-tolerant mutant was rationally identified using a high throughput screening method and subsequent biochemical characterization. We evaluated cinnamate affinity among the seven rationally selected mutations, and the T102E mutation was identified as the most promising mutant. This mutant showed a six-fold reduction in the affinity of PAL for cinnamic acid and a two-fold increase in operational stability compared with native PAL. Furthermore, the enzyme was immobilized on carbon nanotubes to increase its robustness and reusability. The immobilized mutant PAL showed greater efficiency in the deamination of phenylalanine present in protein hydrolysate than its free form. The rationale behind the enhancement of cinnamate tolerance was validated using molecular dynamic simulations. Overall, the knowledge of the sequence-function relationship of PAL was applied to drive enzyme engineering to develop highly tolerant PAL.


Asunto(s)
Nanotubos de Carbono , Fenilanina Amoníaco-Liasa , Fenilanina Amoníaco-Liasa/genética , Fenilanina Amoníaco-Liasa/química , Fenilanina Amoníaco-Liasa/metabolismo , Retroalimentación , Cinamatos , Biotransformación
10.
Appl Microbiol Biotechnol ; 107(24): 7581-7599, 2023 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-37801099

RESUMEN

Resveratrol (RES) is a secondary metabolite synthesized by plants in response to environmental stress and pathogen infection, which is of great significance for the industrial production of RES by fermentation culture. In this study, we aimed to explore the biosynthesis pathway of RES and its key enzymes in the Priestia megaterium PH3, which was isolated and screened from peanut fruit. Through Liquid Chromatography-Mass Spectrometry (LC-MS) analysis, we quantified the RES content and distribution in the culture medium and determined that Priestia megaterium PH3 mainly secreted RES extracellularly. Furthermore, the highest production of RES was observed in YPD, yielding an impressive 127.46 ± 6.11 µg/L. By optimizing the fermentation conditions, we achieved a remarkable RES yield of 946.82 ± 24.74 µg/L within just 2 days, which represents the highest reported yield for a natural isolate produced in such a short time frame. Our investigation revealed that the phenylpropane pathway is responsible for RES synthesis in this bacterium, with cinnamate 4-hydroxylase (C4H) identified as the main rate-limiting enzyme. Overall, our findings highlight the robust RES production capabilities of Priestia megaterium PH3, offering novel insights and potential applications for bacterial fermentation in RES production. KEY POINTS: • RES synthesized by the bacterium was confirmed through the phenylpropane pathway. • The key rate-limiting enzyme for biosynthesis-RES is C4H. • RES reached 946.82 ± 24.74 µg/L after fermentation for 2 days.


Asunto(s)
Bacillus megaterium , Resveratrol/metabolismo , Fermentación , Espectrometría de Masas , Bacillus megaterium/metabolismo , Metabolismo Secundario
11.
Acta Biochim Biophys Sin (Shanghai) ; 55(11): 1806-1818, 2023 11 25.
Artículo en Inglés | MEDLINE | ID: mdl-37654075

RESUMEN

Effective and non-toxic therapeutic agents are lacking for the prevention and treatment of colitis. Previous studies found that methyl cinnamate (MC), extracted from galangal ( Alpinia officinarum Hance), has anti-inflammatory properties. However, whether MC is effective as anti-colitis therapy remains unknown. In this study, we investigate the therapeutic effects of MC on dextran sulfate sodium (DSS)-induced colitis in mice and further explore its potential mechanism of action. MC treatment relieves symptoms associated with DSS-induced colitis, including the recovery of DSS-induced weight loss, decreases the disease activity index score, and increases the colon length without toxic side effects. MC treatment protects the integrity of the intestinal barrier in mice with DSS-induced colitis and inhibits the overexpression of pro-inflammatory cytokines in vivo and in vitro. Moreover, the MAPK signaling pathway is found to be closely related to the treatment with MC of colitis. Western blot analysis show that phosphorylation of the p38 protein in colon tissues treated with MC is markedly reduced and phosphorylation levels of the p38, JNK and ERK proteins are significantly decreased in RAW 264.7 cells treated with MC, indicating that the mechanism of MC in treating DSS-induced colitis could be achieved by inhibiting the MAPK signaling pathway. Furthermore, 16S RNA sequencing analysis show that MC can improve intestinal microbial dysbiosis in mice with DSS-induced colitis. Altogether, these findings suggest that MC may be a novel therapeutic candidate with anti-colitis efficacy. Furthermore, MC treatment relieves the symptoms of colitis by inhibiting the MAPK signaling pathway and improving the intestinal microbiota.


Asunto(s)
Colitis , Ratones , Animales , Sulfato de Dextran/toxicidad , Ratones Endogámicos C57BL , Colitis/inducido químicamente , Colitis/tratamiento farmacológico , Colitis/prevención & control , Transducción de Señal , Colon/metabolismo , Modelos Animales de Enfermedad
12.
Int J Mol Sci ; 24(15)2023 Jul 29.
Artículo en Inglés | MEDLINE | ID: mdl-37569517

RESUMEN

The Aedes aegypti mosquito is a vector of severe diseases with high morbidity and mortality rates. The most commonly used industrial larvicides have considerable toxicity for non-target organisms. This study aimed to develop and evaluate liquid and solid carrier systems to use pentyl cinnamate (PC), derived from natural sources, to control Ae. aegypti larvae. The liquid systems consisting of nanoemulsions with different lecithins systems were obtained and evaluated for stability over 30 days. Microparticles (MPs) were obtained by the spray drying of the nanoemulsions using maltodextrin as an adjuvant. Thermal, NMR and FTIR analysis indicated the presence of PC in microparticles. Indeed, the best nanoemulsion system was also the most stable and generated the highest MP yield. The PC larvicidal activity was increased in the PC nanoemulsion system. Therefore, it was possible to develop, characterize and obtain PC carrier systems active against Ae. aegypti larvae.


Asunto(s)
Aedes , Insecticidas , Animales , Insecticidas/química , Mosquitos Vectores , Cinamatos/farmacología , Larva
13.
J Agric Food Chem ; 71(28): 10590-10597, 2023 Jul 19.
Artículo en Inglés | MEDLINE | ID: mdl-37399340

RESUMEN

Sumatra benzoin, a resin produced by Styrax benzoin and Styrax paralleloneurum, is used as an aromatic agent and may have the potential to be developed as a new agricultural fungicide. In this context, we performed a comprehensive metabolite profiling of a commercial grade A resin by high-performance liquid chromatography coupled with photodiode array detection, evaporative light scattering detection, and mass spectrometry (HPLC-PDA-ELSD-MS) analysis in combination with 1H NMR. Thirteen compounds including a new cinnamic acid ester containing two p-coumaroyl residues were identified after preparative isolation. These compounds accounted for an estimated 90% of the crude resin according to 1H NMR analysis. The two major constituents, p-coumaryl cinnamate (5) and sumaresinolic acid (11), were quantified by HPLC analysis. In a next step, the chemical profiles and the content in p-coumaryl cinnamate were compared in a large set of resin samples of different quality grades that were obtained from various commercial suppliers in Sumatra. The qualitative profiles of the samples were very similar, but significant quantitative differences were observed between different quality grades and origins of the samples for the relative contents.


Asunto(s)
Benzoína , Styrax , Styrax/química , Indonesia , Espectroscopía de Resonancia Magnética , Cinamatos , Cromatografía Líquida de Alta Presión/métodos
14.
Molecules ; 28(6)2023 Mar 21.
Artículo en Inglés | MEDLINE | ID: mdl-36985814

RESUMEN

Leishmania infantum is the etiological agent of visceral leishmaniasis (VL) in South America, the Mediterranean basin, and West and Central Asia. The most affected country, Brazil, reported 4297 VL cases in 2017. L. infantum is transmitted by female phlebotomine sand flies during successive blood meals. There are no validated vaccines to prevent the infection and the treatment relies on drugs that often present severe side effects, which justify the efforts to find new antileishmanial drugs. Cinnamic acid derivatives have shown several pharmacological activities, including antiparasitic action. Therefore, in the present study, the biological evaluation of cinnamic acid and thirty-four derivatives against L. infantum is reported. The compounds were prepared by several synthesis methods and characterized by spectroscopic techniques and high-resolution mass spectrometry. The results revealed that compound 32 (N-(4-isopropylbenzyl)cinnamamide) was the most potent antileishmanial agent (IC50 = 33.71 µM) with the highest selectivity index (SI > 42.46), followed by compound 15 (piperonyl cinnamate) with an IC50 = 42.80 µM and SI > 32.86. Compound 32 was slightly less potent and nineteen times more selective for the parasite than amphotericin B (MIC = 3.14 uM; SI = 2.24). In the molecular docking study, the most likely target for the compound in L. infantum was aspartyl aminopeptidase, followed by aldehyde dehydrogenase, mitochondrial. The data obtained show the antileishmanial potential of this class of compounds and may be used in the search for new drug candidates against Leishmania species.


Asunto(s)
Antiprotozoarios , Leishmania infantum , Leishmaniasis Visceral , Femenino , Humanos , Simulación del Acoplamiento Molecular , Antiprotozoarios/química , Leishmaniasis Visceral/tratamiento farmacológico , Cinamatos/farmacología , Cinamatos/uso terapéutico , Brasil
15.
Int J Mol Sci ; 24(6)2023 Mar 11.
Artículo en Inglés | MEDLINE | ID: mdl-36982470

RESUMEN

C4H (cinnamate 4-hydroxylase) is a pivotal gene in the phenylpropanoid pathway, which is involved in the regulation of flavonoids and lignin biosynthesis of plants. However, the molecular mechanism of C4H-induced antioxidant activity in safflower still remains to be elucidated. In this study, a CtC4H1 gene was identified from safflower with combined analysis of transcriptome and functional characterization, regulating flavonoid biosynthesis and antioxidant defense system under drought stress in Arabidopsis. The expression level of CtC4H1 was shown to be differentially regulated in response to abiotic stresses; however, a significant increase was observed under drought exposure. The interaction between CtC4H1 and CtPAL1 was detected using a yeast two-hybrid assay and then verified using a bimolecular fluorescence complementation (BiFC) analysis. Phenotypic and statistical analysis of CtC4H1 overexpressed Arabidopsis demonstrated slightly wider leaves, long and early stem development as well as an increased level of total metabolite and anthocyanin contents. These findings imply that CtC4H1 may regulate plant development and defense systems in transgenic plants via specialized metabolism. Furthermore, transgenic Arabidopsis lines overexpressing CtC4H1 exhibited increased antioxidant activity as confirmed using a visible phenotype and different physiological indicators. In addition, the low accumulation of reactive oxygen species (ROS) in transgenic Arabidopsis exposed to drought conditions has confirmed the reduction of oxidative damage by stimulating the antioxidant defensive system, resulting in osmotic balance. Together, these findings have provided crucial insights into the functional role of CtC4H1 in regulating flavonoid biosynthesis and antioxidant defense system in safflower.


Asunto(s)
Arabidopsis , Carthamus tinctorius , Arabidopsis/metabolismo , Antioxidantes/metabolismo , Flavonoides/metabolismo , Carthamus tinctorius/genética , Cinamatos/metabolismo , Estrés Fisiológico , Regulación de la Expresión Génica de las Plantas , Plantas Modificadas Genéticamente/genética , Sequías , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo
16.
Methods Mol Biol ; 2562: 123-133, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-36272071

RESUMEN

Tissue clearing turns otherwise turbid and opaque tissue transparent, enabling imaging deep within tissues. The nontransparent nature of most tissues is due to the refractive index mismatch between its three major constituent components (lipids, proteins, and water). All tissue clearing methods rectify this mismatch by homogenizing the refractive index within the tissue and carefully matching it to the surrounding media. Here we describe a detailed protocol to clear a wide range of salamander tissues. We also include several optional steps such as depigmentation, antibody staining, and tissue mounting. These steps are optional, and do not change anything in the steps needed for tissue clearing. Depending on the fluorescent signal and optics employed, images up to several millimeters inside of the tissue can be acquired.


Asunto(s)
Cinamatos , Lípidos , Coloración y Etiquetado , Agua , Imagenología Tridimensional/métodos
17.
Molecules ; 27(21)2022 Oct 22.
Artículo en Inglés | MEDLINE | ID: mdl-36363974

RESUMEN

Cinnamic acid is one of the phenolic compounds that is isolated from cinnamon, or other natural plants, and has a wide range of physiological activities. However, the application of cinnamic acid is limited due to its poor solubility and low oral bioavailability. In this study, the feasibility of producing octyl cinnamate by ultrasonic assistance, combined with a rotary evaporation under vacuum, was studied using methyl cinnamate and octanol as the starting materials. A Box-Behnken design (BBD) was employed to evaluate the effects of the operation parameters, including reaction temperature (55-75 °C), reaction time (4-12 h), and ultrasonic power (90-150 W) on the production of octyl cinnamate. Meanwhile, the synthesis process was further optimized by the modeling response surface methodology (RSM). The data indicated that octyl cinnamate was efficiently synthesized from methyl cinnamate and octanol using the ultrasound plus vacuum system; further, this system was superior to the conventional method. According to the RSM model for the actual experiments, a reaction temperature of 74.6 °C, a reaction time of 11.1 h, and an ultrasound power of 150 W were determined to be the best conditions for the maximum molar conversion of octyl cinnamate (93.8%). In conclusion, the highly efficient synthesis of octyl cinnamate by a rotary evaporator with an ultrasound plus vacuum system was achieved via RSM optimization.


Asunto(s)
Cinamatos , Vacio , Octanoles
18.
Pharm Biol ; 60(1): 1969-1980, 2022 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-36226757

RESUMEN

CONTEXT: Some studies reported the chemical content and antimicrobial properties of Ocimum basilicum L. (Lamiaceae), relevant to the ecological variations in some areas of Egypt and other countries, yet no research was conducted on the plant cultivated in the central delta region of Egypt. Also, no previous data reported on inhibition of ß-lactamases by O. basilicum. OBJECTIVE: To assess ß-lactamases inhibition by O. basilicum extracts and the individual constituents. MATERIALS AND METHODS: Dried aerial parts of O. basilicum were extracted by hydrodistillation for preparation of essential oil and by methanol for non-volatile constituents. Essential oil content and the methanol extract were analysed by GC-MS and UPLC-PDA-MS/MS, respectively. Methyl cinnamate was isolated and analysed by NMR. Broth microdilution method was used to investigate the antimicrobial against resistant clinical isolates of Escherichia coli identified by double disc synergy, combination disc tests and PCR. The most active oil content was further tested with a nitrocefin kit for ß-lactamase inhibition and investigated by docking. RESULTS: O. basilicum was found to contain methyl cinnamate as the major content of the essential oil. More interestingly, methyl cinnamate inhibited ESBL ß-lactamases of the type CTX-M. The in vitro IC50 using nitrocefin kit was 11.6 µg/mL vs. 8.1 µg/mL for clavulanic acid as a standard ß-lactamase inhibitor. DISCUSSION AND CONCLUSIONS: This is the first study to report the inhibitory activity of O. basilicum oil and methyl cinnamate against ß-lactamase-producing bacteria. The results indicate that methyl cinnamate could be a potential alternative for ß-lactamase inhibition.


Asunto(s)
Lamiaceae , Ocimum basilicum , Aceites Volátiles , Antibacterianos/farmacología , Cefalosporinas , Cinamatos , Ácido Clavulánico , Egipto , Metanol , Ocimum basilicum/química , Fitoquímicos/farmacología , Extractos Vegetales/farmacología , Espectrometría de Masas en Tándem , Inhibidores de beta-Lactamasas/farmacología , beta-Lactamasas
19.
Gels ; 8(10)2022 Oct 09.
Artículo en Inglés | MEDLINE | ID: mdl-36286140

RESUMEN

Long-term UV radiation exposure can induce skin disorders such as cancer and photoallergic reactions. Natural products have been considered as non-irritate and potential sunscreen resources due to their UV absorption and anti-inflammatory activities. This study aimed to evaluate the in vitro ultraviolet radiation protective effect and anti-inflammatory activity of K. galanga rhizome oil and microemulsions. The chemical components of K. galanga rhizome oil was analyzed via gas chromatography coupled with mass spectrometry. Microemulsions containing K. galanga rhizome oil were formulated using a phase-titration method. The microemulsion was characterized for droplet size, polydispersity index, and zeta potential, using a dynamic light-scattering technique. The physical and chemical stability of the microemulsion were evaluated via a dynamic light scattering technique and UV-Vis spectrophotometry, respectively. The UV protection of K. galanga rhizome oil and its microemulsion were investigated using an ultraviolet transmittance analyzer. The protective effect of K. galanga rhizome oil against LPS-induced inflammation was investigated via MTT and nitric oxide inhibitory assays. In addition, a hydrogel containing K. galanga rhizome oil microemulsion was developed, stored for 90 days at 4, 30, and 45 °C, and characterized for viscosity, rheology, and pH. The chemical degradation of the main active compound in the microemulsion was analyzed via UV-Vis spectrophotometry. The formulated O/W microemulsion contained a high loading efficiency (101.24 ± 2.08%) of K. galanga rhizome oil, suggesting a successful delivery system of the oil. The size, polydispersity index, and zeta potential values of the microemulsion were optimized and found to be stable when stored at 4, 30, and 45 °C. K. galanga rhizome oil and microemulsion demonstrated moderate sun protective activity and reduced the nitric oxide production induced by LPS in macrophage cells, indicating that microemulsion containing K. galanga rhizome oil may help protect human skin from UV damage and inflammation. A hydrogel containing K. galanga rhizome oil microemulsion was developed as a topical preparation. The hydrogel showed good physical stability after heating and cooling cycles and long-term storage (3 months) at 4 °C. The use of K. galanga rhizome oil as a natural sun-protective substance may provide a protective effect against inflammation on the skin. K. galanga rhizome oil microemulsion was successfully incorporated into the hydrogel and has the potential to be used as a topical sunscreen preparation.

20.
J Agric Food Chem ; 70(25): 7736-7741, 2022 Jun 29.
Artículo en Inglés | MEDLINE | ID: mdl-35709502

RESUMEN

Methyl cinnamate with a fruity balsamic odor is an important fragrance ingredient in perfumes and cosmetics. Chemical processes are currently the only means of producing methyl cinnamate. But consumers prefer natural flavors. Therefore, it is necessary to design and develop microbial cell factories for the production of methyl cinnamate. In this study, we established for the first time a biosynthetic pathway in engineered Escherichia coli for production of methyl cinnamate from glucose. We further increased the methyl cinnamate production to 302 mg/L by increasing the availability of the metabolic precursors. Finally, the titer was increased to 458 mg/L in a two-phase culture system.


Asunto(s)
Escherichia coli , Ingeniería Metabólica , Vías Biosintéticas , Cinamatos/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo
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