Propyl (E)-3-(furan-2-yl) Acrylate: a synthetic antifungal potential with a regulatory effect on the biosynthesis of ergosterol in Candida Albicans

Abstract The genus Candida represents the main cause of infections of fungal origin. Some species stand out as disease promoters in humans, such as C. albicans, C. glabrata, C. parapsilosis, and C. tropicalis. This study evaluated the antifungal effects of propyl (E)-3-(furan-2-yl) acrylate. The minimum inhibitory concentration of the synthetic compound, amphotericin B and fluconazole alone against four species of Candida ranged from 64 to 512 µg/mL, 1 to 2 µg/mL, and 32 to 256 µg/mL, respectively. The synergistic effect of the test substance was observed when associated with fluconazole against C. glabrata, there was no antagonism between the substances against any of the tested strains. The potential drug promoted morphological changes in C. albicans, decreasing the amount of resistance, virulence, and reproduction structures, such as the formation of pseudohyphae, blastoconidia, and chlamydospores, ensuring the antifungal potential of this substance. It was also possible to identify the fungicidal profile of the test substance through the study of the growth kinetics of C. albicans. Finally, it was observed that the test compound inhibited the ergosterol biosynthesis by yeast

Controlling antifungal activity with light: Optical regulation of fungal ergosterol biosynthetic pathway with photo-responsive CYP51 inhibitors

Invasive fungal infections (IFIs) have been associated with high mortality, highlighting the urgent need for developing novel antifungal strategies. Herein the first light-responsive antifungal agents were designed by optical control of fungal ergosterol biosynthesis pathway with photocaged triazole lanosterol 14α-demethylase (CYP51) inhibitors. The photocaged triazoles completely shielded the CYP51 inhibition. The content of ergosterol in fungi before photoactivation and after photoactivation was 4.4% and 83.7%, respectively. Importantly, the shielded antifungal activity (MIC80 ≥ 64 μg/mL) could be efficiently recovered (MIC80 = 0.5-8 μg/mL) by light irradiation. The new chemical tools enable optical control of fungal growth arrest, morphological conversion and biofilm formation. The ability for high-precision antifungal treatment was validated by in vivo models. The light-activated compound A1 was comparable to fluconazole in prolonging survival in Galleria mellonella larvae with a median survival of 14 days and reducing fungal burden in the mouse skin infection model. Overall, this study paves the way for precise regulation of antifungal therapy with improved efficacy and safety.

Mitochondrial dysfunction on Leishmania (Leishmania) amazonensis induced by ketoconazole: insights into drug mode of action

BACKGROUND Leishmania parasites cause leishmaniasis that range from self-limiting cutaneous lesions to more serious forms of the disease. The search for potential drug targets focusing on biochemical and metabolic pathways revealed the sterol biosynthesis inhibitors (SBIs) as a promising approach. In this class of inhibitors is found ketoconazole, a classical inhibitor of 14α-methysterol 14-demethylase. OBJECTIVE The present study aimed to better understand the biological response of Leishmania (Leishmania) amazonensis promastigotes at the cellular level after ketoconazole treatment. METHODS Herein, techniques, such as fluorimetry, flow cytometry, fluorescence microscopy, electron and scanning microscopy were used to investigate the cellular structures and to identify organelles affected by ketoconazole treatment. FINDINGS The study demonstrated, for the first time, the effect of ketoconazole on mitochondrion functioning and its probable relationship to cell cycle and death on L. (L.) amazonensis promastigotes (IFLA/BR/67/PH8 strain). MAIN CONCLUSIONS Ketoconazole-induced mitochondrial damages led to hyperpolarisation of this single organelle and autophagic vacuoles formation, as a parasite survival strategy. These damages did not reflect directly on the parasite cell cycle, but drove the parasites to death, making them susceptible to ketoconazole treatment in in vitro models.

Steroids from Diplazium esculentum: Antiplasmodial activity and molecular docking studies to investigate their binding modes

@#Diplazium esculentum is an edible fern commonly consumed by the local community in Malaysia either as food or medicine. Isolation work on the ethyl acetate extract of the stem of D. esculentum resulted in the purification of two steroids, subsequently identified as stigmasterol (compound 1) and ergosterol5,8-endoperoxide (compound 2). Upon further testing, compound 2 displayed strong inhibitory activity against the Plasmodium falciparum 3D7 (chloroquine-sensitive) strain, with an IC50 of 4.27±1.15 µM, while compound 1 was inactive. In silico data revealed that compound 2 showed good binding affinity to P. falciparum-Sarco endoplasmic reticulum calcium-dependent ATPase (PfATP6); however, compound 1 did not show an antiplasmodial effect due to the lack of a peroxide moiety in the chemical structure. Our data suggested that the antiplasmodial activity of compound 2 from D. esculentum might be due to the inhibition of PfATP6, which resulted in both in vitro and in silico inhibitory properties.

Effect of ergosterol peroxide derivatives on the proliferation ，migration and invasion of human triple negative breast cancer cell / 中国药房

OBJECT IVE To study the effects of ergosterol peroxide derivatives EP-3P on the proliferation ，migration and invasion of human tripe negative breast cancer cell MDA-MB- 231，and to provide reference for the development of breast cancer related drugs. METHODS MTT assay was adopted to detect the proliferation of MDA-MB- 231 cells after treated with 0（blank control），1.25，2.5，5，10，20，40 μmol/L EP-3P for 24，48 and 72 h. Wound healing assay and Transwell chamber method were adopted to detect the migration and invasion ability of MDA-MB- 231 cells after treated with 0（blank control ），5，10，20 EP-3P for 24 h. The apoptosis and cell cycle distribution were detected by flow cytometry. Western blot assay was used to detect the expressions of B-cell lympho ma-2（Bcl-2），Bcl-2 associated X protein （Bax），caspase-3，cleaved-caspase-3，cytochrome C （Cyt-C），matrix metalloproteinase- 2（MMP-2）and MMP- 9. RESULTS Compared with blank control group ，2.5，5，10，20，40 μmol/L EP-3P could significantly increase the inhibitory rate of cell proliferation （P＜0.05 or P＜0.01）in a dose and time- dependent manner. After 24 h treatment of EP- 3P（10，20 μmol/L），the rate of cell migration and the number of invasive cells were decreased significantly （P＜0.01），and cell was arrested at G 2/M stage （P＜0.05 or P＜0.01）；the apoptotic rate was increased significantly （P＜0.05）；the protein expressions of Bax ，Cyt-C and cleaved-caspase- 3 were upregulated significantly ， while those of Bcl- 2，caspase-3，MMP-2 and MMP- 9 were downregulated significantly （P＜0.01）. CONCLUSIONS EP-3P can inhibit the proliferation ，migration and invasion of human tripe negative breast cancer cells MDA-MB- 231 through mitochondrial mediated endogenous caspase pathway ，and induce the apoptosis of cells .

Antifúngicos poliénicos. Mecanismo de acción y aplicaciones / Polyenic Antifungals. Action Mechanisms and their Applications

Resumen: Los primeros compuestos con actividad antifúngica específica fueron identificados a mediados del siglo pasado como un producto del metabolismo secundario de bacterias del orden Actinomycetales, y su uso en la clínica redujo de manera importante la morbilidad y la mortalidad relacionadas con infecciones severas por hongos de varios géneros. Muchos de estos compuestos biosintéticos se caracterizan por tener una estructura química de tipo poliénico, con un número variable de dobles enlaces carbono-carbono. Actualmente, además de los fármacos poliénicos, existe otro tipo de compuestos con actividad antimicótica, como los azoles, que se utilizan con mayor frecuencia y que presentan menor toxicidad en los pacientes; sin embargo, se han documentado casos de falla terapéutica con tales compuestos, por lo que el uso de los poliénicos se ha mantenido como la mejor alternativa en esos casos. El presente trabajo brinda información acerca de las propiedades y las aplicaciones de los antifúngicos poliénicos teniendo como modelo a la anfotericina B.

Abstract The first compounds with specific antifungal activity were identified in the middle of the last century as a product of the secondary metabolism of bacteria of the order Actinomycetales, and their clinical use significantly diminished the morbidity and mortality associated with severe fungal infections. Many of such biosynthetic compounds are characterized by a chemical polygenic structure, with a variable number of carbon-carbon double bonds. Currently, besides polygenic antimycotics, there are other antifungal agents, such as the azole compounds, that have less toxicity in patients; however, cases of therapeutic failure with such compounds have been documented, therefore, the use of polygenics is still the best alternative in such cases. This review presents data about the properties and applications of antifungal-polygenic compounds using amphotericin B as a model.

Chemical constituents of alcohol extract of Mori Fructus / 中草药

Objective: To investigate the chemical constituents of alcohol extract of Mori Fructus. Methods: The chemical constituents were isolated and identified by chromatography on silica gel, Sephadex LH-20, ODS, and RP-HPLC. Their structures were elucidated on the basis of physicochemical properties and spectral analyses. Results: Twelve compounds were isolated from the alcohol extract of Mori Fructus, and identified as mullignanoside (1), (7R,8S)-4,7,9,9’-tetrahydroxy-3,3’-dimethoxy-8-O-4’-neolignan- 9’-O-β-D-glucopyranoside (2), 2-phenylethyl-β-D-glucopyranoside (3), 1’-O-phenethyl-β-D-apiofuranosyl-(1→2)-β-D-glucopyranoside (4), benzyl-O-β-D-glucopyranoside (5), ergosterol peroxide (6), (24R)-6β-hydroxy-24-ethyl-cholest-4-en-3-one (7), (22E)-5α,8α- epidioxy-24-methyl-cholesta-6,9(11),22-trien-3β-ol (8), trans-(S)-(+)-abscisic acid (9), cis-(S)-(+)-abscisic acid (10), (S)-(+)-1- methyl-abscisic-6-acid (11) and phaseic acid (12). Conclusion: Compound 1 is a new compound named mullignanoside, and compounds 2, 7-12 are isolated from this plant for the first time.

Establishment of Quality Control Method in Production of Jinshuibao Capsules / 中国实验方剂学杂志

Objective::To improve the quality control system in the production of Jinshuibao capsules, and to provide experimental basis for the follow-up research and application of this preparation. Method::High performance liquid chromatography (HPLC) was employed, the analysis was performed on a Ultimate AQ-C18 column (4.6 mm×150 mm, 5 μm). The chromatographic conditions for the determination of adenosine, guanosine and uridine were as following: mobile phase of methanol-0.1%formic acid aqueous solution for gradient elution, the flow rate of 0.4 mL·min-1, column temperature at 30 ℃, sample quantity of 10 μL, detection wavelength at 260 nm. The chromatographic conditions for the determination of ergosterol were as follows: mobile phase of methanol-water (98∶2), the flow rate of 1 mL·min-1, column temperature at 25 ℃, sample quantity of 10 μL, detection wavelength at 283 nm. Result::The main chromatographic peaks of fermented Cordyceps powder samples in different production stages showed little difference. The linear relationships of adenosine, guanosine and uridine were good (R2 >0.999), their recoveries were 106.06%, 101.25%and 105.88%, respectively. The contents of adenosine and ergosterol in 20 batches of samples extracted from 2016 to 2018 were in line with the requirements in the 2015 edition of Chinese Pharmacopoeia, the contents of guanosine and uridine were 0.97-1.36, 0.67-1.38 mg/capsule, respectively. Conclusion::The quality of Jinshuibao capsules in the market is stable. This method can be used to detect the quality of Jinshuibao capsules, and it is simple, stable and reliable.

Chemical Constituents from Polygonum cuspidatum / 中国药学杂志

OBJECTIVE: To study the chemical constituents from Polygonum cuspidatum. METHODS: The petroleum ether-ethyl acetate solvents at different ratios of 9:1, 7:1, 5:1, 3:1 and 1:1 were used as gradient eluents for silica gel column chromatography, and samples collected were further isolated and purified by preparative high performance liquid chromatography. Their structures were elucidated by physico-chemical constants and spectroscopic methods. RESULTS: Eight compounds were obtained and identified as 22(Z)-ergosterol-4,6,8,22-tetraene-3-one(1), (22E,24R)-stigma-1,4-diene-3-one(2), (E)-ethyl octadecanoic-16-enoic acid ethyl ester(3), oleanolic acid(4), emodin(5), resveratrol(6), trans-resveratrol glycoside(7) and 6'-gallic acid-4-O-D-resveratrol ester(8).CONCLUSION: Compound 1 is a new natural compound (patent No. ZL 2013 10205435.5), compound 2 and 3 are isolated from this plant for the first time.

Antifungal Mechanism of Action of Lauryl Betaine Against Skin-Associated Fungus Malassezia restricta

Betaine derivatives are considered major ingredients of shampoos and are commonly used as antistatic and viscosity-increasing agents. Several studies have also suggested that betaine derivatives can be used as antimicrobial agents. However, the antifungal activity and mechanism of action of betaine derivatives have not yet been fully understood. In this study, we investigated the antifungal activity of six betaine derivatives against Malassezia restricta, which is the most frequently isolated fungus from the human skin and is implicated in the development of dandruff. We found that, among the six betaine derivatives, lauryl betaine showed the most potent antifungal activity. The mechanism of action of lauryl betaine was studied mainly using another phylogenetically close model fungal organism, Cryptococcus neoformans, because of a lack of available genetic manipulation and functional genomics tools for M. restricta. Our genome-wide reverse genetic screening method using the C. neoformans gene deletion mutant library showed that the mutants with mutations in genes for cell membrane synthesis and integrity, particularly ergosterol synthesis, are highly sensitive to lauryl betaine. Furthermore, transcriptome changes in both C. neoformans and M. restricta cells grown in the presence of lauryl betaine were analyzed and the results indicated that the compound mainly affected cell membrane synthesis, particularly ergosterol synthesis. Overall, our data demonstrated that lauryl betaine influences ergosterol synthesis in C. neoformans and that the compound exerts a similar mechanism of action on M. restricta.

Tumor targeting and suppression effect in nude mice induced by rgd and r8 peptides modifieds liposomes loading ergosterol and cisplatin / 中国药学杂志

OBJECTIVE: To preliminarily evaluate the targeting and anti-lung cancer effect in vivo in nude mice induced by Cyclo (RGD) and R8 peptides modified ergosterol combined cisplatin liposomes. METHODS: The first step, injected RGD cyclo peptide and R8 peptide-modified, single modified or no modified ergosterol combined cisplatin liposome in the caudal vein of nude mouse bearing the tumor, the body distribution and targeting of each group under the different time points through small animals living imager were observed. The second step, continuously, dose every other day for 14 d, observating the weight of mice and the tumor growth situation. The animals were drawed blood and then were put to death, removing the tumor, the spleen and the lung tissue of all the mice. As the index of the tumor weight, the tumor suppression effect, the level of TGF-β1, TIMPs and TNF-α in serum, the spleen index and changes of the tumor and lung tissue, investigate the tumor suppression effect in mice of the liposomes preliminary. RESULTS: The targeting result of tumor-bearing nude mice displays that the fluorescence intensity of RGD and R8 peptides-modified liposome is the highest and the targeting is most obvious under high concentration and other group of liposome are weaker. Preliminary pharmacodynamics results show that each dosage group of mice have no obvious change in body weight and the high and middle dose group of RGD and R8 peptides-modified liposomes has tumor suppression effect obviously. The high dose group of RGD and R8 peptides-modified liposome is the most significant. It has high expression of cytokines (TNF-α) in serum. The spleen index of middle and low dose group of RGD and R8 peptides-modified liposomes significantly increased compared with positive medicine group. CONCLUSION: RGD cyclo peptide and R8 peptide-modified ergosterol combined cisplatin targeting liposome drug delivery system further improves the tumor targeting and anti-lung cancer effect in vivo.

Chemical constituents from Rabdosia phyllopoda / 中草药

Objective: To investigate the chemical constituents of Rabdosia phyllopoda. Methods: The chemical constituents were isolated and purified by chromatography with silica gel, Sephadex LH-20 and by semi-preparative HPLC, and their structures were identified by analysis of spectroscopic evidences and physicochemical properties. Results: A total of 14 constituents were isolated from R. phyllopoda and elucidated as (Z)-1,1’-biindenyliden (1), β,β-carotene (2), germacrene B (3), 2α,3α,24-trihydrxyolean-12- en-28 oic acid (4), ferulic acid hexacosanyl ester (5), ergosterol peroxide (6), ursolic acid (7), α-amyrin acetate (8), obtusalin (9), betulinic acid (10), 2α,3α,19α-trihydroxy-12-en-28-urs acid (11), 2α,3α-dihydroxyl-12-ene-28 oic acid (12), 2α-hydroxyursolic acid (13), and lutein (14). Conclusion: The 14 compounds are isolated from this plant for first time, and compound 1, 3, 6, 9, 14 are isolated from the plants of R. phyllopoda for the first time.

Sesquiterpenes and steroids from Subergorgia suberosa / 中草药

Objective: To investigate the chemical constitutes of Subergorgia suberosa from South China Sea. Methods: We carried out the chemical separation of EtOH extract of S. suberosa based on the chromatographic techniques such as medium pressure preparative chromatography, silica, sephadex LH-20, ODS column chromatography, and preparative thin layer chromatography (PTLC). The NMR and MS spectroscopic techniques, together with comparison with literature data, were used for structural elucidation of compounds. Results: Fourteen compounds including sesquiterpenes and steroids were isolated from S. suberosa. These compounds were identified as subergorgic acid methyl ester (1), 2β-hydroxy subergorgic acid methyl ester (2), subergorgic acid (3), suberosenol A (4), suberosenol B (5), suberosenone (6), suberosanone (7), 3β-hydroxy-5β-pregnan-20-one (8), 3β-hydroxy-5α-pregnan-20-one (9), 3α-hydroxy-5β-pregnan-20-one (10), pregnan-4-ene-3,20-dione (11), 3β-hydroxy-pregnan-5-ene-20-one (12), cholesterol (13), and 5α,8α-epidioxyergosta-6,22-dien-3β-ol (14), respectively. Conclusion: Compounds 5 and 10 are isolated from the family Subergorgiidae for the first time. Compound 14 is isolated from S. suberosa for the first time.

Determination of Ergosterol as a Potential Biomarker in Pathogenic Medically Important Fungal Isolates / Jurnal Sains Kesihatan Malaysia

Ergosterol, a component of fungal cell membrane, has been frequently detected as an indicator of fungal presence and massin environmental samples like soil. However, its detection in major pathogenic fungal species has not been investigated.In this study, the ergosterol contents of ten pathogenic fungal species were determined. Liquid chromatography was usedfor the detection and quantification of ergosterol extracted from fungal broth cultures. Results showed that ergosteroleluted as a single, well resolved peak in the chromatogram profiles of all tested fungi. Based upon relative amounts ofergosterol produced per fungal mycelial dry weight, three groups of fungal pathogens were identified, namely low ergosterol(Aspergillus niger, Candida albicans and Cryptococcus neoformans at 4.62, 6.29 and 7.08 μg/mg, respectively), mediumergosterol (Fusarium solani, Aspergillus fumigatus, Mucor sp., Penicillium sp., Cryptococcus gattii and Rhizopus sp.at 9.40, 10.79, 10.82, 11.38, 12.60 and 13.40 μg/mg, respectively), and high ergosterol (Candida tropicalis at 22.84 μg/mg), producers. Ergosterol was not detectable in bacterial samples, which were included as controls. This first report onergosterol detection in major pathogenic fungal species indicates that ergosterol may be used as a biomarker to diagnoseinvasive fungal infections in clinical sampl

Butyl alcohol extract of Baitouweng decoction inhibits Candida albicans cell membrane / 中国中药杂志

To study the inhibitory effect of butyl alcohol extract of Baitouweng decoction(BAEB) on Candida albicans cell membrane. The effects of BAEB on the activity of C. albicans were observed by Spot assay. The changes of intracellular osmotic pressure of C. albicans after BAEB intervention were detected by microtiter plate reader. The effect of BAEB on cell membrane permeability of C. albicans were observed by fluorescence microscopy. The content of ergosterol in C. albicans cell membrane was detected by high performance liquid chromatography, and the expression of ergosterol biosynthesis related genes in cell membrane was detected by qRT-PCR. The results showed that the activity of C. albicans was significantly decreased in 256, 512 and 1 024 mg•L⁻¹ BAEB group. The intracellular glycerol content of C. albicans was significantly increased in 512 and 1 024 mg•L⁻¹ BAEB group(P<0.05). The gene HOG1 associated with intracellular osmotic pressure of C. albicans was down-regulated by 9.1, 9.3 and 5.5 times, respectively. C. albicans with red fluorescent were increased significantly in 512 and 1 024 mg•L⁻¹ BAEB group. The peak area of ergosterol in the 1 024 mg•L⁻¹ BAEB group was 35.884 95, with a significant difference(P<0.05); ERG1, ERG2, ERG3, ERG4, ERG5, ERG6, ERG10, ERG11, ERG13, ERG24, ERG25, ERG251, ERG26 and UPC2 were down-regulated by 6.58, 4.89, 4.15, 9.24,3.41, 9.84, 3.08, 7.50, 5.53, 5.90, 2.45, 3.25,1.98 and 10.07 times respectively in 1 024 mg•L⁻¹ BAEB group. The study indicated that BAEB could inhibit ergosterol and its biosynthesis related genes expression in the cell membrane and inhibit the activity of C. albicans.

Anti-platelet Aggregation Activity of Ergosterol in Vitro / 医药导报

Objective To evaluate the anti-platelet aggregation effect of ergosterol in vitro and explore the preliminary mechanism. Methods The anti-platelet aggregation activity of ergosterol was assessed in vitro on rabbit platelet aggregation. Different inducers, ADP ( 4 μmol?L-1 ) , collagen ( 4 μg?mL-1 ) , arachidonic acid ( AA, 1 mmol?L-1 ) and thrombin (0.5 U?mL-1), and blockers (ozagrel, dipyridamole, clopidogrel and aspirin) were applied to observe the potential targets of ergosterol, platelet aggregation induced by ADP (0, 1, 2, 4, 6μmol?L-1) or fibrinogen (0, 1, 2, 4, 6, 10 mg?mL-1). Results Ergosterol exhibited an obvious anti-platelet aggregation effect in vitro with IC50 values on different inducers ( ADP, collagen, AA and thrombin) of (19.3±0.8), (23.4±1.2), (26.7±0.7), (32.9±1.5) μmol?L-1, respectively. Conclusion Ergosterol can significantly inhibit aggregation and activation of platelet. It provides experimental basis for full exploration of ergosterol and development of novel anti-platelet aggregation drugs.

Antifungal Activity of Aspidin BB from Dryopteris fragrans against Trichophyton rubrum Involved Inhibition of Ergosterol Biosynthesis / 中草药·英文版

Objectives Aspidin BB, a typical phloroglucinol derivative from Dryopteris fragrans, possesses significant antifungal property. This study aimed to investigate potential mechanism of antifungal activity of Aspidin BB against Trichophyton rubrum which is the most common pathogens responsible for chronic dermatophytosis. Methods The minimum inhibitory concentration (MIC) of Aspidin BB against strains was determined by broth microdilution. The effects of Aspidin BB on ergosterol biosynthesis were investigated by content determination based on UPLC method. Besides, the effects of drugs on squalene epoxidase (SE) in T. rubrum cell membrane were analyzed. Results MIC value of Aspidin BB against T. rubrum was 25.0 μg/mL. Aspidin BB reduced ergosterol content significantly, but no notable effect on squalene epoxidase activity. Conclusion The results suggested that Aspidin BB inhibited ergosterol biosynthesis. However, it was not squalene epoxidase but other components may sever as possible targets in ergosterol biosynthesis pathway.

Effects of three active components in Cordyceps sinensis regulating liver endothelial cells functions based on angiogenesis / 中草药

Objective: To investigate the effects of three active components in Cordyceps sinensis (cordycepin, adenosine, and ergosterol) on angiogenesis and the function of hepatic endothelial cells. Methods: Chick chorioallantoic membrane was used to record angiogenesis area, and transgenic zebrafish was used to evaluate the changes of intersegmental vascular and alkaline phosphatase. The toxicity of three active components was assayed in SK-HEP-1 cells by MTT. The proliferation of SK-HEP-1 cells was induced by vascular endothelial growth factor (VEGF), with sorafenib as the positive control. Cell proliferation was analyzed by MTT assay. Cell migration and tube formation were observed by Matrigel and Transwell assay, respectively. Fluorescence probe method was used to detect the levels of intracellular nitric oxide (NO) and nitric oxide synthase (NOS). Results: Adenosine and ergosterol inhibited angiogenesis of chick chorioallantoic membrane, and decreased the number of transgenic zebrafish intersegmental vascular. However, cordycepin can promote angiogenesis of chick chorioallantoic membrane. Compared with the blank control group, VEGF induced SK-HEP-1 cells proliferation, promoted cells migration and tube formation, further significantly increased the levels of NO and NOS in SK-HEP-1. All three active components inhibited the proliferation, tube formation of SK-HEP-1 and decreased the levels of NO and NOS in a dose-dependent manner. Moreover, adenosine and ergosterol inhibited SK-HEP-1 cells migration significantly. Conclusion: Three active components, especially adenosine, could inhibit SK-HEP-1 cells proliferation, migration and tube formation in different degrees. And the mechanism is related to the inhibition of intracellular NO levels and NOS activity.

Ergosterol isolated from the basidiomycete Pleurotus salmoneostramineus affects Trypanosoma cruzi plasma membrane and mitochondria

Abstract Background Major drawbacks of the available treatment against Chagas disease (American trypanosomiasis) include its toxicity and therapeutic inefficiency in the chronic phase of the infection, which makes it a concern among neglected diseases. Therefore, the discovery of alternative drugs for treating chronic Chagas disease requires immediate action. In this work, we evaluated the mushroom Pleurotus salmoneostramineus in the search for potential antiparasitic compounds. Methods Fruit bodies of the basidiomycete Pleurotus salmoneostramineus were triturated and submitted to organic solvent extraction. After liquid-liquid partition of the crude extract, three fractions were obtained and the bioguided fractionation study was conducted to isolate the active metabolites. The elucidation of the chemical structure was performed using GC-MS and NMR techniques. The biological assays for antiparasitic activity were carried out using trypomastigotes of Trypanosoma cruzi and murine macrophages for mammalian cytotoxicity. The mechanism of action of the isolated compound used different fluorescent probes to evaluate the plasma membrane permeability, the potential of the mitochondrial membrane and the intracellular levels of reactive oxygen species (ROS). Results The most abundant fraction showing the antiparasitic activity was isolated and chemically elucidated, confirming the presence of ergosterol. It showed anti-Trypanosoma cruzi activity against trypomastigotes, with an IC50 value of 51.3 g/mL. The compound demonstrated no cytotoxicity against mammalian cells to the maximal tested concentration of 200 g/mL. The mechanism of action of ergosterol in Trypanosoma cruzi trypomastigotes resulted in permeabilization of the plasma membrane, as well as depolarization of mitochondrial membrane potential, leading to parasite death. Nevertheless, no increase in ROS levels could be observed, suggesting damages to plasma membrane rather than an induction of oxidative stress in the parasite. Conclusions The selection of naturally antiparasitic secondary metabolites in basidiomycetes, such as ergosterol, may provide potential scaffolds for drug design studies against neglected diseases.

Ergosterol isolated from the basidiomycete Pleurotus salmoneostramineus affects Trypanosoma cruzi plasma membrane and mitochondria

Background Major drawbacks of the available treatment against Chagas disease (American trypanosomiasis) include its toxicity and therapeutic inefficiency in the chronic phase of the infection, which makes it a concern among neglected diseases. Therefore, the discovery of alternative drugs for treating chronic Chagas disease requires immediate action. In this work, we evaluated the mushroom Pleurotus salmoneostramineus in the search for potential antiparasitic compounds. Methods Fruit bodies of the basidiomycete Pleurotus salmoneostramineus were triturated and submitted to organic solvent extraction. After liquid-liquid partition of the crude extract, three fractions were obtained and the bioguided fractionation study was conducted to isolate the active metabolites. The elucidation of the chemical structure was performed using GC-MS and NMR techniques. The biological assays for antiparasitic activity were carried out using trypomastigotes of Trypanosoma cruzi and murine macrophages for mammalian cytotoxicity. The mechanism of action of the isolated compound used different fluorescent probes to evaluate the plasma membrane permeability, the potential of the mitochondrial membrane and the intracellular levels of reactive oxygen species (ROS). Results The most abundant fraction showing the antiparasitic activity was isolated and chemically elucidated, confirming the presence of ergosterol. It showed anti-Trypanosoma cruzi activity against trypomastigotes, with an IC50 value of 51.3 μg/mL. The compound demonstrated no cytotoxicity against mammalian cells to the maximal tested concentration of 200 μg/mL. The mechanism of action of ergosterol in Trypanosoma cruzi trypomastigotes resulted in permeabilization of the plasma membrane, as well as depolarization of mitochondrial membrane potential, leading to parasite death. Nevertheless, no increase in ROS levels could be observed, suggesting damages to plasma membrane rather than an induction of oxidative stress in the parasite. Conclusions The selection of naturally antiparasitic secondary metabolites in basidiomycetes, such as ergosterol, may provide potential scaffolds for drug design studies against neglected diseases.(AU)