scholarly journals Ellagic Acid Inhibits Trichophyton rubrum Growth via Affecting Ergosterol Biosynthesis and Apoptotic Induction

2020 ◽  
Vol 2020 ◽  
pp. 1-8
Author(s):  
Zhi-Jian Li ◽  
Amima Abula ◽  
Abudumijiti Abulizi ◽  
Chun Wang ◽  
Qin Dou ◽  
...  
Keyword(s):  
Cell Membrane ◽  
Ellagic Acid ◽  
Cell Apoptosis ◽  
Trichophyton Rubrum ◽  
Ergosterol Biosynthesis ◽  
Dependent Manner ◽  
Biosynthesis Pathway ◽  
Gene Expressions ◽  
Fungal Cell

Background. Trichophyton rubrum, among other dermatophytes, is a major causative agent for superficial dermatomycoses like onychomycosis and tinea pedis, especially among pediatric and geriatric populations. Ellagic acid (EA) and shikonin (SK) have been reported to have many bioactivities, including antifungal activity. However, the mechanism of EA and SK on Trichophyton rubrum has not yet been reported. Objectives. The purposes of this study were to evaluate the antifungal activities of EA and SK against Trichophyton rubrum and to illuminate the underlying action mechanisms. Methods. The effect of EA (64, 128, and 256 μg/mL) and SK (8, 4, and 2 μg/mL) on Trichophyton rubrum was investigated with different doses via detecting cell viability, ultrastructure with using a scanning electron microscope (SEM), cell apoptosis and necrosis by using the flow cytometry instrument technique (FCIT), and the ergosterol biosynthesis pathway-related fungal cell membrane key gene expressions in vitro. Results. SEM detection revealed that the T. rubrum cell surface was shrivelled, folded, and showed deformation and expansion, visible surface peeling, and broken hyphae, and cell contents overflowed after being treated with EA and SK; the cell apoptosis rate was significantly increased in dose-dependent manner after T. rubrum was treated with EA and SK; the qPCR results showed that mRNA expression of MEP4 and SUB1 was downregulated in EA- and SK-treated groups. Conclusions. Overall, our results revealed the underlying antifungal mechanism of EA and SK, which may be related to the destruction of the fungal cell membrane and inhibition of C14 demethylase and the catalytic rate of squalene cyclooxidase in the ergosterol biosynthesis pathway via downregulation of MEP4 and SUB1, suggesting that EA and SK have the potential to be developed further as a natural antifungal agent for clinical use.

scholarly journals NBM-HD-1: A Novel Histone Deacetylase Inhibitor with Anticancer Activity

2012 ◽  
Vol 2012 ◽  
pp. 1-11 ◽  
Author(s):  
Wei-Jan Huang ◽  
Yu-Chih Liang ◽  
Shuang-En Chuang ◽  
Li-Ling Chi ◽  
Chi-Yun Lee ◽  
...  
Keyword(s):  
Anticancer Activity ◽  
Cancer Cells ◽  
Anticancer Agents ◽  
Xenograft Model ◽  
Cyclin B1 ◽  
Dependent Manner ◽  
Cell Cycle Regulators ◽  
Anticancer Activities ◽  
Gene Expressions

HDAC inhibitors (HDACis) have been developed as promising anticancer agents in recent years. In this study, we synthesized and characterized a novel HDACi, termed NBM-HD-1. This agent was derived from the semisynthesis of propolin G, isolated from Taiwanese green propolis (TGP), and was shown to be a potent suppressor of tumor cell growth in human breast cancer cells (MCF-7 and MDA-MB-231) and rat glioma cells (C6), with an IC50ranging from 8.5 to 10.3 μM. Western blot demonstrated that levels of p21(Waf1/Cip1), gelsolin, Ac-histone 4, and Ac-tubulin markedly increased after treatment of cancer cells with NBM-HD-1. After NBM-HD-1 treatment for 1–4 h, p-PTEN and p-AKT levels were markedly decreased. Furthermore, we also found the anticancer activities of NBM-HD-1 in regulating cell cycle regulators. Treatment with NBM-HD-1,p21(Waf1/Cip1)gene expression had markedly increased whilecyclin B1andD1gene expressions had markedly decreased. On the other hand, we found that NBM-HD-1 increased the expressions of tumor-suppressor genep53in a dose-dependent manner. Finally, we showed that NBM-HD-1 exhibited potent antitumor activity in a xenograft model. In conclusion, this study demonstrated that this compound, NBM-HD-1, is a novel and potent HDACi with anticancer activityin vitroandin vivo.

scholarly journals Synthesis and In Vitro Antitumor Activity of Novel Bivalent β-Carboline-3-carboxylic Acid Derivatives with DNA as a Potential Target

2018 ◽  
Vol 19 (10) ◽  
pp. 3179 ◽  
Author(s):  
Hongling Gu ◽  
Na Li ◽  
Jiangkun Dai ◽  
Yaxi Xi ◽  
Shijun Wang ◽  
...  
Keyword(s):  
Antitumor Activity ◽  
Cell Apoptosis ◽  
Docking Studies ◽  
In Vitro Cytotoxicity ◽  
Dependent Manner ◽  
Cycle Arrest ◽  
Dna Binding Affinity ◽  
Dose Dependent ◽  
Mcf 7

A series of novel bivalent β-carboline derivatives were designed and synthesized, and in vitro cytotoxicity, cell apoptosis, and DNA-binding affinity were evaluated. The cytotoxic results demonstrated that most bivalent β-carboline derivatives exhibited stronger cytotoxicity than the corresponding monomer against the five selected tumor cell lines (A549, SGC-7901, Hela, SMMC-7721, and MCF-7), indicating that the dimerization at the C3 position could enhance the antitumor activity of β-carbolines. Among the derivatives tested, 4B, 6i, 4D, and 6u displayed considerable cytotoxicity against A549 cell line. Furthermore, 4B, 6i, 4D, and 6u induced cell apoptosis in a dose-dependent manner, and caused cell cycle arrest at the S and G2/M phases. Moreover, the levels of cytochrome C in mitochondria, and the expressions of bcl-2 protein, decreased after treatment with β-carbolines, which indicated that 6i and 6u could induce mitochondria-mediated apoptosis. In addition, the results of UV-visible spectral, thermal denaturation, and molecular docking studies revealed that 4B, 6i, 4D, and 6u could bind to DNA mainly by intercalation.

Inhibitory activity of isoniazid and ethionamide against Cryptococcus biofilms

2015 ◽  
Vol 61 (11) ◽  
pp. 827-836 ◽  
Author(s):  
Rossana de Aguiar Cordeiro ◽  
Rosana Serpa ◽  
Francisca Jakelyne de Farias Marques ◽  
Charlline Vládia Silva de Melo ◽  
Antonio José de Jesus Evangelista ◽  
...  
Keyword(s):  
Cell Membrane ◽  
Biofilm Formation ◽  
Inhibitory Effect ◽  
Cell Membrane Permeability ◽  
Planktonic Cells ◽  
Fungal Cell ◽  
Antituberculosis Drugs ◽  
Opportunistic Mycoses ◽  
Cryptococcus Spp

In recent years, the search for drugs to treat systemic and opportunistic mycoses has attracted great interest from the scientific community. This study evaluated the in vitro inhibitory effect of the antituberculosis drugs isoniazid and ethionamide alone and combined with itraconazole and fluconazole against biofilms of Cryptococcus neoformans and Cryptococcus gattii. Antimicrobials were tested at defined concentrations after susceptibility assays with Cryptococcus planktonic cells. In addition, we investigated the synergistic interaction of antituberculosis drugs and azole derivatives against Cryptococcus planktonic cells, as well as the influence of isoniazid and ethionamide on ergosterol content and cell membrane permeability. Isoniazid and ethionamide inhibited both biofilm formation and viability of mature biofilms. Combinations formed by antituberculosis drugs and azoles proved synergic against both planktonic and sessile cells, showing an ability to reduce Cryptococcus biofilms by approximately 50%. Furthermore, isoniazid and ethionamide reduced the content of ergosterol in Cryptococcus spp. planktonic cells and destabilized or permeabilized the fungal cell membrane, leading to leakage of macromolecules. Owing to the paucity of drugs able to inhibit Cryptococcus biofilms, we believe that the results presented here might be of interest in the designing of new antifungal compounds.

scholarly journals Toxicity Effects of Methylene Blue on Rat Intervertebral Disc Annulus Fibrosus Cells

2019 ◽  
Vol 2 (22.2) ◽  
pp. 155-164
Author(s):  
Liang Zhang
Keyword(s):  
Methylene Blue ◽  
Cell Apoptosis ◽  
Annulus Fibrosus ◽  
Caspase 3 ◽  
In Vitro Study ◽  
Collagen Type I ◽  
Type I ◽  
Dependent Manner ◽  
Concentration Dependent Manner

Background: There is an increasing local application of methylene blue (MB) in the treatment of discogenic low back pain (LBP) and percutaneous transforaminal endoscopic discectomy (PTED) procedures. MB could generate DNA damage and induce apoptosis in different cell types; however, the effects of MB on intervertebral disc (IVD) annulus fibrosus (AF) cells are not clearly understood. Objective: The objective of this study was to investigate the effects of different concentrations of MB on rat AF cells in vitro. Study Design: This study used an experimental design. Setting: This research was conducted at the Orthopaedic Institute of the Clinical Medical College of Yangzhou University. Methods: AF cells were isolated and cultured with different concentrations of MB (0, 2, 20, and 200 μg/mL) and assessed to determine the possible cytotoxic effects of MB. The cell proliferation was detected by Cell Counting Kit-8 (CCK-8) assay. The inverted phase-contrast microscopy was used to perform morphological observation of apoptotic cells, and flow cytometry was used to measure the incidence of cell apoptosis. The mRNA and protein expression levels of apoptosis-associated genes (caspase-3, Bcl-2, and Bax) and other related genes (collagen type I, transforming growth factor β1 [TGF-β1], fibroblast growth factor [bFGF], and tissue inhibitor of metalloproteinase-1 [TIMP-1]) were analyzed by quantitative real-time PCR (RT-PCR) and Western blotting. Results: Our results indicated that MB reduced cell viability in a concentration- and timedependent manner. MB also induced marked AF cell apoptosis in a concentration-dependent manner observed by inverted phase-contrast microscopy, flow cytometry, and indicated by the increased expression of caspase-3. Both RT-PCR and Western blotting revealed significant upregulation of Bax and caspase-3 expression levels accompanied by decreased expression of Bcl2 in a concentration-dependent manner. Moreover, collagen type I, TGF-β1, bFGF, and TIMP-1 mRNA and protein levels were also found to be decreased by MB in a concentration-dependent manner. Limitations: Limitations of this study were the in vitro study design and lack of in vivo validation of the observed effects of MB on human IVD cells. Conclusions: Our results indicate that a high concentration of MB can not only inhibit proliferation and paracrine function of AF cells, but can also induce cell apoptosis in a concentration-dependent manner, suggesting that it is necessary to choose low concentrations of MB in practical application and limit the use of MB in the treatment of discogenic LBP to research protocols. Key words: Methylene blue, annulus fibrosus cell, proliferation, apoptosis, paracrine

scholarly journals SAT-124 Optimization of Experimental Conditions for Mimicking Hypoxia in Cultured Breast Cancer Cells by Using Cobalt(II) Chloride (CoCl2)

2020 ◽  
Vol 4 (Supplement_1) ◽  
Author(s):  
Karin Chen ◽  
Leo Satlof ◽  
Udithi Kothapalli ◽  
Noah Ziluck ◽  
Maribel Lema ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cancer Cells ◽  
Cell Apoptosis ◽  
Breast Cancer Cells ◽  
Nuclear Translocation ◽  
Breast Cancer Cell Lines ◽  
Mrna Levels ◽  
Dependent Manner ◽  
Experimental Conditions

Abstract Hypoxia is a common phenomenon in solid tumor development caused by a decrease in either oxygen concentration or oxygen pressure as a result of rapid tumor cell growth. Hypoxia is characterized by stabilization of the alpha subunit of the hypoxia-inducible factor (HIF-1α) and its nuclear translocation and heterodimerization with HIF-1β. Activation of this signaling pathway involves multiple downstream effectors including carbonic anhydrase 9 (CA9, s. CAIX). A reliable method to mimic hypoxia utilizes cobalt(II) chloride (CoCl2), which directly induces the expression of HIF-1α. The aim of this study was to optimize the experimental conditions for CoCl2 treatment of breast cancer cells in vitro using three human breast cancer cell lines (MDA-MB-231, T-47D, and MCF-7 cells). We performed time- and concentration-response experiments, using various concentrations of CoCl2 (50, 100, 200, and 300 μM) for 24 and 48 hours, and measured the expression of HIF-1α and CA9 by qRT-PCR and Western blot analyses. Results demonstrated that CoCl2 downregulated HIF-1α mRNA levels but upregulated CA9 mRNA levels in a concentration- and time-dependent manner. Concomitantly, CoCl2 treatment resulted in a significant induction of HIF-1α protein levels. We further investigated the effect of the CoCl2 concentrations listed above on cell apoptosis using an in situ apoptosis detection kit. The results demonstrated that concentrations of CoCl2 up to 100 μM had no significant effect on cell apoptosis.

scholarly journals Artemether Exhibits Amoebicidal Activity against Acanthamoeba castellanii through Inhibition of the Serine Biosynthesis Pathway

2015 ◽  
Vol 59 (8) ◽  
pp. 4680-4688 ◽  
Author(s):  
Yihong Deng ◽  
Wei Ran ◽  
Suqin Man ◽  
Xueping Li ◽  
Hongjian Gao ◽  
...  
Keyword(s):  
Acanthamoeba Castellanii ◽  
Dependent Manner ◽  
Biosynthesis Pathway ◽  
Content Type ◽  
Phosphoserine Aminotransferase ◽  
Proteomic Approach ◽  
Phosphoglycerate Dehydrogenase ◽  
Serine Biosynthesis ◽  
Amoebicidal Activity

ABSTRACTAcanthamoebasp. parasites are the causative agents ofAcanthamoebakeratitis, fatal granulomatous amoebic encephalitis, and cutaneous infections. However, there are currently no effective drugs for these organisms. Here, we evaluated the activity of the antimalarial agent artemether againstAcanthamoeba castellaniitrophozoites and identified potential targets of this agent through a proteomic approach. Artemether exhibitedin vitroamoebicidal activity in a time- and dose-dependent manner and induced ultrastructural modification and cell apoptosis. The iTRAQ quantitative proteomic analysis identified 707 proteins that were differentially expressed after artemether treatment. We focused on phosphoglycerate dehydrogenase and phosphoserine aminotransferase in the serine biosynthesis pathway because of their importance to the growth and proliferation of protozoan and cancer cells. The expression of these proteins inAcanthamoebawas validated using quantitative real-time PCR and Western blotting after artemether treatment. The changes in the expression levels of phosphoserine aminotransferase were consistent with those of phosphoglycerate dehydrogenase. Therefore, the downregulation of phosphoserine aminotransferase may be due to the downregulation of phosphoglycerate dehydrogenase. Furthermore, exogenous serine might antagonize the activity of artemether againstAcanthamoebatrophozoites. These results indicate that the serine biosynthesis pathway is important to amoeba survival and that targeting these enzymes would improve the treatment ofAcanthamoebainfections. Artemether may be used as a phosphoglycerate dehydrogenase inhibitor to control or blockAcanthamoebainfections.

In vitro Antifungal Activity and Mechanism of Action of Tea Polyphenols and Tea Saponin against Rhizopus stolonifer

2015 ◽  
Vol 25 (4) ◽  
pp. 269-276 ◽  
Author(s):  
Xiaodong Jiang ◽  
Kejue Feng ◽  
Xiaoping Yang
Keyword(s):  
Cell Membrane ◽  
Oxidative Damage ◽  
Mechanism Of Action ◽  
Soluble Sugar ◽  
Tea Polyphenols ◽  
Cell Membrane Permeability ◽  
Dependent Manner ◽  
Rhizopus Stolonifer ◽  
Tea Saponin

The in vitro antifungal activities and mechanism of action of tea polyphenols (TP), tea saponin (TS) and their combination were evaluated against <i>Rhizopus stolonifer</i>. The results showed that both TP and TS inhibited the mycelial growth in a dose-dependent manner, and their combination at the ratio of 7:3 exhibited synergistic antifungal interaction. We also observed that the treatment of TP or TS significantly induced the production of H<sub>2</sub>O<sub>2</sub> and resulted in membrane lipid peroxidation, thus leading to an increase in cell membrane permeability and the leakage of K<sup>+</sup>, soluble protein and soluble sugar. Moreover, combining them for treatment increased the induction of H<sub>2</sub>O<sub>2</sub> production and oxidative damage. Scanning electron microscopic observations also showed the damage to the hyphal cell structure. It was concluded that TP, TS and their combination inhibit the growth of <i>R. stolonifer</i> through the induction of H<sub>2</sub>O<sub>2</sub> production, leading to cell membrane oxidative damage and intracellular constituent leakage. These findings suggest that TP and TS can potentially be used as an alternative to control postharvest fruit diseases caused by <i>R. stolonifer.</i>

In vitro leishmanicidal activity of antimicrobial peptide KDEL against Leishmania tarentolae

2019 ◽  
Vol 51 (12) ◽  
pp. 1286-1292 ◽  
Author(s):  
Lili Cao ◽  
Weina Jiang ◽  
Songgao Cao ◽  
Panpan Zhao ◽  
Juan Liu ◽  
...  
Keyword(s):  
Cell Apoptosis ◽  
Therapeutic Potential ◽  
Surface Membrane ◽  
Membrane Integrity ◽  
Annexin V ◽  
Protozoan Parasite ◽  
Immune Defense ◽  
Dependent Manner ◽  
Leishmania Tarentolae

Abstract Leishmaniasis, caused by the intracellular protozoan parasite Leishmania, remains an important neglected tropical infectious disease. Infection may be lethal if untreated. Currently, the available drugs for the disease are limited by high toxicity and drug resistance. There is an urgent need to develop novel anti-leishmanial strategies. Antimicrobial peptides (AMPs) have been described as the first-line immune defense against pathogenic microbes and are being developed as emerging anti-parasitic therapies. In the present study, we showed the anti-leishmanial activity of the synthetic 4-amino acid peptide lysine, aspartic acid, glutamic acid, and leucine (KDEL), the endoplasmic reticulum retention sequence, against Leishmania tarentolae promastigote and amastigote. Different concentrations of KDEL peptides were incubated with promastigotes, MTT viability assay, and promastigote assay were carried out. Macrophages infected with GFP-transfected L. tarentolae promastigotes were incubated with KDEL peptides, and the anti-amastigote activity of the KDEL peptides was measured by fluorescence microscopy. The damage of L. tarentolae was observed by light microscopy and electron microscopy. The cell apoptosis was analyzed using the Annexin V-FITC/PI apoptosis detection kit and mitochondrial membrane potential assay kit and by flow cytometry. Results showed that L. tarentolae was susceptible to KDEL peptides in a dose-dependent manner, and KDEL peptides disrupted the surface membrane integrity and caused cell apoptosis. In our study, we found for the first time an AMP KDEL from Pseudomonas aeruginosa and proved its significant therapeutic potential as a novel anti-leishmanial drug.

scholarly journals Hepatotoxicity of Pyrrolizidine Alkaloid Compound Intermedine: Comparison with Other Pyrrolizidine Alkaloids and Its Toxicological Mechanism

Toxins ◽  
2021 ◽  
Vol 13 (12) ◽  
pp. 849
Author(s):  
Ziqi Wang ◽  
Haolei Han ◽  
Chen Wang ◽  
Qinqin Zheng ◽  
Hongping Chen ◽  
...  
Keyword(s):  
Cell Apoptosis ◽  
Pyrrolizidine Alkaloids ◽  
Pyrrolizidine Alkaloid ◽  
Herbal Medicines ◽  
Cell Counting ◽  
Systematic Evaluation ◽  
Sinusoidal Obstruction Syndrome ◽  
Dependent Manner ◽  
Primary Mouse

Pyrrolizidine alkaloids (PAs) are common secondary plant compounds with hepatotoxicity. The consumption of herbal medicines and herbal teas containing PAs is one of the main causes of hepatic sinusoidal obstruction syndrome (HSOS), a potentially life-threatening condition. The present study aimed to reveal the mechanism underlying the cytotoxicity of intermedine (Im), the main PA in Comfrey. We evaluated the toxicity of the retronecine-type PAs with different structures to cell lines derived from mammalian tissues, including primary mouse hepatocytes, human hepatocytes (HepD), mouse hepatoma-22 (H22) and human hepatocellular carcinoma (HepG2) cells. The cytotoxicity of Im to hepatocyte was evaluated by using cell counting kit-8 assay, colony formation experiment, wound healing assay and dead/live fluorescence imaging. In vitro characterization showed that these PAs were cytotoxic and induced cell apoptosis in a dose-dependent manner. We also demonstrated that Im induced cell apoptosis by generating excessive reactive oxygen species (ROS), changing the mitochondrial membrane potential and releasing cytochrome c (Cyt c) before activating the caspase-3 pathway. Importantly, we directly observed the destruction of the cell mitochondrial structure after Im treatment through transmission electron microscopy (TEM). This study provided the first direct evidence of Im inducing hepatotoxicity through mitochondria-mediated apoptosis. These results supplemented the basic toxicity data of PAs and facilitated the comprehensive and systematic evaluation of the toxicity caused by PA compounds.

Single-Agent Bruton's Tyrosine Kinase (Btk) Inhibitor and Single-Agent Carfilzomib Induce Cell Apoptosis, Arrest Cell Growth and Downregulate NF-Kb Activity In Mantle Cell Lymphoma

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 4971-4971 ◽  
Author(s):  
Zhishuo Ou ◽  
Lan V Pham ◽  
Liang Zhang ◽  
Archito T. Tamayo ◽  
John Lee ◽  
...  
Keyword(s):  
Cell Growth ◽  
B Cell ◽  
Cell Apoptosis ◽  
Cell Lymphoma ◽  
Single Agent ◽  
Therapeutic Agents ◽  
Dependent Manner ◽  
Mantle Cell ◽  
Btk Inhibitor

Abstract Abstract 4971 Mantle cell lymphoma (MCL) is an aggressive histotype of B-cell non-Hodgkin lymphoma with poor prognoses. Discovery of less-toxic and better therapeutic agents is an ongoing challenge. Bruton tyrosine kinase (BTK), is identified as an essential kinase for B-cell survival, activated through the B-cell receptor (BCR) pathway. Recent studies have indicated that BTK mediates NF-kB activation. The potential therapeutic agent PCI-32765, a novel BTK inhibitor (Pharmacyclics, Sunnyvale, CA) have shown effective preliminary targeting of these pathways in MCL. Carfilzomib, a 2nd generation proteasome inhibitor (Onyx Pharm. Inc., Emeryville, CA) proven effective in myeloma in clinical trial is also effective in MCL in vitro and in vivo in pre-clinical studies. The objective of our study was to evaluate the therapeutic efficacy of PCI-32765 and carfilzomib in MCL, and elucidate the mechanisms of their actions. We initially screened 12 MCL cell lines and discovered that Btk is constitutively phosphorylated in all MCL cells. Representative MCL cell lines, both classic and blastoid variant, were utilized for in vitro studies. PCI-32765 effectively inhibited phospho-BTK, leading to reduced MCL cell growth with IC:50 values range from 10–50 uM. PCI-32765 also induced MCL cell apoptosis in both dose- and time-dependent manner as well as carfilzomib. Using luciferase pGL3 reporter plasmid (6xNF-kB-CD40L/TKm), we demonstrated that both PCI-32765 and carfilzomib down-regulated the reporter with a dosage dependent manner. Our data suggest that strategic targeting of growth/survival pathways with novel therapeutic agents PCI-32765 and carfilzomib should provide a novel therapy regimen for patients with relapsed/refractory MCL. Disclosures: Wang: Onyx Pharmaceuticals: Research Funding.

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