RESUMO
A cyclocyanine (CC)-based organic small molecule two-photon (TP) fluorescent probe (CCNa1) was developed for mitochondrial sodium ion sensing. CCNa1 exhibits a low solvatochromic shift and strong TP fluorescence enhancement at 575 nm upon binding to Na+ and is insensitive to other metal ions and to pH. CCNa1 demonstrated fast cell loading ability, biocompatibility, and sensitive response to mitochondrial Na+ influx in live cells and mouse brain tissue.
Assuntos
Corantes Fluorescentes/química , Mitocôndrias/química , Sódio/análise , Animais , Éteres de Coroa/química , Éteres de Coroa/efeitos da radiação , Éteres de Coroa/toxicidade , Corantes Fluorescentes/efeitos da radiação , Corantes Fluorescentes/toxicidade , Células HeLa , Compostos Heterocíclicos de 4 ou mais Anéis/química , Compostos Heterocíclicos de 4 ou mais Anéis/efeitos da radiação , Compostos Heterocíclicos de 4 ou mais Anéis/toxicidade , Hipocampo/metabolismo , Humanos , Camundongos , Fótons , Sódio/metabolismoRESUMO
Diosbulbin-B (DB) is a promising therapeutic drug for cancer treatment; however, DB-induced hepatotoxicity seriously limits its clinical utilization. Based on this, the present study investigated whether the Angelica sinensis extract, angelica sinensis polysaccharide (ASP), was effective to attenuate DB-induced cytotoxicity in hepatocytes. The primary hepatocytes were isolated from rats and cultured in vitro, which were subsequently treated with high-dose DB (100 µM) and ASP (12 µg/ml) to establish the DB-induced hepatotoxicity models. MTT assay and flow cytometry (FCM) were performed to evaluate cell viability, and the results showed that high-dose DB-induced cell apoptosis and inhibition of proliferation were reversed by co-treating cells with ASP, which were supported by our Western Blot assay data that ASP upregulated Cyclin D1 and CDK2 to abrogate high-dose DB-induced cell cycle arrest. In addition, ASP exerted its regulating effects on cell autophagy, and we found that ASP increased LC3B-II/I ratio and Atg5, but decreased p62 to activate the autophagy flux. Of note, the MEK/ERK pathway could be activated by ASP in the DB-treated hepatocytes, and the protective effects of ASP on high-dose DB-induced hepatocyte death were abolished by co-treating cells with the autophagy inhibitor (3-methyladenine, 3-MA) and MEK/ERK selective inhibitor (SCH772984). Moreover, blockage of the MEK/ERK pathway suppressed cell autophagy in the hepatocytes co-treated with ASP and high-dose DB. Taken together, this in vitro study illustrated that ASP activated the MEK/ERK pathway mediated autophagy to suppress high-dose DB-induced hepatotoxicity.
Assuntos
Angelica sinensis/química , Compostos Heterocíclicos de 4 ou mais Anéis/toxicidade , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Extratos Vegetais/farmacologia , Polissacarídeos/farmacologia , Animais , Apoptose/efeitos dos fármacos , Autofagia/efeitos dos fármacos , Sobrevivência Celular , Células Cultivadas , Hepatócitos/efeitos dos fármacos , Hepatócitos/metabolismo , RatosRESUMO
Afidopyropen is an insecticide that acts as a transient receptor potential vanilloid subtype (TRPV) channel modulator in chordotonal organs of target insects and has been assessed for a wide range of toxicity endpoints including chronic toxicity and carcinogenicity in rats and mice. The current study evaluates the toxicokinetic properties of afidopyropen and its plasma metabolites in rats at dose levels where the pharmacokinetics (PK) are linear and nonlinear in an attempt to identify a point of inflection. Based on the results of this study and depending on the analysis method used, the kinetically derived maximum dose (KMD) is estimated to be between 2.5 and 12.5 mg/kg bw/d with linearity observed at doses below 2.5 mg/kg bw/d. A defined point of inflection could not be determined. These data demonstrate that consideration of PK is critical for improving the dose-selection in toxicity studies as well as to enhance human relevance of the interpretation of animal toxicity studies. The study also demonstrates the technical difficulty in obtaining a defined point of inflection from in vivo PK data.
Assuntos
Compostos Heterocíclicos de 4 ou mais Anéis/toxicidade , Inseticidas/toxicidade , Lactonas/toxicidade , Testes de Toxicidade Subaguda/métodos , Administração Oral , Animais , Relação Dose-Resposta a Droga , Feminino , Compostos Heterocíclicos de 4 ou mais Anéis/administração & dosagem , Compostos Heterocíclicos de 4 ou mais Anéis/farmacocinética , Inseticidas/administração & dosagem , Inseticidas/farmacocinética , Lactonas/administração & dosagem , Lactonas/farmacocinética , Masculino , Modelos Animais , Ratos , Organismos Livres de Patógenos Específicos , Testes de Toxicidade , ToxicocinéticaRESUMO
BACKGROUND: The ongoing global malaria eradication campaign requires development of potent, safe, and cost-effective drugs lacking cross-resistance with existing chemotherapies. One critical step in drug development is selecting a suitable clinical candidate from late leads. The process used to select the clinical candidate SJ733 from two potent dihydroisoquinolone (DHIQ) late leads, SJ733 and SJ311, based on their physicochemical, pharmacokinetic (PK), and toxicity profiles is described. METHODS: The compounds were tested to define their physicochemical properties including kinetic and thermodynamic solubility, partition coefficient, permeability, ionization constant, and binding to plasma proteins. Metabolic stability was assessed in both microsomes and hepatocytes derived from mice, rats, dogs, and humans. Cytochrome P450 inhibition was assessed using recombinant human cytochrome enzymes. The pharmacokinetic profiles of single intravenous or oral doses were investigated in mice, rats, and dogs. RESULTS: Although both compounds displayed similar physicochemical properties, SJ733 was more permeable but metabolically less stable than SJ311 in vitro. Single dose PK studies of SJ733 in mice, rats, and dogs demonstrated appreciable oral bioavailability (60-100%), whereas SJ311 had lower oral bioavailability (mice 23%, rats 40%) and higher renal clearance (10-30 fold higher than SJ733 in rats and dogs), suggesting less favorable exposure in humans. SJ311 also displayed a narrower range of dose-proportional exposure, with plasma exposure flattening at doses above 200 mg/kg. CONCLUSION: SJ733 was chosen as the candidate based on a more favorable dose proportionality of exposure and stronger expectation of the ability to justify a strong therapeutic index to regulators.
Assuntos
Antimaláricos/farmacologia , Isoquinolinas/farmacologia , Animais , Antimaláricos/farmacocinética , Antimaláricos/toxicidade , Disponibilidade Biológica , Cães , Hepatócitos/efeitos dos fármacos , Compostos Heterocíclicos de 4 ou mais Anéis/farmacocinética , Compostos Heterocíclicos de 4 ou mais Anéis/farmacologia , Compostos Heterocíclicos de 4 ou mais Anéis/toxicidade , Humanos , Isoquinolinas/farmacocinética , Isoquinolinas/toxicidade , Camundongos , Microssomos Hepáticos/efeitos dos fármacos , RatosRESUMO
Blebbistatin, para-nitroblebbistatin (NBleb), and para-aminoblebbistatin (AmBleb) are highly useful tool compounds as they selectively inhibit the ATPase activity of myosin-2 family proteins. Despite the medical importance of the myosin-2 family as drug targets, chemical optimization has not yet provided a promising lead for drug development because previous structure-activity-relationship studies were limited to a single myosin-2 isoform. Here we evaluated the potential of blebbistatin scaffold for drug development and found that D-ring substitutions can fine-tune isoform specificity, absorption-distribution-metabolism-excretion, and toxicological properties. We defined the inhibitory properties of NBleb and AmBleb on seven different myosin-2 isoforms, which revealed an unexpected potential for isoform specific inhibition. We also found that NBleb metabolizes six times slower than blebbistatin and AmBleb in rats, whereas AmBleb metabolizes two times slower than blebbistatin and NBleb in human, and that AmBleb accumulates in muscle tissues. Moreover, mutagenicity was also greatly reduced in case of AmBleb. These results demonstrate that small substitutions have beneficial functional and pharmacological consequences, which highlight the potential of the blebbistatin scaffold for drug development targeting myosin-2 family proteins and delineate a route for defining the chemical properties of further derivatives to be developed. SIGNIFICANCE STATEMENT: Small substitutions on the blebbistatin scaffold have beneficial functional and pharmacological consequences, highlighting their potential in drug development targeting myosin-2 family proteins.
Assuntos
Absorção Fisico-Química , Descoberta de Drogas , Compostos Heterocíclicos de 4 ou mais Anéis/metabolismo , Compostos Heterocíclicos de 4 ou mais Anéis/farmacologia , Miosinas/antagonistas & inibidores , Animais , Compostos Heterocíclicos de 4 ou mais Anéis/química , Compostos Heterocíclicos de 4 ou mais Anéis/toxicidade , Humanos , Simulação de Dinâmica Molecular , Miosinas/química , Conformação Proteica , Ratos , Distribuição TecidualRESUMO
INTRODUCTION: Colitis-associated cancer (CAC) accounts for approximately 15% of IBD patient mortalities. However, currently available anti-CAC drugs possess many disadvantages including safety, specificity and side effects. Therefore, the development of novel anti-CAC compounds is imperative. HLJ2 was a monomeric compound synthesized by our institute and reported to have an effect on ulcer colitis. METHODS: In vivo the AOM/DSS-induced CAC model was used to evaluate the effects of HLJ2 on ameliorating CAC symptoms, immunohistochemical analysis was used to analyze the pathological damage to colons and epithelial-mesenchymal transition was for changes of cytokines. In vitro, flow cytometric analysis, immunofluorescence and Western blot were used to detect the inhibition effect of HLJ2 on nuclear factor-κB and epithelial-mesenchymal transition in TGF-ß1-stimulated SW480 cells. RESULTS: In the AOM/DSS animal model, HLJ2 was demonstrated to inhibit the secretion of inflammatory cytokines and nuclear factor-κB, levels of tumorigenesis-related proteins including snail, and finally inhibited a key step in metastasis, epithelial-mesenchymal transition. In vitro, HLJ2 was also shown to inhibit nuclear factor-κB and epithelial-mesenchymal transition in TGF-ß1-stimulated SW480 cells in accordance with in vivo results. Meanwhile, the nuclear factor-κB inhibitor could interrupt the effect of HLJ2 on epithelial-mesenchymal transition. DISCUSSION: HLJ2 may ameliorate CAC through inhibiting nuclear factor-κB and then downstream epithelial-mesenchymal transition. The combination of the obvious improvement in effects on CAC without obvious side effects suggests that HLJ2 could be developed as a potential CAC therapeutic candidate.
Assuntos
Antineoplásicos Fitogênicos/uso terapêutico , Colite Ulcerativa/complicações , Neoplasias do Colo/tratamento farmacológico , Transição Epitelial-Mesenquimal/efeitos dos fármacos , Compostos Heterocíclicos de 4 ou mais Anéis/uso terapêutico , NF-kappa B/antagonistas & inibidores , Animais , Antineoplásicos Fitogênicos/toxicidade , Azoximetano , Linhagem Celular Tumoral , Colite Ulcerativa/induzido quimicamente , Colite Ulcerativa/patologia , Colo/patologia , Neoplasias do Colo/etiologia , Neoplasias do Colo/patologia , Citocinas/metabolismo , Sulfato de Dextrana , Feminino , Compostos Heterocíclicos de 4 ou mais Anéis/toxicidade , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Fator de Crescimento Transformador beta1/farmacologiaRESUMO
The rhizome of Dioscorea bulbifera L. (DBL) is a popular traditional herb in the treatment of goiters, breast lumps, and tumors. Unfortunately, DBL can give rise to severe hepatotoxicity. More than 100 cases of liver injury, due to the usage of DBL in China, have been reported in the past half-century. The main toxic components of DBL are furanoditerpenoids diosbulbin B (DSB) as well as 8-epidiosbulbin E (EEA). This toxic effect requires metabolic oxidation of the furan ring mediated by cytochrome P450 enzymes, and the P450 3A subfamily is the main enzyme responsible for the reported hepatotoxicity. cis-Enedial intermediates resulting from furan ring oxidation can react with nucleophilic sites of macromolecules, such as protein and DNA, which may trigger the toxicities. This review illustrates the liver injury induced by DBL including metabolic activation of DSB and EEA, the essential components responsible for DBL-induced hepatotoxicity, along with biochemical mechanisms of their toxic actions. It will facilitate the development of approaches to prevent and intervene in liver injury induced by DBL for its safe use in clinical practice.
Assuntos
Ativação Metabólica , Doença Hepática Induzida por Substâncias e Drogas , Dioscorea/química , Diterpenos/toxicidade , Compostos Heterocíclicos de 4 ou mais Anéis/toxicidade , Animais , Sistema Enzimático do Citocromo P-450/metabolismo , Diterpenos/metabolismo , Medicamentos de Ervas Chinesas , Compostos Heterocíclicos de 4 ou mais Anéis/metabolismo , Humanos , Fígado/efeitos dos fármacos , Fígado/enzimologia , Fígado/metabolismoRESUMO
The tight junctions between capillary endothelial cells of the blood-brain barrier (BBB) restricts the entry of therapeutics into the brain. Potential of the intranasal delivery tool has been explored in administering the therapeutics directly to the brain, thus bypassing BBB. The objective of this study was to develop and optimize an intranasal mucoadhesive nanoemulsion (MNE) of asenapine maleate (ASP) in order to enhance the nasomucosal adhesion and direct brain targetability for improved efficacy and safety. Box-Behnken statistical design was used to recognize the crucial formulation variables influencing droplet size, size distribution and surface charge of ASP-NE. ASP-MNE was obtained by incorporating GRAS mucoadhesive polymer, Carbopol 971 in the optimized NE. Optimized ASP-MNE displayed spherical morphology with a droplet size of 21.2 ± 0.15 nm and 0.355 polydispersity index. Improved ex-vivo permeation was observed in ASP-NE and ASP-MNE, compared to the ASP-solution. Finally, the optimized formulation was found to be safe in ex-vivo ciliotoxicity study on sheep nasal mucosa. The single-dose pharmacokinetic study in male Wistar rats revealed a significant increase in concentration of ASP in the brain upon intranasal administration of ASP-MNE, with a maximum of 284.33 ± 5.5 ng/mL. The time required to reach maximum brain concentration (1 h) was reduced compared to intravenous administration of ASP-NE (3 h). Furthermore, it has been established during the course of present study, that the brain targeting capability of ASP via intranasal administration had enhanced drug-targeting efficiency and drug-targeting potential. In the animal behavioral studies, no extrapyramidal symptoms were observed after intranasal administration of ASP-MNE, while good locomotor activity and hind-limb retraction test established its antipsychotic activity in treated animals. Thus, it can be concluded that the developed intranasal ASP-MNE could be used as an effective and safe tool for brain targeting of ASP in the treatment of psychotic disorders.
Assuntos
Encéfalo/metabolismo , Sistemas de Liberação de Medicamentos , Compostos Heterocíclicos de 4 ou mais Anéis/administração & dosagem , Nanopartículas , Adesividade , Administração Intranasal , Animais , Antipsicóticos/administração & dosagem , Antipsicóticos/farmacocinética , Antipsicóticos/toxicidade , Barreira Hematoencefálica/metabolismo , Dibenzocicloeptenos , Emulsões , Células Endoteliais/metabolismo , Compostos Heterocíclicos de 4 ou mais Anéis/farmacocinética , Compostos Heterocíclicos de 4 ou mais Anéis/toxicidade , Masculino , Mucosa Nasal/metabolismo , Tamanho da Partícula , Polímeros/química , Ratos , Ratos Wistar , Ovinos , Distribuição TecidualRESUMO
Cardiac constructs fabricated using human induced pluripotent stem cells-derived cardiomyocytes (iPSCs-CMs) are useful for evaluating the cardiotoxicity of and cardiac response to new drugs. Previously, we fabricated scaffold-free three-dimensional (3D) tubular cardiac constructs using a bio-3D printer, which can load cardiac spheroids onto a needle array. In this study, we developed a method to measure the contractile force and to evaluate the drug response in cardiac constructs. Specifically, we measured the movement of the needle tip upon contraction of the cardiac constructs on the needle array. The contractile force and beating rate of the cardiac constructs were evaluated by analysing changes in the movement of the needle tip. To evaluate the drug response, contractile properties were measured following treatment with isoproterenol, propranolol, or blebbistatin, in which the movement of the needle tip was increased following isoproterenol treatment, but was decreased following propranolol or blebbistain, treatments. To evaluate cardiotoxicity, contraction and cell viability of the cardiac constructs were measured following doxorubicin treatment. Cell viability was found to decrease with decreasing movement of the needle tip following doxorubicin treatment. Collectively, our results show that this method can aid in evaluating the contractile force of cardiac constructs.
Assuntos
Cardiotoxicidade , Avaliação Pré-Clínica de Medicamentos/métodos , Células-Tronco Pluripotentes Induzidas , Contração Miocárdica/efeitos dos fármacos , Miócitos Cardíacos/efeitos dos fármacos , Impressão Tridimensional , Engenharia Tecidual/métodos , Testes de Toxicidade/métodos , Sobrevivência Celular/efeitos dos fármacos , Doxorrubicina/farmacologia , Doxorrubicina/toxicidade , Compostos Heterocíclicos de 4 ou mais Anéis/farmacologia , Compostos Heterocíclicos de 4 ou mais Anéis/toxicidade , Humanos , Isoproterenol/farmacologia , Isoproterenol/toxicidade , Propranolol/farmacologia , Propranolol/toxicidade , Alicerces TeciduaisRESUMO
Diosbulbin B (DIOB) is an effective component of air potato yam with antitumor and anti-inflammatory activities, and it is the main toxic component leading to hepatotoxicity. However, the mechanism of its hepatotoxicity remains unclear. In this study, we aimed to systematically elucidate the molecular action of DIOB on liver metabolic function through systems toxicology approaches. C57BL/6 mice were orally treated with DIOB (10, 30, 60 mg/kg) for 28 days, and the liver metabonomics and histopathology, molecular docking, mRNA expression levels, and activities of enzymes were analyzed. The results illustrated that DIOB could affect fatty acid and glucose metabolism, block the TCA cycle, and DIOB also could disorder bile acid synthesis and transport and promote the occurrence of hyperbilirubinemia. In addition, DIOB increased Cyp3a11 expression in a dose-dependent manner. Thus, these results provide new insights into the mechanism of hepatotoxicity caused by DIOB.
Assuntos
Doença Hepática Induzida por Substâncias e Drogas/metabolismo , Compostos Heterocíclicos de 4 ou mais Anéis/toxicidade , Fígado/efeitos dos fármacos , Adenosina Trifosfatases/metabolismo , Animais , Doença Hepática Induzida por Substâncias e Drogas/genética , Doença Hepática Induzida por Substâncias e Drogas/patologia , Citocromo P-450 CYP3A/genética , Ácidos Graxos/metabolismo , Glucose/metabolismo , Fígado/metabolismo , Fígado/patologia , Masculino , Proteínas de Membrana/genética , Metabolômica , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos ICR , Mitocôndrias Hepáticas/efeitos dos fármacos , Mitocôndrias Hepáticas/metabolismo , Simulação de Acoplamento Molecular , Biologia de Sistemas , ToxicologiaRESUMO
Diosbulbin-B (DB) was the main compound of Dioscorea bulbifera L, which was widely used for cancer treatment in Asia. However, the hepatotoxicity induced by high-dose DB seriously limited its possibility using for gastric cancer (GC) treatment in clinic. In this study, we found that DB inhibited GC cells and hepatocytes cell viability in a dose- and time-dependent manner. Specifically, high-dose DB (50µM) significantly inhibited cell proliferation and promoted cell apoptosis, while low dose DB (12.5µM) had little effects on cell viability. Besides, high-dose DB (50µM) significantly decreased CircRNA CDR1as levels in gastric cancer cells instead of hepatocytes. Notably, knock-down of CircRNA CDR1as triggered low-dose DB (12.5µM) induced GC cell death, but had little effects on hepatocytes proliferation and apoptosis. Further results showed that CircRNA CDR1as increased REGγ expressions in GC cells by sponging miR-7-5p, and high-dose DB (50µM) increased miR-7-5p levels and inhibited REGγ expressions in GC cells instead of hepatocytes. In addition, either downregulated miR-7-5p or overexpressed REGγ reversed the promoting effects of downregulated CircRNA CDR1as on low-dose DB-induced GC cell death. Taken together, we concluded that knock-down of CircRNA CDR1as specifically promoted the cytotoxic effects of low-dose DB on GC cells instead of hepatocytes by regulating miR-7-5p/REGγ axis.
Assuntos
Compostos Heterocíclicos de 4 ou mais Anéis/toxicidade , Compostos Heterocíclicos de 4 ou mais Anéis/uso terapêutico , MicroRNAs/metabolismo , RNA Circular/metabolismo , Neoplasias Gástricas/tratamento farmacológico , Autoantígenos/metabolismo , Morte Celular , Linhagem Celular Tumoral , Proliferação de Células , Regulação para Baixo , Células HEK293 , Hepatócitos , Humanos , Complexo de Endopeptidases do Proteassoma/metabolismo , RNA Circular/genética , Neoplasias Gástricas/metabolismoRESUMO
Bictegravir (BIC), a newly FDA-approved integrase strand transfer inhibitor (INSTI), as a single tablet regimen has proven efficacious in treating HIV-1 and SIV viruses, with reduced resistance. BIC clinical trials have not investigated its prophylaxis potency. This study investigates the HIV prevention potency of a novel long-acting BIC nano-formulation aimed to improve adherence. Poly (lactic-co-glycolic acid) loaded BIC nanoparticles (BIC NPs) were formulated using an oil-in-water emulsion methodology. BIC NPs were <200â¯nm in size, with 47.9⯱â¯6.9% encapsulation efficiency. A novel, sensitive and high throughput LC-MS/MS method was used to estimate intracellular pharmacokinetics (PK) of BIC NPs and compared to BIC solution demonstrated prolonged intracellular BIC retention. BIC NPs safety was assessed based on cytotoxicity. Further, in-vitro prevention study of BIC NPs vs BIC solution was assessed against HIV-1NLX and HIV-1ADA on TZM-bl cell line and PBMCs, respectively. BIC nanoencapsulation demonstrated elevated cellular cytotoxicity concentration (CC50: 2.25⯵M (BIC solution) to 820.4⯵M (BIC NPs)] and lowers HIV-1 inhibitory concentration [EC50: 0.604⯵M (BIC solution) to 0.0038⯵M (BIC NPs)) thereby improving selectivity index (SI) from 3.7 (BIC solution) to 215,789 (BIC NP) for TZM-bl cells. Comparable results in PBMCs were obtained where BIC NPs improved SI from 0.29 (BIC solution) to 523.33 (BIC NPs). This demonstrates long-acting BIC nano-formulation with sustained drug-release potency, improved BIC cytotoxicity and enhanced HIV-1 protection compared to BIC in solution.
Assuntos
Sistemas de Liberação de Medicamentos/métodos , Infecções por HIV/tratamento farmacológico , Compostos Heterocíclicos de 4 ou mais Anéis/farmacocinética , Nanopartículas/virologia , Amidas , Linhagem Celular , HIV-1/efeitos dos fármacos , Compostos Heterocíclicos com 3 Anéis , Compostos Heterocíclicos de 4 ou mais Anéis/toxicidade , Humanos , Concentração Inibidora 50 , Nanopartículas/uso terapêutico , Piperazinas , Estudo de Prova de Conceito , PiridonasRESUMO
BACKGROUND: Toosendan Fructus is traditionally used as an insecticide or digestive tract parasiticide for treating digestive parasites in China. It is recorded to have little toxicity in Chinese Pharmacopoeia and has been found to cause severe liver injury during clinical practice. PURPOSE: This study aims to identify candidate serum microRNAs (miRNAs) as potential toxicological biomarkers for reflecting the hepatotoxicity induced by toosendanin (TSN), which is the main toxic compound isolated from Toosendan Fructus METHODS: Alanine/aspartate aminotransferase (ALT/AST) activities detection and liver histological observation were performed to evaluate the liver injury induced by TSN or other hepatotoxicants in mice. miRNAs chip analysis and Real-time PCR assay were conducted to identify the altered miRNAs in serum from TSN-treated mice RESULTS: The results of serum ALT/AST and liver histological evaluation showed that TSN (10â¯mg/kg) induced hepatotoxicity in mice. The results of miRNAs chip showed that the expression of 81 serum miRNAs was obviously altered in mice treated with TSN for 12â¯h, and 22 of them have passed the further validation in serum from mice treated with TSN for both 6â¯h and 12â¯h. These 22 miRNAs were supposed to be the candidate toxicological biomarkers for TSN-induced hepatotoxicity with more sensitivity as compared to the alteration of AST or ALT activity. Moreover, the expression of miRNA-122-3p and mcmv-miRNA-m01-4-3p was not only increased in TSN-treated mice, but also increased in mice treated with other hepatotoxicants including acetaminophen (APAP), monocrotaline (MCT) and diosbuibin B (DB). Only the expression of serum miRNA-367-3p was increased in TSN-treated mice but not changed in the liver injury induced by APAP, MCT or DB CONCLUSION: miR-122-3p and mcmv-miRNA-m01-4-3p may be two commonly sensitive biomarkers for reflecting the hepatotoxicity induced by exogenous hepatotoxicants, and miR-367-3p may be a specific biomarker for reflecting the liver injury induced by TSN.
Assuntos
Doença Hepática Induzida por Substâncias e Drogas/sangue , Medicamentos de Ervas Chinesas/toxicidade , Medicina Tradicional Chinesa/efeitos adversos , MicroRNAs/sangue , Acetaminofen/toxicidade , Alanina Transaminase/metabolismo , Animais , Aspartato Aminotransferases/metabolismo , Biomarcadores/sangue , Compostos Heterocíclicos de 4 ou mais Anéis/toxicidade , Fígado/efeitos dos fármacos , Fígado/enzimologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Monocrotalina/toxicidade , Distribuição Aleatória , Organismos Livres de Patógenos EspecíficosRESUMO
Chemotherapeutic agents generally suffer from off-target cytotoxicity in noncancerous cell types, leading to undesired side effects. As a result, significant effort has been put into identifying compounds that are selective for cancerous over noncancerous cell types. Our laboratory has recently developed a series of near-infrared (NIR) fluorophores containing a phosphinate functionality at the bridging position of a xanthene scaffold, termed Nebraska Red (NR) fluorophores. Herein, we report the selective cytotoxicity of one NR derivative, NR744 , against HeLa (cervical cancer) cells versus NIH-3T3 (noncancerous fibroblast) cells. Mechanistic studies based on the NIR fluorescence signal of NR744 showed distinct subcellular localization in HeLa (mitochondrial) versus NIH-3T3 (lysosomal) that resulted from the elevated mitochondrial potential in HeLa cells. This study provides a new, NIR scaffold for the further development of reagents for targeted cancer therapy.
Assuntos
Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Corantes Fluorescentes/farmacologia , Ácidos Fosfínicos/farmacologia , Animais , Antineoplásicos/metabolismo , Antineoplásicos/toxicidade , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Corantes Fluorescentes/metabolismo , Corantes Fluorescentes/toxicidade , Compostos Heterocíclicos com 3 Anéis/metabolismo , Compostos Heterocíclicos com 3 Anéis/farmacologia , Compostos Heterocíclicos com 3 Anéis/toxicidade , Compostos Heterocíclicos de 4 ou mais Anéis/metabolismo , Compostos Heterocíclicos de 4 ou mais Anéis/farmacologia , Compostos Heterocíclicos de 4 ou mais Anéis/toxicidade , Humanos , Camundongos , Microscopia Confocal/métodos , Microscopia de Fluorescência/métodos , Mitocôndrias/metabolismo , Células NIH 3T3 , Ácidos Fosfínicos/metabolismo , Ácidos Fosfínicos/toxicidadeRESUMO
Diosbulbin B (DB) is the main hepatotoxic compound in Airpotato yam, which is traditionally used for treating thyroid disease and cancer in China, and its hepatotoxic mechanism still remains unclear. This study aims to investigate its hepatotoxic mechanism by focusing on regulating microRNA (miRNA). DB induced hepatotoxicity both in vivo and in vitro. Results of miRNA chip analysis showed that the expression of eleven miRNAs was up-regulated and twelve miRNAs was down-regulated in livers from DB-treated mice. The altered expression of seven miRNAs was further validated by using real-time polymerase chain reaction (RT-PCR) assay. DB induced G2/M arrest in L-02/cytochrome P450 3A4 (CYP3A4) cells in both concentration- and time-dependent manner. A total of eleven predicted target genes was related with cell cycle regulation of those above seven miRNAs, among which the mRNA and protein expression of cyclin-dependent kinase 1 (CDK1) decreased both in vivo and in vitro. Both miR-378a-5p and miR-186-3p have binding sites in the 3'-untranslated region (UTR) of CDK1. With the use of CDK1 3'-UTR luciferase reporter assay, miR-378a-5p and miR-186-3p was found to down-regulate the luciferase activity. The mimics of miR-378a-5p or miR-186-3p reduced CDK1 expression in L-02/CYP3A4 cells, but their inhibitors reversed the decreased CDK1 expression induced by DB. Moreover, overexpression of miR-186-3p inhibitor reversed the G2/M cell cycle arrest induced by DB in L-02/CYP3A4 cells. Taken together, our results showed that DB induced hepatotoxicity by inducing G2/M cell cycle arrest in hepatocytes via miR-186-3p or miR-378a-5p-mediated the reduced CDK1 expression.
Assuntos
Pontos de Checagem da Fase G2 do Ciclo Celular/efeitos dos fármacos , Hepatócitos/efeitos dos fármacos , Compostos Heterocíclicos de 4 ou mais Anéis/toxicidade , MicroRNAs/metabolismo , Animais , Proteína Quinase CDC2 , Linhagem Celular , Citocromo P-450 CYP3A/genética , Citocromo P-450 CYP3A/metabolismo , Relação Dose-Resposta a Droga , Regulação da Expressão Gênica/efeitos dos fármacos , Hepatócitos/citologia , Hepatócitos/metabolismo , Compostos Heterocíclicos de 4 ou mais Anéis/administração & dosagem , Humanos , Camundongos , Camundongos Endogâmicos ICR , MicroRNAs/genética , Distribuição Aleatória , Transcriptoma/efeitos dos fármacosRESUMO
Afidopyropen is a novel insecticide that acts as a TRPV channel modulator in chordotonal organs of target insects. In two carcinogenicity studies with Fischer rats, an increased incidence of uterine adenocarcinomas was observed at 1000 and 3000â¯ppm. This finding prompted an investigation of the mechanism of the tumor formation as well as the relevance of this mechanism to humans. The mechanistic work took parallel paths: one path investigated the pharmacokinetic properties of the test substance at the doses where the tumors were found; while the second path examined the key mechanistic events that culminated in uterine adenocarcinomas. The results of the investigation indicated that the tumors only occurred at doses where excretion of test substance was saturated - indicating that homeostatic biological and/or physiological processes were overwhelmed. At the doses where these processes were overwhelmed, the test substance acted through a mechanism of dopamine agonism, triggering a cascade key events that resulted in uterine adenocarcinomas. An analysis of these mechanisms observed in rat showed that they are both quantitatively (pharmacokinetic mechanism) and qualitatively (dopamine agonism mechanism) not relevant to humans. Therefore the uterine adenocarcinomas observed in the rat associated with high doses of Afidopyropen are not expected to pose a carcinogenic risk to humans.
Assuntos
Adenocarcinoma/induzido quimicamente , Carcinógenos/toxicidade , Agonistas de Dopamina/toxicidade , Compostos Heterocíclicos de 4 ou mais Anéis/toxicidade , Inseticidas/toxicidade , Lactonas/toxicidade , Neoplasias Uterinas/induzido quimicamente , Animais , Carcinógenos/farmacocinética , Progressão da Doença , Agonistas de Dopamina/farmacocinética , Feminino , Compostos Heterocíclicos de 4 ou mais Anéis/farmacocinética , Humanos , Inseticidas/farmacocinética , Lactonas/farmacocinética , Masculino , Ratos Endogâmicos F344 , Medição de Risco , Testes de ToxicidadeRESUMO
Penicillium roqueforti is a common food and feed contaminant. However, it is also worldwide renowned for its use as a technological culture responsible for the typicity of blue-veined cheese. Members of the P. roqueforti species are also known to be able to produce secondary metabolites including mycophenolic acid (MPA) and roquefortine C (ROQ C) mycotoxins. In order to more closely simulate the reality of mycotoxin exposure through contaminated food consumption, this work investigated the toxicological effects of MPA and ROQ C not only in acute but also in chronic (i.e. 21-days continuous exposure) conditions on Caco-2 cells. Acute exposure to high MPA or ROQ C concentrations induced an increase of IL-8 secretion. Effects of 21-days continuous exposure on barrier integrity, based on concentrations found in blue-veined cheese and mean of blue cheese intake by French consumers, were monitored. Concerning exposure to ROQ C, no alteration of the intestinal barrier was observed. In contrast, the highest tested MPA concentration (780⯵M) induced a decrease in the barrier function of Caco-2 cell monolayers, but no paracellular passage of bacteria was observed. This study highlighted that exposure to MPA and ROQ C average concentrations found in blue-veined cheese does not seem to induce significant toxicological effects in the tested conditions.
Assuntos
Micotoxinas/toxicidade , Penicillium/química , Doença Aguda , Fosfatase Alcalina/metabolismo , Translocação Bacteriana/efeitos dos fármacos , Células CACO-2 , Queijo/microbiologia , Doença Crônica , Enterócitos/efeitos dos fármacos , Compostos Heterocíclicos de 4 ou mais Anéis/toxicidade , Humanos , Indóis/toxicidade , Interleucina-8/metabolismo , Mitocôndrias/efeitos dos fármacos , Ácido Micofenólico/análise , Ácido Micofenólico/toxicidade , Piperazinas/toxicidadeRESUMO
Graphene oxide (GO) is a promising material for biomedical applications, particularly in drug delivery, due to its exceptional chemical and physical properties. In this work, an innovative GO-based carrier was developed by modifying GO with chitosan (CHI) to improve the biocompatibility, and followed by the conjugation of hyaluronic acid (HA), the target ligand for CD44, to realize the specific recognition of tumor cells and improve the efficiency of anti-tumor drug delivery. The resulting product GO-CHI-HA was loaded with an anti-cancer drug SNX-2112, which is the Hsp90 inhibitor. The total release amount and release rate of SNX-2112 were significantly higher in acidic condition than in physiological condition. GO-CHI-HA with a low concentration had little impact on the lysis of red blood cells (RBCs) and blood coagulation and showed low toxicity in A549 cells and NHBE cells. The GO-CHI-HA/SNX-2112 proved to be effective in inhibiting and killing A549 cells while having lower cytotoxicity against normal human bronchial epithelial cells (NHBE cells). Furthermore, in vivo toxicity of the materials towards vital organs in SD rats were also studied through histological examinations and blood property analyses, the results of which showed that although inflammatory response was developed in the short-term, GO-CHI-HA/SNX-2112 caused no severe long-term injury. Therefore, this drug delivery system showed great potential as an effective and safe drug delivery system with little adverse side effects for cancer therapy.
Assuntos
Antineoplásicos/administração & dosagem , Portadores de Fármacos/química , Grafite/química , Compostos Heterocíclicos de 4 ou mais Anéis/administração & dosagem , Neoplasias Pulmonares/tratamento farmacológico , Nanocompostos/química , Polissacarídeos/química , Animais , Antineoplásicos/uso terapêutico , Antineoplásicos/toxicidade , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Portadores de Fármacos/efeitos adversos , Hemólise/efeitos dos fármacos , Compostos Heterocíclicos de 4 ou mais Anéis/uso terapêutico , Compostos Heterocíclicos de 4 ou mais Anéis/toxicidade , Humanos , Masculino , Nanocompostos/efeitos adversos , Ratos , Ratos Sprague-DawleyRESUMO
Due to extremely chemical complexity, identification of potential toxicity-related constituents from an herbal medicine (HM) still remains challenging. Traditional toxicity-guided separation procedure suffers from time- and labor-consumption and neglects the additive effect of multi-components. In this study, we proposed a screening strategy called "hepatotoxic equivalent combinatorial markers (HECMs)" for a hepatotoxic HM, Dioscorea bulbifera tuber (DBT). Firstly, the chemical constituents in DBT extract were globally characterized. Secondly, the fingerprints of DBT extracts were established and their in vivo hepatotoxicities were tested. Thirdly, three chemometric tools including partial least squares regression (PLSR), back propagation-artificial neural network (BP-ANN) and cluster analysis were applied to model the fingerprint-hepatotoxicity relationship and to screen hepatotoxicity-related markers. Finally, the chemical combination of markers was subjected to hepatotoxic equivalence evaluation. A total of 40 compounds were detected or tentatively characterized. Two diterpenoid lactones, 8-epidiosbulbin E acetate (EEA) and diosbulbin B (DIOB), were discovered as the most hepatotoxicity-related markers. The chemical combination of EEA and DIOB, reflecting the whole hepatotoxicity of original DBT extract with considerable confidential interval, was verified as HECMs for DBT. The present study is expected not only to efficiently discover hepatotoxicity-related markers of HMs, but also to rationally evaluate/predict the hepatotoxicity of HMs.
Assuntos
Dioscorea/química , Medicamentos de Ervas Chinesas/análise , Medicamentos de Ervas Chinesas/toxicidade , Fígado/efeitos dos fármacos , Animais , Biomarcadores/metabolismo , Análise por Conglomerados , Diterpenos/toxicidade , Medicamentos de Ervas Chinesas/administração & dosagem , Compostos Heterocíclicos de 4 ou mais Anéis/toxicidade , Análise dos Mínimos Quadrados , Fígado/metabolismo , Camundongos , Redes Neurais de Computação , Tubérculos/química , Testes de ToxicidadeRESUMO
Lack of novel antifungal agents and severe drug resistance have led to high incidence and associated mortality of invasive fungal infections. To tackle the challenges, novel antifungal agents with new chemotype, fungicidal activity and anti-resistant potency are highly desirable. On the basis of our previously identified simplified analogue of antifungal natural product sampangine, systemic structure-activity relationships were clarified and two novel derivatives showed promising features as novel antifungal lead compounds. Compounds 22b and 22c showed good fungicidal activity against both fluconazole-sensitive and fluconazole-resistant Candida albicans strains. Moreover, they were proven to be potent inhibitors of Candida albicans biofilm formation and yeast-to-hypha morphological transition by down-regulating biofilm-associated genes. In a rat vaginal Candida albicans infection model, compounds 22b and 22c showed excellent therapeutic effects with low toxicity. The results highlighted the potential of sampangine derivatives to overcome fluconazole-related and biofilm-related drug resistance.