ABSTRACT
Gingiva hyperpigmentation resulting from physiological melanosis causes aesthetic discomfort and is usually perceived as a disease by patients because healthy attached gingiva is typically characterized by coral pink coloring with stippling and scalloped contours. When physiological melanosis compromises the aesthetics of smiling, it may induce insecurity in patients, who usually seek out alternatives for reducing or eliminating hyperpigmentation. We present a case report of a surgical procedure combining gingivectomy with gingivoplasty for the management of physiological melanosis. The surgical procedure was performed on a 40-year-old female patient with bilateral pigmentation in both arches. The results of the histological analysis confirm the diagnoses of melanotic macula, with papillary hyperplasia and cytopathic changes being suggestive of HPV infection, which was verified using an immunohistochemistry analysis based on the detection of a major capsid protein of HPV. Acceptable functional and aesthetic results were obtained for the patient without major discomfort during the postoperative period. In cases when HPV infection is present, long-term follow-up becomes necessary.
ABSTRACT
Ilama leaves are an important source of secondary metabolites with promising anticancer properties. Cancer is a disease that affects a great number of people worldwide. This work aimed to investigate the in vivo, in vitro and in silico anticancer properties of three acyclic terpenoids (geranylgeraniol, phytol and farnesyl acetate) isolated from petroleum ether extract of ilama leaves. Their cytotoxic activity against U-937 cells was assessed using flow cytometry to determine the type of cell death and production of reactive oxygen species (ROS). Also, a morphological analysis of the lymph nodes and a molecular docking study using three proteins related with cancer as targets, namely, Bcl-2, Mcl-1 and VEGFR-2, were performed. The flow cytometry and histomorphological analysis revealed that geranylgeraniol, phytol and farnesyl acetate induced the death of U-937 cells by late apoptosis and necrosis. Geranylgeraniol and phytol induced a significant increase in ROS production. The molecular docking studies showed that geranylgeraniol had more affinity for Bcl-2 and VEGFR-2. In the case of farnesyl acetate, it showed the best affinity for Mcl-1. This study provides information that supports the anticancer potential of geranylgeraniol, phytol and farnesyl acetate as compounds for the treatment of cancer, particularly with the potential to treat non-Hodgkin's lymphoma.
Subject(s)
Molecular Docking Simulation , Plant Extracts , Plant Leaves , Plants, Medicinal , Reactive Oxygen Species , Humans , Plant Leaves/chemistry , Plants, Medicinal/chemistry , Plant Extracts/chemistry , Plant Extracts/pharmacology , Reactive Oxygen Species/metabolism , Antineoplastic Agents, Phytogenic/pharmacology , Antineoplastic Agents, Phytogenic/chemistry , Mexico , Apoptosis/drug effects , Cell Line, Tumor , Animals , Vascular Endothelial Growth Factor Receptor-2/metabolism , Vascular Endothelial Growth Factor Receptor-2/antagonists & inhibitors , Computer Simulation , Proto-Oncogene Proteins c-bcl-2/metabolism , U937 CellsABSTRACT
Odontogenic keratocyst (OK) is a benign intraosseous cystic lesion characterized by a parakeratinized stratified squamous epithelial lining with palisade basal cells. It represents 10-12% of odontogenic cysts. The changes in its classification as a tumor or cyst have increased interest in its pathogenesis. OBJECTIVE: Identify key genes in the pathogenesis of sporadic OK through in silico analysis. MATERIALS AND METHODS: The GSE38494 technical sheet on OK was analyzed using GEOR2. Their functional and canonical signaling pathways were enriched in the NIH-DAVID bioinformatic platform. The protein-protein interaction network was constructed by STRING and analyzed with Cytoscape-MCODE software v 3.8.2 (score > 4). Post-enrichment analysis was performed by Cytoscape-ClueGO. RESULTS: A total of 768 differentially expressed genes (DEG) with a fold change (FC) greater than 2 and 469 DEG with an FC less than 2 were identified. In the post-enrichment analysis of upregulated genes, significance was observed in criteria related to the organization of the extracellular matrix, collagen fibers, and endodermal differentiation, while the downregulated genes were related to defensive response mechanisms against viruses and interferon-gamma activation. CONCLUSIONS: Our in silico analysis showed a significant relationship with mechanisms of extracellular matrix organization, interferon-gamma activation, and response to viral infections, which must be validated through molecular assays.
Subject(s)
Odontogenic Cysts , Odontogenic Tumors , Humans , Interferon-gamma , Odontogenic Cysts/genetics , Odontogenic Cysts/pathology , Odontogenic Tumors/pathology , Protein Interaction Maps/geneticsABSTRACT
Cell proliferation and invasion are characteristic of many tumors, including ameloblastoma, and are important features to target in possible future therapeutic applications. OBJECTIVE: The objective of this study was the identification of key genes and inhibitory drugs related to the cell proliferation and invasion of ameloblastoma using bioinformatic analysis. METHODS: The H10KA_07_38 gene profile database was analyzed by Rstudio and ShinyGO Gene Ontology enrichment. String, Cytoscape-MCODE, and Kaplan-Meier plots were generated, which were subsequently validated by RT-qPCR relative expression and immunoexpression analyses. To propose specific inhibitory drugs, a bioinformatic search using Drug Gene Budger and DrugBank was performed. RESULTS: A total of 204 significantly upregulated genes were identified. Gene ontology enrichment analysis identified four pathways related to cell proliferation and cell invasion. A total of 37 genes were involved in these pathways, and 11 genes showed an MCODE score of ≥0.4; however, only SLC6A3, SOX10, and LRP5 were negatively associated with overall survival (HR = 1.49 (p = 0.0072), HR = 1.55 (p = 0.0018), and HR = 1.38 (p = 0.025), respectively). The RT-qPCR results confirmed the significant differences in expression, with overexpression of >2 for SLC6A3 and SOX10. The immunoexpression analysis indicated positive LRP5 and SLC6A3 expression. The inhibitory drugs bioinformatically obtained for the above three genes were parthenolide and vorinostat. CONCLUSIONS: We identify LRP5, SLC6A3, and SOX10 as potentially important genes related to cell proliferation and invasion in the pathogenesis of ameloblastomas, along with both parthenolide and vorinostat as inhibitory drugs that could be further investigated for the development of novel therapeutic approaches against ameloblastoma.
Subject(s)
Ameloblastoma , Humans , Ameloblastoma/genetics , Vorinostat , Cell Proliferation/genetics , Computational Biology , SOXE Transcription Factors/genetics , Low Density Lipoprotein Receptor-Related Protein-5 , Dopamine Plasma Membrane Transport ProteinsABSTRACT
BACKGROUND: Oral cancer has a high prevalence worldwide, and this disease is caused by genetic, immunological, and environmental factors. The main risk factors associated with oral cancer are smoking and alcohol. RESULTS: There are various strategies to reduce risk factors, including prevention programs as well as the consumption of an adequate diet that includes phytochemical compounds derived from cranberries (Vaccinium macrocarpon A.) and blueberries (Vaccinium corymbosum L.); these compounds exhibit antitumor properties. RESULTS: The main outcome of this review is as follows: the properties of phytochemicals derived from cranberries were evaluated for protection against risk factors associated with oral cancer. CONCLUSIONS: The secondary metabolites of cranberries promote biological effects that provide protection against smoking and alcoholism. An alternative for the prevention of oral cancer can be the consumption of these cranberries and blueberries.
ABSTRACT
The antihyperglycemic activity of ethanolic extract from Annona cherimola Miller (EEAch) and its products were evaluated using in vivo and in silico assays. An α-glucosidase inhibition was evaluated with oral sucrose tolerance tests (OSTT) and molecular docking studies using acarbose as the control. SGLT1 inhibition was evaluated with an oral glucose tolerance test (OGTT) and molecular docking studies using canagliflozin as the control. Among all products tested, EEAc, the aqueous residual fraction (AcRFr), rutin, and myricetin reduced the hyperglycemia in DM2 mice. During the carbohydrate tolerance tests, all the treatments reduced the postprandial peak such as the control drugs. In the molecular docking studies, rutin showed more affinity in inhibiting α-glucosidase enzymes and myricetin in inhibiting the SGLT1 cotransporter, showing ∆G values of -6.03 and -3.32 kcal/mol-1, respectively, in α-glucosidase enzymes. In the case of the SGLT1 cotransporter, molecular docking showed ∆G values of 22.82 and -7.89 in rutin and myricetin, respectively. This research sorts in vivo and in silico pharmacological studies regarding the use of A. cherimola leaves as a source for the development of new potential antidiabetic agents for T2D control, such as flavonoids rutin and myricetin.
ABSTRACT
Linearolactone (LL) is a neo-clerodane type diterpene that has been shown to exert giardicidal effects; however, its mechanism of action is unknown. This work analyzes the cytotoxic effect of LL on Giardia intestinalis trophozoites and identifies proteins that could be targeted by this active natural product. Increasing concentrations of LL and albendazole (ABZ) were used as test and reference drugs, respectively. Cell cycle progression, determination of reactive oxygen species (ROS) and apoptosis/necrosis events were evaluated by flow cytometry (FCM). Ultrastructural alterations were analyzed by transmission electron microscopy (TEM). Ligand-protein docking analyses were carried out using the LL structure raised from a drug library and the crystal structure of an aldose reductase homologue (GdAldRed) from G. intestinalis. LL induced partial arrest at the S phase of trophozoite cell cycle without evidence of ROS production. LL induced pronecrotic death in addition to inducing ultrastructural alterations as changes in vacuole abundances, appearance of perinuclear and periplasmic spaces, and deposition of glycogen granules. On the other hand, the in silico study predicted that GdAldRed is a likely target of LL because it showed a favored change in Gibbs free energy for this complex.
ABSTRACT
Heliangolide-type sesquiterpene lactones (HTSLs) are phytocompounds with several pharmacological activities including cytotoxic and antitumor activity. Both bioactivities are related to an α-methylene-γ-lactone moiety and an ester group on carbon C-8 in the sesquiterpene lactone (SL) structure. Two HTSLs, incomptines A (AI) and B (IB) isolated from Decachaeta incompta, were evaluated for their cytotoxic activity on three leukemia cell lines: HL-60, K-562, and REH cells. Both compounds were subjected to a molecular docking study using target proteins associated with cancer such as topoisomerase IIα, topoisomerase IIß, dihydrofolate reductase, methylenetetrahydrofolate dehydrogenase, and Bcl-2-related protein A1. Results show that IA and IB exhibit cytotoxic activity against all cell lines used. The CC50 value of IA was 2-4-fold less than etoposide and methotrexate, two anticancer drugs used as positive controls. The cytotoxic activity of IB was close to that of etoposide and methotrexate. The molecular docking analysis showed that IA and IB have important interaction on all targets used. These findings suggest that IA and IB may serve as scaffolds for the development of new treatments for different types of leukemia.
Subject(s)
Molecular Docking SimulationABSTRACT
The antihyperglycemic activity of ethanolic extract from Salvia polystachya (EESpS) and its products was evaluated using in vivo, ex vivo and in silico assays; additionally, an acute toxicity assay was evaluated. EESpS was classified as a nontoxic class 5 drug. EESpS, ethyl acetate fraction (EtOAcFr), secondary-6-fraction (SeFr6), ursolic acid (UA), and oleanolic acid (OA) reduced the hyperglycemia in DM2 mice. α-glucosidase inhibition was evaluated with oral sucrose and starch tolerance tests (OSuTT and OStTT), an intestinal sucrose hydrolysis (ISH) assay and molecular docking studies using acarbose as control. SGLT1 inhibition was evaluated with oral glucose and galactose tolerance tests (OGTT and OGaTT), an intestinal glucose absorption (IGA) assay and molecular docking studies using canagliflozin as the control. During the carbohydrate tolerance tests, all the treatments reduced the postprandial peak, similar to the control drugs. During the ISH, IC50 values of 739.9 and 726.3 µM for UA and OA, respectively, were calculated. During the IGA, IC50 values of 966.6 and 849.3 for UA, OA respectively, were calculated. Finally, during the molecular docking studies, UA and OA showed ∆G values of -6.41 and -5.48 kcal/mol-1, respectively, on α-glucosidase enzymes. During SGLT1, UA and OA showed ∆G values of -10.55 and -9.65, respectively.
ABSTRACT
Incomptines A (IA) and B (IB) are two sesquiterpene lactones with antiprotozoal, antibacterial, cytotoxic, antitumor, spermicidal, and phytotoxic properties. The antibacterial activity of IA and IB against bacteria causing diarrhoea have been reported; however, no information is available regarding their antibacterial activity on Vibrio cholerae. In this work, both compounds were evaluated for their anti-diarrhoeal potential using the bacterium V. cholerae, sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis on cholera toxin, and a cholera toxin-induced diarrhoea model in male Balb/c mice. In addition, a molecular docking study was carried out to understand the interaction of IA and IB with cholera toxin. In terms of antibacterial activity, IB was three times more active than IA on V. cholerae. In the case of SDS-PAGE analysis and the in silico study, IA was most effective, revealing its potential binding mode at a molecular level. In terms of anti-diarrhoeal activity, IA was 10 times more active than IB and racecadotril, an antisecretory drug used as positive control; the anti-diarrheal activity of IB was also closer than racecadotril. The results obtained from in vitro, in vivo, and computational studies on V. cholerae and cholera toxin support the potential of IA and IB as new anti-diarrhoeal compounds.
ABSTRACT
Incomptine A (IA) is a sesquiterpene lactone isolated from Decachaeta incompta that induces apoptosis, reactive oxygen species production, and a differential protein expression on the U-937 (diffuse histiocytic lymphoma) cell line. In this work, the antitumor potential of IA was investigated on Balb/c mice inoculated with U-937 cells and through the brine shrimp lethality (BSL) test. Furthermore, IA was subjected to molecular docking study using as targets proteins associated with processes of cancer as apoptosis, oxidative stress, and glycolytic metabolism. In addition to determining the potential toxicity of IA in human, its acute toxicity was performed in mice. Results reveals that IA showed high antilymphoma activity and BSL with an EC50 of 2.4 mg/kg and LC50 16.7 µg/mL, respectively. The molecular docking study revealed that IA has strong interaction on all targets used. In the acute oral toxicity, IA had a LD50 of 149 mg/kg. The results showed that the activities of IA including antilymphoma activity, BSL, acute toxicity, and in silico interactions were close to the methotrexate, an anticancer drug used as positive control. These findings suggest that IA may serve as a candidate for the development of a new drug to combat lymphoma.
Subject(s)
Antineoplastic Agents/chemistry , Antineoplastic Agents/pharmacology , Sesquiterpenes/chemistry , Sesquiterpenes/pharmacology , Animals , Cell Line, Tumor , Disease Models, Animal , Dose-Response Relationship, Drug , Humans , Lethal Dose 50 , Ligands , Mice , Molecular Conformation , Molecular Docking Simulation , Molecular Dynamics Simulation , Molecular Structure , Structure-Activity Relationship , Xenograft Model Antitumor AssaysABSTRACT
Sesquiterpene lactones are of pharmaceutical interest due their cytotoxic and antitumor properties, which are commonly found within plants of several genera from the Asteraceae family such as the Decachaeta genus. From Decachaeta incompta four heliangolide, namely incomptines A-D have been isolated. In this study, cytotoxic properties of incomptine A (IA) were evaluated on four lymphoma cancer cell lines: U-937, Farage, SU-DHL-2, and REC-1. The type of cell death induced by IA and its effects on U-937 cells were analyzed based on its capability to induce apoptosis and produce reactive oxygen species (ROS) through flow cytometry with 4',6-diamidino-2-phenylindole staining, dual annexin V/DAPI staining, and dichlorofluorescein 2',7'-diacetate, respectively. A differential protein expression analysis study was carried out by isobaric tags for relative and absolute quantitation (iTRAQ) through UPLC-MS/MS. Results reveal that IA exhibited cytotoxic activity against the cell line U-937 (CC50 of 0.12 ± 0.02 µM) and the incubation of these cells in presence of IA significantly increased apoptotic population and intracellular ROS levels. In the proteomic approach 1548 proteins were differentially expressed, out of which 587 exhibited a fold-change ≥ 1.5 and 961 a fold-change ≤ 0.67. Most of these differentially regulated proteins are involved in apoptosis, oxidative stress, glycolytic metabolism, or cytoskeleton structuration.
Subject(s)
Apoptosis/drug effects , Lymphoma, Non-Hodgkin/metabolism , Proteome/metabolism , Proteomics/methods , Reactive Oxygen Species/metabolism , Sesquiterpenes/pharmacology , Asteraceae/chemistry , Cell Line, Tumor , Cell Survival/drug effects , Chromatography, Liquid/methods , Humans , Lymphoma, Non-Hodgkin/pathology , Protein Interaction Maps/drug effects , Tandem Mass Spectrometry/methods , U937 CellsABSTRACT
Salvia amarissima Ortega was evaluated to determinate its antihyperglycemic and lipid profile properties. Petroleum ether extract of fresh aerial parts of S. amarissima (PEfAPSa) and a secondary fraction (F6Sa) were evaluated to determine their antihyperglycemic activity in streptozo-cin-induced diabetic (STID) mice, in oral tolerance tests of sucrose, starch, and glucose (OSTT, OStTT, and OGTT, respectively), in terms of glycated hemoglobin (HbA1c), triglycerides (TG), and high-density lipoprotein (HDL). In acute assays at doses of 50 mg/kg body weight (b.w.), PEfAPSa and F6Sa showed a reduction in hyperglycemia in STID mice, at the first and fifth hour after of treatment, respectively, and were comparable with acarbose. In the sub-chronic test, PEfAPSa and F6Sa showed a reduction of glycemia since the first week, and the effect was greater than that of the acarbose control group. In relation to HbA1c, the treatments prevented the increase in HbA1c. In the case of TG and HDL, PEfAPSa and F6Sa showed a reduction in TG and an HDL increase from the second week. OSTT and OStTT showed that PEfAPSa and F6Sa significantly lowered the postprandial peak at 1 h after loading but only in sucrose or starch such as acarbose. The results suggest that S. amarissima activity may be mediated by the inhibition of disaccharide hydrolysis, which may be associated with an α-glucosidase inhibitory effect.
Subject(s)
Diabetes Mellitus, Experimental/drug therapy , Diabetes Mellitus, Type 2/drug therapy , Hypoglycemic Agents/chemistry , Salvia/chemistry , Animals , Blood Glucose/metabolism , Camphanes , Diabetes Mellitus, Experimental/blood , Diabetes Mellitus, Experimental/pathology , Diabetes Mellitus, Type 2/blood , Diabetes Mellitus, Type 2/pathology , Drugs, Chinese Herbal/chemistry , Drugs, Chinese Herbal/pharmacology , Glucose/metabolism , Glycated Hemoglobin/metabolism , Humans , Hypoglycemic Agents/pharmacology , Lipid Metabolism/drug effects , Mice , Panax notoginseng , Salvia miltiorrhiza , Triglycerides/bloodABSTRACT
Annona diversifolia Safford and two acyclic terpenoids were evaluated to determine their antihyperglycemic activity as potential α-glucosidase and selective SGLT-1 inhibitiors. Ethanolic extract (EEAd), chloroformic (CHCl3Fr), ethyl acetate (EtOAcFr), aqueous residual (AcRFr), secondary 5 (Fr5) fractions, farnesal (1), and farnesol (2) were evaluated on normoglycemic and streptozocin-induced diabetic mice. EEAd, CHCl3Fr, Fr5, (1) and (2) showed antihyperglycemic activity. The potential as α-glucosidase inhibitors of products was evaluated with oral sucrose and lactose tolerance (OSTT and OLTT, respectively) and intestinal sucrose hydrolysis (ISH) tests; the potential as SGLT-1 inhibitors was evaluated using oral glucose tolerance (OGTT), intestinal glucose absorption (IGA), and urinary glucose excretion (UGE) tests. In OSTT and OLTT, all treatments showed significant activity at two and four hours. In ISH, half maximal effective concentrations (CE50) of 565, 662 and 590 µg/mL, 682 and 802 µM were calculated, respectively. In OGTT, all treatments showed significant activity at two hours. In IGA, CE50 values of 1059, 783 and 539 µg/mL, 1211 and 327 µM were calculated, respectively. In UGE Fr5, (1) and (2) showed significant reduction of the glucose excreted compared with canagliflozin. These results suggest that the antihyperglycemic activity is mediated by α-glucosidase and selective SGLT-1 inhibition.
Subject(s)
Annona/chemistry , Diabetes Mellitus, Experimental/drug therapy , Diabetes Mellitus, Type 2/drug therapy , Hypoglycemic Agents/administration & dosage , Sodium-Glucose Transporter 1/metabolism , Terpenes/administration & dosage , alpha-Glucosidases/metabolism , Administration, Oral , Animals , Blood Glucose/drug effects , Diabetes Mellitus, Experimental/metabolism , Diabetes Mellitus, Type 2/chemically induced , Diabetes Mellitus, Type 2/metabolism , Gene Expression Regulation/drug effects , Glycoside Hydrolase Inhibitors/administration & dosage , Glycoside Hydrolase Inhibitors/chemistry , Glycoside Hydrolase Inhibitors/pharmacology , Hypoglycemic Agents/chemistry , Hypoglycemic Agents/pharmacology , Male , Mice , Molecular Structure , Plant Extracts/chemistry , Plant Extracts/isolation & purification , Rats , Streptozocin , Terpenes/chemistry , Terpenes/pharmacologyABSTRACT
Como importante fonte alimentar humana, o leite e seus derivados sem os devidos cuidados no processamento pode representar um risco à saúde pública. Apesar da legislação brasileira não permitir a comercialização de produtos sem pasteurização, inúmeros produtores alternativamente fabricam queijos e outros derivados a partir de leite cru. Visando melhorar a qualidade e apresentação dos produtos lácteos comercializados por pequenos produtores rurais na cidade de Toledo-PR, foi implantado um projeto com subsídio financeiro do município para aquisição e instalação de pasteurizadores. Com o objetivo de avaliar a qualidade microbiológica dos produtos lácteos produzidos artesanalmente na cidade, foram coletadas amostras de sete produtores de queijo e sete de creme de leite antes e após a pasteurização do leite na propriedade. Análises de Coliformes totais, fecais, E. coli e Salmonella sp. foram realizadas nos produtos em conformidade com a legislação vigente. Quanto à presença de coliformes totais, todas as amostras apresentaram contagem elevada antes da pasteurização e após; esta contagem foi considerada pouco expressiva ou nula. Quanto à presença de coliformes fecais, sete produtores (100%) de queijo e seis produtores (85,71%) de creme de leite apresentaram amostras com contagem acima do permitido pela legislação antes da pasteurização, enquanto que após a pasteurização quatro (57,14%) produtores de queijo e dois (28,57%) de creme de leite se encontraram fora dos padrões exigidos. Foi detectada a presença de E. coli em amostras de cinco produtores (71,42%) de queijo equatro (57,14%) de creme de leite antes da pasteurização e após a pasteurização, confirmando-se em apenas 28,57% e 14,28%, respectivamente. [...] (AU)
As an important human food source, milk and its derivatives may represent a risk to public health. Although brazilian law does not permit the marketing of products without pasteurization, many producers alternatively manufacture cheese and other products from raw milk. To improve the quality and presentation of dairy products marketed by small farmers in the Toledo-PR, a project was implemented financial subsidy for the purchase of pasteurization. Aiming to evaluate the microbiological quality of dairy products produced by craftsmen in the city, samples were collected from seven cheese producers and seven of cream before pasteurization and after pasteurization of milk on the farm. Analysis of total coliforms, fecal, E. coli and Salmonella sp. Products were developed and analyzed in accordance with applicable legislation. For the results, the presence of total coliforms, all samples showed high score for this parameter before and after pasteurization, this count was not considered significant or null. Regarding the presence of fecal coliforms, seven producers (100%) of cheese producers and six (85.71%) of cream samples with results presented above that allowed by law before pasteurization, after pasteurization, while four (57,14%) and two cheese producers (28.57%) cream was found outside the required standards. We detected the presence of E. coli in samples from five producers (71.42%)of cheese and four (57.14%) of cream before pasteurization and after pasteurization, it was confirmed in 28.57% and 14.28% respectively. As the presence of Salmonella sp. Was detected the presence of this organism in samples from four producers (57.14%) and also four cheese cream in the product before pasteurization and after pasteurization was not detected the presence of this microorganism in the samples collected, confirming the efficiency of pasteurization. (AU)
Subject(s)
Cheese/microbiology , Food Microbiology , Food Samples , /microbiology , Food Contamination , Food Handling , ColiformsSubject(s)
Food Microbiology , Food Samples , Cheese/microbiology , Coliforms , Food Contamination , Food Handling , Dairy Products/microbiologyABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: Chiranthodendron pentadactylon Larreat is frequently used in Mexican traditional medicine as well as in Guatemalan for several medicinal purposes, including their use in the control of diarrhea. AIM OF THE STUDY: This work was undertaken to obtain additional information that support the traditional use of Chiranthodendron pentadactylon Larreat, on pharmacological basis using the major antisecretory isolated compound from computational, in vitro and in vivo experiments. MATERIALS AND METHODS: (-)-Epicatechin was isolated from ethyl acetate fraction of the plant crude extract. In vivo toxin (Vibrio cholera or Escherichia coli)-induced intestinal secretion in rat jejunal loops models and sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis on Vibrio cholera toxin were used in experimental studies while the molecular docking technique was used to conduct computational study. RESULTS: The antisecretory activity of epicatechin was tested against Vibrio cholera and Escherichia coli toxins at oral dose 10 mg/kg in the rat model. It exhibited the most potent activity on Vibrio cholera toxin (56.9% of inhibition). In the case of Escherichia coli toxin its effect was moderate (24.1% of inhibition). SDS-PAGE analysis revealed that both (-)-epicatechin and Chiranthodendron pentadactylon extract interacted with the Vibrio cholera toxin at concentration from 80 µg/mL and 300 µg/mL, respectively. Computational molecular docking showed that epicatechin interacted with four amino acid residues (Asn 103, Phe 31, Phe 223 and The 78) in the catalytic site of Vibrio cholera toxin, revealing its potential binding mode at molecular level. CONCLUSION: The results derived from computational, in vitro and in vivo experiments on Vibrio cholera and Escherichia coli toxins confirm the potential of epicatechin as a new antisecretory compound and give additional scientific support to anecdotal use of Chiranthodendron pentadactylon Larreat in Mexican traditional medicine to treat gastrointestinal disorders such as diarrhea.
Subject(s)
Antidiarrheals/pharmacology , Catechin/pharmacology , Diarrhea/drug therapy , Malvaceae , Plant Extracts/pharmacology , Animals , Antidiarrheals/chemistry , Catechin/chemistry , Catechin/isolation & purification , Enterotoxins/administration & dosage , Flowers , Jejunum/metabolism , Male , Medicine, Traditional , Methanol/chemistry , Mexico , Molecular Docking Simulation , Plant Extracts/chemistry , Rats , Rats, Sprague-Dawley , Solvents/chemistryABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: The flowers of Chiranthodendron pentadactylon Larreat. (Sterculiaceae) has been traditionally used as folk medicine in Mexico for the treatment of gastrointestinal disorders such as diarrhea and dysentery. AIM OF THE STUDY: This study aimed to assess the antisecretory activity which supports the therapeutic use of Chiranthodendron pentadactylon and its flavonoids to treat diarrhea. MATERIALS AND METHODS: The methanol extract of Chiranthodendron pentadactylon, subsequent fractions, and flavonoids were evaluated on cholera toxin-induced intestinal secretion in rat jejunal loops model. RESULTS: Three antisecretory flavonoids were isolated by bioassay-guided purification, namely, isoquercitrin 3, (+)-catechin 4 and (-)-epicatechin 5. Among them, epicatechin exhibited the most potent antisecretory activity with ID(50) of 8.3 microM/kg. Its potency was close that of to loperamide (ID(50) 6.1 microM/kg), drug used as control. Isoquercitrin (ID(50) 19.2 microM/kg) and catechin (ID(50) 51.7 microM/kg) showed moderate and weak activity, respectively. CONCLUSION: The results of the present study lend some support to the anecdotal report for the traditional use of the flowers of Chiranthodendron pentadactylon in the control of dysentery.
Subject(s)
Body Fluids , Cholera Toxin/toxicity , Flavonoids/pharmacology , Flowers/chemistry , Intestines/drug effects , Malvaceae/chemistry , Plant Extracts/pharmacology , Animals , Chromatography, Ion Exchange , Female , Intestinal Mucosa/metabolism , Male , Rats , Rats, Sprague-DawleyABSTRACT
A qualidade microbiológica de produtos vegetais constitui atributo essencial para seu desempenho adequado. Neste estudo, foram avaliadas 27 amostras de três espécies vegetais: ginko biloba (Ginkgo biloba), cáscara sagrada (Rhamnus purshiana) e sene (Cassiaangustifolia). O método utilizado foi o de semeadura em profundidade (pour plate) e foram realizadas contagens em placas de Petri com meios seletivos e pesquisa de microrganismos específicos, de acordo com a Organização Mundial da Saúde (OMS). A contaminação bacterianavariou de 0,5 x 101 a 9,0 x 104 unidades formadoras de colônia (UFC)/g, estando todas as amostras em conformidade com a OMS e 5 (18,5 por cento), acima dos limites permitidos pela Farmacopéia Brasileira (FB). A contaminação fúngica variou de 0,5 x 101 a 8,45 x 104 UFC/g, com 6 (22,2 por cento) amostras acima dos limites da OMS e 9 (33,3 por cento) acima dos limites da FB. Foi confirmada a presença de Escherichia coli em 2 (7,4 por cento) amostras e Staphylococcus aureus em 6 (22,2 por cento). Considerando as recomendações da OMS, 22,2 por cento das matérias-primas analisadas foram consideradas insatisfatórias para o consumo e, de acordo com a FB, 37 das amostras apresentaram populações bacterianase fúngicas acima do tolerável, ou continham bactérias não aceitas. Os resultados apontam para a necessidade de controle microbiológico mais rigoroso das drogas vegetais.
The microbiological quality of vegetal products constitutes one of the essential attributes for its adequate performance. In this study, 27 vegetal raw material samples of three species, Ginkgo biloba, Rhamnus purshiana and Cassia angustifolia, were evaluated.The method utilized was pour plate plating and colony counts in Petri dishes with selective medium and investigation of specific microorganism were carried out in accordance with the World Health Organization (WHO). Bacterial contamination varied from 0.5 x 101 to 9.0 x 104 colony-forming units (CFU)/g, all samples in accordance with WHO and 5 (18.5 percent), above the limits allowed by the Brazilian Pharmacopoeia(FB). Fungal contamination varied from 0.5 x 101 to 8.45 x 104 CFU/g, with 6 (22.2 percent) samples above of the limits of WHO and 9 (33.3 percent) above of the limits of the FB. Presence of Escherichia coli in 2 (7.4 percent) and Staphylococcus aureus in 6 (22.2 percent) was confirmed.Considering the recommendations of WHO, 22.2 percent of the analyzed raw materials were considered unsatisfactory for consumption, and, inaccordance with the FB, 37 percent of the samples presented bacterial and fungal populations either above the tolerated or contained unaccepted bacteria. The results point out to the need for more rigorous microbiological control of herbal drugs.
Subject(s)
Phytotherapeutic DrugsABSTRACT
Aqueous and methanolic extracts from 26 medicinal plants used in Mexico to treat gastrointestinal disorders were screened to evaluate their antisecretory activity on cholera toxin-induced intestinal secretion in rat jejunal loops model. Extracts were tested at a dose of 300 mg/kg. From 56 samples tested, both extracts from Chiranthodendron pentadactylon, Hippocratea excelsa and Ocimum basilicum were the most potent with inhibition values ranging from 68.0 to 87.6%. On the other hand, the methanolic extract of Geranium mexicanum (aerial parts) and the aqueous extract of Bocconia frutescens showed the highest activity with inhibition values of 93.4 and 86.0%, respectively. The results obtained in this study give some scientific support to the use of the Mexican medicinal plants employed for the treatment of gastrointestinal disorders such as diarrhea.