RESUMO
BACKGROUND: Colon cancer is the most aggressive form of cancers, that causes 0.5 million deaths per year around the globe. Targeting colon cancer by conventional therapeutic options elicits toxicity. Traditional medicines take a lead to alleviate the existing clinical challenges. OBJECTIVE: To investigate antibacterial activity against Helicobacter Pylori and in vitro anti-colon cancer activity by Acacia nilotica extract (ACE) and its active constituent pyrogallol. METHODS: Pyrogallol isolated from A. nilotica by column chromatography and HPLC and structure was elucidated by spectral analysis. Antibacterial activity was done by flow cytometry. Cytotoxicity was measured by MTT assay. Apoptotic morphology and nuclear fragmentation were assessed with AO/ethidium bromide and DAPI staining. DNA fragmentation was done by electrophoresis. Western blot used to analyze the molecular mechanism of apoptosis. Cell cycle arrest was determined using flow cytometry of propidium iodide stained cells. Cell migration was determined by wound healing assay. RESULTS: ACE (20 µg/ml) and pyrogallol (10 µg/ml) treatment reduced the survival of H.pylori at 61% and 62%, respectively. MTT results show that HT-29 cells are more sensitive to pyrogallol with an IC50 value of 35µg/ml compared to ACE. Pyrogallol treated HT-29 cells reached dead state i.e. late apoptotic state with severe nuclear fragmentation. Pyrogallol elicits dose dependent DNA fragmentation in HT-29 cells. Pyrogallol induced apoptosis by simultaneous down-regulation of Bcl-2 and up-regulation of BAX and cytochrome c. Pyrogallol arrested HT-29 cells in S and G2/M phase of cell cycle. Further pyrogallol exhibited marked antimetastatic potential by inhibiting the migration of HT-29 cells dose dependently. CONCLUSION: Both ACE and pyrogallol repressed the growth of H.pylori and as significant anti-colon cancer agent.
Assuntos
Antibacterianos/farmacologia , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Neoplasias do Colo/tratamento farmacológico , Farmacorresistência Bacteriana Múltipla/efeitos dos fármacos , Helicobacter pylori/efeitos dos fármacos , Pirogalol/farmacologia , Antibacterianos/química , Antibacterianos/isolamento & purificação , Antineoplásicos/química , Antineoplásicos/isolamento & purificação , Ciclo Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Neoplasias do Colo/patologia , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Citometria de Fluxo , Células HT29 , Humanos , Testes de Sensibilidade Microbiana , Estrutura Molecular , Pirogalol/química , Pirogalol/isolamento & purificação , Relação Estrutura-Atividade , Cicatrização/efeitos dos fármacosRESUMO
OBJECTIVE: To investigate the prevalence and normal versus abnormal levels of anticardiolipin antibodies (aCL) in a healthy adult population of Omanis and whether a correlation exists between aCL and antinuclear antibodies (ANA) in this Omani population. METHODS: A total of 521 healthy Omani individuals (333 males and 188 females), aged between 17-54-years were investigated for the presence and quantities of anticardiolipin antibodies (immunoglobulin G (IgG)) and IgM isotypes using a conventional enzyme linked immunosorbent assay. Antinuclear antibodies were detected, in this group, using standard indirect immunofluorescence techniques. This study was conducted during the period 2002 through to 2003 at the Immunology Laboratories, College of Medicine and Health Sciences, Sultan Qaboos University, Muscat, Sultanate of Oman. RESULTS: The prevalence of aCL in the healthy Omani population was estimated to be 2.5% for IgG and 3.1% for IgM. The cut off points for IgG and IgM were determined for the whole population as 22.5 IgG phospholipid (GPL) units and 15.7 IgM phospholipid (MPL) units, using the mean plus 5 standard deviations. Using these cut off values, aCL were not detected in the majority of individuals (97%) and in the remaining 3%, the levels were not very high. There was no significant difference between the levels of aCL in either the male or female groups and no significant correlation for the presence of aCL with the age in this studied population. Antinuclear antibodies were detected in 76/521 (14.6%) of the population studied, with some individuals (0.8%) showing titers of 1:640, but there was no association with aCL. CONCLUSION: Although, ANA is present in this healthy Omani population at high frequency and in some individuals at high levels, high levels of aCL do not occur and their presence may be an indicator of autoimmune mediated pathology.