ABSTRACT
@#To investigate the inhibitory effect of sildenafil on Caco-2 cell proliferation and its anti-inflammatory effect on menadione-induced NCM460 cell inflammation model, MTT assay was used to determine cell proliferation. Intracellular reactive oxygen species(ROS)and nitric oxide(NO)levels were detected by fluorescent probe. Western blot was used to detect the expression of eNOS/ERK/JNK pathway related proteins in Caco-2 cells and correlated inflammatory cytokines in NCM460 cells. The effect of sildenafil on the growth of two probiotics was determined by spectrophotometry. Results showed that sildenafil signi-ficantly inhibited the proliferation of Caco-2 cells and enhanced the expression levels of eNOS, p-eNOS, p-JNK1/2 and p-ERK1/2 proteins in Caco-2 cells; while after adding NG-nitro-L-arginine methyl ester(L-NAME), the expression levels of eNOS, p-eNOS, p-JNK1/2 and p-ERK1/2 proteins were significantly lower than those of the sildenafil group. Compared with the menadione group, sildenafil significantly reduced ROS levels in NCM460 cells and inhibited the expression levels of IL-6, IL-1β, p62, and TNF-α. Moreover, high concentrations of sildenafil had no obvious toxic effects on Lactobacillus casei and Lactobacillus rhamnosus. Thus, the results indicated that sildenafil could effectively inhibit the intestinal inflammatory response without affecting the balance of the intestinal flora, and prevent colorectal cancer by reducing the oxidative stress responses in the intestinal cells.
ABSTRACT
AIM:To investigate the role of microRNA-21(miR-21)in regulation of tight junction(TJ)-asso-ciated protein occludin in human normal colon mucosal epithelial cell line NCM 460, and to analyze related target genes. METHODS:Using miR-21 over-expression lentivirus, the NCM460 cells with miR-21 overexpression were established. The expression level of miR-21 was detected by qPCR.The expression of occludin was determined by Western blot.The target genes of miR-21 were predict by bioinformatic method.According to the scores and the appropriate literature search, a target gene was selected for further study.The miR-21 mimic and inhibitor were transfected into NCM 460 cells,and the expression levels of miR-21 and ROCK1 were measured.Dual-luciferase reporter assay was also performed to verify ROCK1 as the target gene of miR-21.RESULTS:In miR-21-overexpressing NCM460 cells,the protein expression level of occlu-din was upregulated.Bioinformatics and literature analysis predicted that the target gene of miR-21 was ROCK1.The mR-NA and protein levels of ROCK1 were down-regulated in the NCM460 cells transfected with miR-21 mimic.Accordingly, the mRNA and protein levels of ROCK1 were up-regulated in the NCM460 cells transfected with miR-21 inhibitor.Lucifer-ase assay confirmed that miR-21-binding sequence was on the 3'UTR of ROCK1,indicating that ROCK1 is the target gene of miR-21.CONCLUSION: In NCM460 cells, miR-21 up-regulates the expression of occludin, which functions in the maintenance of intestinal epithelial mechanical barrier.ROCK1 is a target gene of miR-21,and is involved in the miR-21 regulation of occludin.
ABSTRACT
O trabalho aborda a aplicação das classificações do Sistema de Informação Estatística em vigência no Brasil na cadeia produtiva agroindustrial, preenchendo assim uma lacuna existente na literatura das ciências agrárias. Devido ao seu destaque no setor, em termos de produção/ comercialização/ exportação, selecionou-se a cadeia produtiva da indústria brasileira óleo-suco-citrícola. O objetivo é identificar o posicionamento de cada classificação na cadeia e as demandas a jusante e a montante para cada atividade e produto. Considerando o sistema estatístico, foi analisada a Classificação Nacional de Atividades Econômicas (CNAE) e a Nomenclatura Comum do Mercosul (NCM). Foram consultadas as informações estatísticas do Instituto Brasileiro de Geografia e Estatística (IBGE) e da Organização das Nações Unidas (ONU), além da específica literatura científica. De modo geral, essas classificações de atividades econômicas e produtos não são mutuamente excludentes, mas podem sobrepor-se umas às outras com relativa facilidade e mínima intervenção. Apesar das muitas segmentações existentes, tanto a cadeia produtiva, quanto as classificações deste trabalho encontram-se corretamente definidas. Os elos da cadeia produtiva da indústria óleo-suco-citrícola brasileira envolvem estas atividades: cultivo, produção, colheita, processamento dos sucos e afins, aproveitamento industrial de subprodutos e resíduos/refugos, packing houses, distribuição, comercialização e consumo. Os produtos são: insumos agrícolas, a laranja (fruta), os sucos, óleos essenciais, águas aromáticas, terpenos, terpenoides, limoneno, perfumes e cosméticos.
This paper addresses the application of the Statistical Information System classifications in force in Brazil in the agro-industrial production chain, thus filling a gap in the literature of agricultural sciences. Due to its prominence in the sector, in terms of production / marketing / exports, it was selected the productive chain of the oil-juice-citrus Brazilian industry. The goal is to identify each classification position in the chain and downstream and upstream demands for each activity and product. Considering the statistical system, it was analyzed the National Classification of Economic Activities (CNAE) and the Mercosul Common Nomenclature (NCM). It was consulted the statistical information of the Brazilian Institute of Geography and Statistics (IBGE) and the United Nations (UN), besides the specific scientific literature. In general, these classifications of economic activities and products are not mutually exclusionary but can overlap each other with relative ease and minimal intervention. Despite the many existing segmentation, both the supply chain and the classifications from this work are set correctly. The productive chain of the oil-juice-citrus Brazilian industry involves these activities: cultivation, production, harvesting, processing of juices and related, industrial utilization of by-products and waste / scrap, packing houses, distribution, marketing and consumption. The products are: agricultural inputs, orange (fruit), juices, essential oils, aromatic waters, terpenes, terpenoids, limonene, fragrance and cosmetics.