ABSTRACT
The purple yam, Dioscorea alata L., is an important source of starch, vitamins and polyphenols. Five different pigments from the purple tubers of this plant were separated by high-performance liquid chromatography-mass spectrometry, and the anthocyanin fraction (DACN) was collected. The anti-inflammatory effects of DACNs were investigated at different concentrations and compared with the standard colitis treatment, 5-aminosalicylic acid, in a trinitrobenzenesulfonic acid (TNBS)-induced colitis mouse model. Macro- and microscopic parameters including body weight change, disease activity index (DAI) and intestinal histology were used for the determination of the anti-inflammatory effects of DACNs. The gene expression levels of tight junction-related proteins in the intestine, myeloperoxidase activity, inducible nitric oxide synthase activity in colonic tissues and pro-inflammatory cytokine production in serum were also measured to elucidate the mechanism of DACN action. Eighty micrograms of DACNs per kilogram of body weight produced potent anti-inflammatory effects in the mouse model of inflammatory bowel disease (IBD), as shown by the DAI (2.78 ± 0.38 vs. 0.44 ± 0.51). Therefore, DACNs may be applied as a potential food supplement in IBD therapy.
Subject(s)
Anthocyanins/administration & dosage , Anti-Inflammatory Agents/administration & dosage , Colitis/drug therapy , Dioscorea/chemistry , Plant Extracts/administration & dosage , Animals , Colitis/chemically induced , Colitis/genetics , Colitis/immunology , Cytokines/genetics , Cytokines/immunology , Disease Models, Animal , Humans , Male , Mice , Mice, Inbred C57BL , Trinitrobenzenesulfonic Acid/adverse effectsABSTRACT
A plasmid was constructed for quantification of genetically modified (GM) cottonseed meal in the gene-specific level. The Cry1Ab/c gene was connected with the Sad1 gene by fusion PCR. The fusion gene was cloned into the pMD®19-T Simple Vector. The plasmid DNA was then digested with a restriction endonuclease SmaI to reduce the characteristic differences between the plasmid DNA and genomic DNA. For a rough quantitative analysis of GM cotton meal contents, a rapid method for measurement of the copy numbers of the transgenic Cry and cotton endogenous Sad1 gene using a real-time PCR system with the plasmid DNA as a calibrator was established. The inter-run and intra-run coefficients of variation were less than 1.48% and 2.36%, respectively. The limits of detection and quantitation of the Cry and Sad1 genes were 9 and 91 copies of pMDCS, respectively. These results prove that the standard plasmid represents a valuable alternative to genomic DNA as a certified reference material for the quantification of GM cotton and is a useful tool to establish a feasible identification management for GM cottonseed meal content in the feed industry.