| Moringa (Moringa Oleifera), also known as drumstick tree, belonging to woody plants, not only can be eaten directly, after processing can also be made into health care products, cosmetics, has also been studied many experimental personnel as test materials. It is rich in nutrients, mainly calcium, iron, polysaccharide, flavonoids and other active ingredients, which belongs to one of the Moringa polysaccharide. The study found that the polysaccharide consists of animals, plants and microorganisms of three kinds of polysaccharides, biologically active antiviral, antitumor, and has low toxicity, wide sources, easy preparation and other advantages, can be used as an ideal natural medicine and functional food development and utilization. At present, many domestic and foreign literature on the activity of polysaccharide, such as yam polysaccharide anti parvovirus Scallop in Shell polysaccharide, anti hepatitis B.virus, but in Moringa and polysaccharide are rarely reported, but only in the extraction and separation of M.oleifera composition, also did not carry out research on anti virus activity of M.oleifera aspects, no more the study of anti hepatitis C virus.. In this paper, M.oleifera as raw materials, systematically studied the extraction and purification of Moringa oleifera polysaccharides, separation, and anti HCV with its WST-1 method and fluorescence quantitative PCR method for biological activity and mode of action were discussed.1.Moringa, extraction and purification of polysaccharide. The hot water extraction method, the extraction process of extraction:solid-liquid ratio 1:10, extraction temperature of 80℃, extraction time 90min, extraction times 3, and use the DEAE Sepharose Fast Flow fast flow fiber column on the concentration of 0.1mol/L-lmol/L dissolved in NaCl of Moringa oleifera polysaccharides were isolated and purified, were investigated by Sevage, three chloroacetic acid and enzyme hydrolysis in addition to the three kinds of methods of protein purification, optimization of the extraction process, and the content of polysaccharide was determined by phenol sulfuric acid method. Studies show that:Sevage method to remove protein from Moringa polysaccharide, the appearance of deep yellow, massive, easily soluble in hot water, the content of polysaccharide was 29.5%; removal of protein polysaccharide obtained by three trichloroacetic acid method, the appearance of yellow white, powder, easily soluble in hot water, the content of polysaccharide is 31.6%; enzymatic hydrolysis of the polysaccharide, the appearance of light yellow, cottony, easily soluble in cold water, the polysaccharide content was 39.5%. The above three kinds of polysaccharides obtained by the method are easy to get damp, need to pay attention to dry preservation. Sevage method to remove protein reagent waste is serious, and the polysaccharide loss together with protein, resulting in a loss of polysaccharide; three trichloroacetic acid method to remove protein polysaccharide prone to degradation; enzymatic extraction of polysaccharide polysaccharide assisted hot water, not easy to be degraded, and can maintain higher activity, but also can significantly improve the content sugar, is an ideal method for extracting polysaccharide.2.Study on the antiviral activity of Moringa polysaccharide. The method of cell culture in vitro activity of Polysaccharide from Moringa oleifera anti HCV were studied. Four water soluble tetrazolium (WST-1) method for determination of Polysaccharide on human hepatocellular carcinoma (Huh7.5.1) cell toxicity, using fluorescence quantitative PCR method to investigate the Moringa antiviral activity, and using three different inoculation methods, namely, add sugar after virus before adding virus added sugar at the same time, virus polysaccharide, preliminary study on the effect of Moringa polysaccharide antiviral. The results showed that the concentration in the range of 7.8μg/mL-125μg/mL in the WST-1 trial, Moringa oleifera polysaccharides on Huh7.5.1 cell toxicity, the safe concentration range. Determination of Moringa polysaccharide can get antiviral activity against HCV by fluorescent PCR method, and the effect of virus and add sugar after first is better than the other two methods, and the optimal concentration of polysaccharide was 62.5 μg/mL,there was a concentration dependent, adding polysaccharide concentration increases, anti the activity of HCV is better, suggesting that in the clinical application of Moringa polysaccharide can be used to prevent. |
