硫酸化修饰对青钱柳多糖抗氧化和抗炎活性的影响
发布时间：2018-10-26 08:09
【摘要】：本论文以青钱柳叶片为原料提取精制青钱柳多糖,命名为CP。对CP进行硫酸化修饰,得青钱柳多糖的硫酸化衍生物S-CP1-4。然后对这两种多糖组分的主要理化性质、单糖组成、分子量、红外光谱、紫外吸收光谱及扫描电镜图谱进行分析;在DPPH自由基体系下,评价其体外抗氧化活性,并建立H2O2诱导的氧化损伤模型,评价其氧化损伤抑制活性;同时研究了硫酸化修饰前后青钱柳多糖对LPS诱导的小鼠炎症的抑制作用。结合理化性质和活性测定的结果,综合评价硫酸化修饰对多糖抗氧化和抗炎活性的影响。主要结果如下:(1)硫酸化修饰可改变青钱柳多糖的基本理化性质、单糖组成和分子量。红外光谱图可证实硫酸根基团被成功的结合到青钱柳多糖的结构中。青钱柳多糖CP是一种含有63.77±1.55%中性糖、11.03±0.17%糖醛酸和8.23±0.78%蛋白质的杂多糖,硫酸化青钱柳多糖S-CP1-4则含有42.41±2.55%的中性糖、13.41±0.69%的糖醛酸和1.95±0.09%的蛋白质。CP中未检出硫酸化基团,S-CP1-4的取代度为0.42。HPGPC色谱表明经硫酸化修饰后,多糖的分子量增加,这是硫酸化基团取代羟基的结果。HPACE结果进一步证明硫酸化修饰对多糖的结构造成一定影响,改变了其单糖组成。同时SEM扫描电镜结果表明,CP和S-CP1-4粉末在扫面电镜中均成片状。(2)硫酸化修饰可提高青钱柳多糖的抗氧化活性。使用H2O2建立RAW264.7氧化损伤模型,结果表明青钱柳多糖CP及其硫酸化衍生物S-CP1-4可以提高小鼠巨噬细胞在H2O2损伤下的存活率,硫酸化修饰可能通过增加超氧化物歧化酶活性和抑制脂质过氧化来减少AW264.7细胞的氧化应激反应。体外自由基清除活性结果表明硫酸化青钱柳多糖具有较高的总还原能力和一定的DPPH自由基清除活性。(3)硫酸化修饰可提高青钱柳多糖的抗炎活性。使用LPS建立小鼠巨噬细胞RAW264.7炎症模型,硫酸化青钱柳多糖可以通过抑制NO释放、减弱吞噬活性、抑制促炎因子的释放起到抑制炎症反应的作用;使用LPS建立小鼠炎症模型,硫酸化青钱柳多糖通过促进抑炎因子IL-10的释放、减少促炎因子TNF-α、IL-6、IL-1β的生成、降低ALT和AST含量、降低脏器指数来发挥抗炎活性,同时S-CP1-4还可以通过促进SOD分泌,抑制MDA生成来减弱炎症导致的氧化损伤。
[Abstract]:In this paper, the leaves of Salix mongolicum were used as raw materials to extract and refine Polysaccharide of Salix mongolicum, named CP.. S-CP1-4, a sulfated derivative of Cyclocycline Polysaccharide, was prepared by sulfation modification of CP. Then the main physicochemical properties, monosaccharide composition, molecular weight, infrared spectrum, UV absorption spectrum and scanning electron microscope (SEM) of the two polysaccharides were analyzed. The antioxidant activity in vitro was evaluated in DPPH free radical system, and the oxidative damage model induced by H2O2 was established to evaluate its oxidative damage inhibition activity. At the same time, the inhibitory effect of Cyclocycline Polysaccharide on LPS induced inflammation in mice was studied before and after sulfated modification. The effects of sulfation modification on the antioxidant and anti-inflammatory activities of polysaccharides were evaluated. The main results were as follows: (1) sulfated modification could change the basic physicochemical properties, monosaccharide composition and molecular weight of the polysaccharide. The IR spectra showed that the sulfate group was successfully bound to the structure of Polysaccharide from Salix Cycariae. CP is a heteropolysaccharide containing 63.77 卤1.55% neutral sugar, 11.03 卤0.17% uronic acid and 8.23 卤0.78% protein, while sulfated S-CP1-4 contains 42.41 卤2.55% neutral sugar. 13.41 卤0.69% uronic acid and 1.95 卤0.09% protein. No sulfated group was detected in CP. The degree of substitution of S-CP1-4 was that the molecular weight of polysaccharide increased after sulfation modification by 0.42.HPGPC chromatography. HPACE results further proved that sulfated modification had a certain effect on the structure of polysaccharide and changed its monosaccharide composition. At the same time, SEM scanning electron microscopy showed that both CP and S-CP1-4 powder were flake in the scanning electron microscope. (2) sulfated modification could improve the antioxidant activity of Polysaccharide of Salix Cycariae. The oxidative damage model of RAW264.7 was established by H2O2. The results showed that CP and its sulfated derivative S-CP1-4 could improve the survival rate of murine macrophages after H2O2 injury. Sulfation modification may reduce the oxidative stress response of AW264.7 cells by increasing the activity of superoxide dismutase and inhibiting lipid peroxidation. The results of free radical scavenging activity in vitro showed that sulfated Willow polysaccharide had high total reduction ability and certain DPPH free radical scavenging activity. (3) sulfated modification could improve the anti-inflammatory activity of Cyclocularia polysaccharide. LPS was used to establish mouse macrophage RAW264.7 inflammatory model. Sulfated Willow Polysaccharide could inhibit inflammation by inhibiting NO release, decreasing phagocytic activity and inhibiting the release of pro-inflammatory factor. LPS was used to establish mouse inflammatory model. Sulfated Willow Polysaccharide reduced the production of TNF- 伪, IL-6,IL-1 尾, ALT and AST by promoting the release of IL-10, reducing the production of TNF- 伪, IL-6,IL-1 尾, and reducing the content of ALT and AST. At the same time, S-CP1-4 can also reduce the oxidative damage induced by inflammation by promoting SOD secretion and inhibiting the production of MDA.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：O629.12
[Abstract]:In this paper, the leaves of Salix mongolicum were used as raw materials to extract and refine Polysaccharide of Salix mongolicum, named CP.. S-CP1-4, a sulfated derivative of Cyclocycline Polysaccharide, was prepared by sulfation modification of CP. Then the main physicochemical properties, monosaccharide composition, molecular weight, infrared spectrum, UV absorption spectrum and scanning electron microscope (SEM) of the two polysaccharides were analyzed. The antioxidant activity in vitro was evaluated in DPPH free radical system, and the oxidative damage model induced by H2O2 was established to evaluate its oxidative damage inhibition activity. At the same time, the inhibitory effect of Cyclocycline Polysaccharide on LPS induced inflammation in mice was studied before and after sulfated modification. The effects of sulfation modification on the antioxidant and anti-inflammatory activities of polysaccharides were evaluated. The main results were as follows: (1) sulfated modification could change the basic physicochemical properties, monosaccharide composition and molecular weight of the polysaccharide. The IR spectra showed that the sulfate group was successfully bound to the structure of Polysaccharide from Salix Cycariae. CP is a heteropolysaccharide containing 63.77 卤1.55% neutral sugar, 11.03 卤0.17% uronic acid and 8.23 卤0.78% protein, while sulfated S-CP1-4 contains 42.41 卤2.55% neutral sugar. 13.41 卤0.69% uronic acid and 1.95 卤0.09% protein. No sulfated group was detected in CP. The degree of substitution of S-CP1-4 was that the molecular weight of polysaccharide increased after sulfation modification by 0.42.HPGPC chromatography. HPACE results further proved that sulfated modification had a certain effect on the structure of polysaccharide and changed its monosaccharide composition. At the same time, SEM scanning electron microscopy showed that both CP and S-CP1-4 powder were flake in the scanning electron microscope. (2) sulfated modification could improve the antioxidant activity of Polysaccharide of Salix Cycariae. The oxidative damage model of RAW264.7 was established by H2O2. The results showed that CP and its sulfated derivative S-CP1-4 could improve the survival rate of murine macrophages after H2O2 injury. Sulfation modification may reduce the oxidative stress response of AW264.7 cells by increasing the activity of superoxide dismutase and inhibiting lipid peroxidation. The results of free radical scavenging activity in vitro showed that sulfated Willow polysaccharide had high total reduction ability and certain DPPH free radical scavenging activity. (3) sulfated modification could improve the anti-inflammatory activity of Cyclocularia polysaccharide. LPS was used to establish mouse macrophage RAW264.7 inflammatory model. Sulfated Willow Polysaccharide could inhibit inflammation by inhibiting NO release, decreasing phagocytic activity and inhibiting the release of pro-inflammatory factor. LPS was used to establish mouse inflammatory model. Sulfated Willow Polysaccharide reduced the production of TNF- 伪, IL-6,IL-1 尾, ALT and AST by promoting the release of IL-10, reducing the production of TNF- 伪, IL-6,IL-1 尾, and reducing the content of ALT and AST. At the same time, S-CP1-4 can also reduce the oxidative damage induced by inflammation by promoting SOD secretion and inhibiting the production of MDA.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：O629.12
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