【摘要】：本論文以青錢柳葉片為原料提取精制青錢柳多糖,命名為CP。對(duì)CP進(jìn)行硫酸化修飾,得青錢柳多糖的硫酸化衍生物S-CP1-4。然后對(duì)這兩種多糖組分的主要理化性質(zhì)、單糖組成、分子量、紅外光譜、紫外吸收光譜及掃描電鏡圖譜進(jìn)行分析;在DPPH自由基體系下,評(píng)價(jià)其體外抗氧化活性,并建立H2O2誘導(dǎo)的氧化損傷模型,評(píng)價(jià)其氧化損傷抑制活性;同時(shí)研究了硫酸化修飾前后青錢柳多糖對(duì)LPS誘導(dǎo)的小鼠炎癥的抑制作用。結(jié)合理化性質(zhì)和活性測定的結(jié)果,綜合評(píng)價(jià)硫酸化修飾對(duì)多糖抗氧化和抗炎活性的影響。主要結(jié)果如下:(1)硫酸化修飾可改變青錢柳多糖的基本理化性質(zhì)、單糖組成和分子量。紅外光譜圖可證實(shí)硫酸根基團(tuán)被成功的結(jié)合到青錢柳多糖的結(jié)構(gòu)中。青錢柳多糖CP是一種含有63.77±1.55%中性糖、11.03±0.17%糖醛酸和8.23±0.78%蛋白質(zhì)的雜多糖,硫酸化青錢柳多糖S-CP1-4則含有42.41±2.55%的中性糖、13.41±0.69%的糖醛酸和1.95±0.09%的蛋白質(zhì)。CP中未檢出硫酸化基團(tuán),S-CP1-4的取代度為0.42。HPGPC色譜表明經(jīng)硫酸化修飾后,多糖的分子量增加,這是硫酸化基團(tuán)取代羥基的結(jié)果。HPACE結(jié)果進(jìn)一步證明硫酸化修飾對(duì)多糖的結(jié)構(gòu)造成一定影響,改變了其單糖組成。同時(shí)SEM掃描電鏡結(jié)果表明,CP和S-CP1-4粉末在掃面電鏡中均成片狀。(2)硫酸化修飾可提高青錢柳多糖的抗氧化活性。使用H2O2建立RAW264.7氧化損傷模型,結(jié)果表明青錢柳多糖CP及其硫酸化衍生物S-CP1-4可以提高小鼠巨噬細(xì)胞在H2O2損傷下的存活率,硫酸化修飾可能通過增加超氧化物歧化酶活性和抑制脂質(zhì)過氧化來減少AW264.7細(xì)胞的氧化應(yīng)激反應(yīng)。體外自由基清除活性結(jié)果表明硫酸化青錢柳多糖具有較高的總還原能力和一定的DPPH自由基清除活性。(3)硫酸化修飾可提高青錢柳多糖的抗炎活性。使用LPS建立小鼠巨噬細(xì)胞RAW264.7炎癥模型,硫酸化青錢柳多糖可以通過抑制NO釋放、減弱吞噬活性、抑制促炎因子的釋放起到抑制炎癥反應(yīng)的作用;使用LPS建立小鼠炎癥模型,硫酸化青錢柳多糖通過促進(jìn)抑炎因子IL-10的釋放、減少促炎因子TNF-α、IL-6、IL-1β的生成、降低ALT和AST含量、降低臟器指數(shù)來發(fā)揮抗炎活性,同時(shí)S-CP1-4還可以通過促進(jìn)SOD分泌,抑制MDA生成來減弱炎癥導(dǎo)致的氧化損傷。
[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 鹵1.55% neutral sugar, 11.03 鹵0.17% uronic acid and 8.23 鹵0.78% protein, while sulfated S-CP1-4 contains 42.41 鹵2.55% neutral sugar. 13.41 鹵0.69% uronic acid and 1.95 鹵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- 偽, IL-6,IL-1 尾, ALT and AST by promoting the release of IL-10, reducing the production of TNF- 偽, 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.
【學(xué)位授予單位】：南昌大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：O629.12

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本文編號(hào)：2295103