【摘要】：微藻的生長條件和環(huán)境特點(diǎn)決定了其胞內(nèi)多糖具有一些有別于其它生物多糖的結(jié)構(gòu)和功能,不同來源的微藻多糖具有不同的生物活性,并且多糖分子中的硫酸基團(tuán)的含量對其生物活性有明顯的影響。如果對多種微藻多糖進(jìn)行硫酸酯化制備,并進(jìn)行相應(yīng)的復(fù)合可篩選出最佳比例復(fù)合多糖,這對今后微藻復(fù)合多糖的開發(fā)及廣泛應(yīng)用有著積極地借鑒意義和促進(jìn)作用。本論文以螺旋藻純多糖(PSP)和小球藻純多糖(CSP)為研究對象,采用濃硫酸法將兩種多糖進(jìn)行了硫酸酯化的制備,并將兩種硫酸酯化多糖按照一定比例配置成15種不同比例的硫酸酯化微藻復(fù)合多糖,通過MTT法分別對這15種不同的硫酸酯化復(fù)合多糖進(jìn)行了體外抗腫瘤的篩選,并采用體外細(xì)胞毒性試驗(yàn)測定了最佳比例多糖的最大無毒濃度。結(jié)果表明,硫酸酯化后的螺旋藻多糖(SPSP)和硫酸酯化后的小球藻多糖(SCSP)的SO42-含量和取代度值分別為19.58%、0.42和19.30%、0.41,傅里葉紅外光譜掃描圖顯示,兩種硫酸酯化多糖不但保存了原多糖的結(jié)構(gòu)而且硫酸基團(tuán)被成功的添加到多糖分子中。15種不同比例的微藻硫酸酯化復(fù)合多糖中SCP12的體外抗腫瘤效果最好,抑制率達(dá)到52.31 %,最大無毒濃度為6.4 mg/mL。采用MTT法測定硫酸酯化復(fù)合多糖SCP12的體外抗腫瘤活性,并以市場購買的藥用PSP作為對照,結(jié)果表明,SCP12對腫瘤細(xì)胞的生長抑制作用明顯,且抑制作用明顯高于市售PSP,在設(shè)定的濃度范圍內(nèi)(0.4-6.4mg/mL)呈現(xiàn)出濃度依賴性,當(dāng)濃度達(dá)到6.4 mg/mL時,SCP12對乳腺癌細(xì)胞(MCF-7)、肝癌細(xì)胞(HepG2)和肺癌細(xì)胞(A549)的抑制率分別為52.31%、39.71%和16.15%;將乳腺癌細(xì)胞轉(zhuǎn)接到小鼠體內(nèi),給予不同劑量治療,結(jié)果顯示,高、中、低(0.8、3.2、6.4mg/mL)三個劑量組的SCP12均可抑制乳腺癌細(xì)胞的生長,其抑制率分別為82.19%、71.32%和28.77%; SCP12體外抗凝血結(jié)果顯示,高、中、低(0.8、3.2、6.4mg/mL)的三個劑量組的SCP12均可改善TT、PT和APTT的值,說明SCP12具有抗凝血作用。硫酸酯化復(fù)合多糖SCP12體外免疫調(diào)節(jié)活性研究表明,在0.4-6.4 mg/mL濃度范圍內(nèi),SCP12能顯著提高小鼠腹腔巨噬細(xì)胞釋放NO的能力、促進(jìn)小鼠脾淋巴細(xì)胞的增殖,其效果明顯高于對照組單糖PSP,但對小鼠腹腔巨噬細(xì)胞吞噬中性紅的能力促進(jìn)作用一般;SCP12體內(nèi)對二硝基氟苯(DNFB)誘導(dǎo)的小鼠遲發(fā)變態(tài)反應(yīng)的結(jié)果顯示,高、中、低劑量組耳腫脹度明顯增加,說明SCP12具有增強(qiáng)因DNFB引起的小鼠遲發(fā)型變態(tài)反應(yīng)。以上結(jié)果表明,硫酸酯化復(fù)合多糖SCP12具有明顯的免疫調(diào)節(jié)作用。
[Abstract]:The growth conditions and environmental characteristics of microalgae determined that the intracellular polysaccharides had some different structures and functions from other biological polysaccharides, and the microalgae polysaccharides from different sources had different biological activities. The content of sulphate groups in polysaccharides has a significant effect on its biological activity. If a variety of microalgae polysaccharides were prepared by sulfate esterification and the corresponding compound could be selected the best proportion of the composite polysaccharides could be selected which would be helpful to the development and wide application of microalgae polysaccharides in the future. In this paper, the pure polysaccharide of Spirulina platensis (PSP) and the pure polysaccharide of Chlorella vulgaris (CSP) were prepared by concentrated sulfuric acid method. Two kinds of sulfated polysaccharides were configured into 15 different proportions of sulfated microalgae polysaccharides according to a certain proportion. The 15 kinds of sulfated polysaccharides were screened by MTT method in vitro. The maximum nontoxic concentration of the optimum proportion of polysaccharides was determined by in vitro cytotoxicity test. The results showed that the SO42- content and degree of substitution of sulfated polysaccharide (SPSP) and chlorella polysaccharide (SCSP) were 19.58 and 19.300.41, respectively. The two sulfated polysaccharides not only preserved the structure of the original polysaccharides, but also the sulfate groups were successfully added to the polysaccharide molecules. 15 different proportions of microalgae sulfated polysaccharides with SCP12 had the best anti-tumor effect in vitro. The inhibition rate was 52.31 and the maximum nontoxic concentration was 6.4 mg / mL. MTT assay was used to determine the antitumor activity of sulfated compound polysaccharide SCP12 in vitro. The results showed that SCP12 had obvious inhibitory effect on the growth of tumor cells. The inhibitory effect was significantly higher than that of PSPs in a concentration dependent manner (0.4-6.4 mg / mL). When the concentration reached 6.4 mg / mL, the inhibition rates of SCP12 on MCF-7, HepG2 and A549 cells were 52.31% and 16.15%, respectively. The growth of breast cancer cells was inhibited by SCP12 in three dose groups (0.8-3.2-6.4mg / mL), the inhibitory rates were 82.191.32% and 28.77mg / mL, respectively. The results of in vitro anticoagulant of SCP12 showed that SCP12 of high, middle and low dose groups (0.83.26.4mg / mL) could improve the values of TTPT and APTT. It indicated that SCP 12 had anticoagulant effect. The immunoregulatory activity of sulfated polysaccharide SCP12 was studied in vitro. The results showed that SCP12 could significantly improve the ability of releasing no from peritoneal macrophages and promote the proliferation of splenic lymphocytes in mice in the concentration range of 0.4-6.4 mg / mL. Its effect was significantly higher than that of control group, but the effect of SCP12 on phagocytosis of neutral red in peritoneal macrophages of mice was higher and higher than that of DNFB induced delayed allergic reaction in mice. The ear swelling in low dose group was significantly increased, indicating that SCP 12 enhanced delayed allergic reaction induced by DNFB in mice. The results showed that the sulfated compound polysaccharide SCP 12 had obvious immunomodulatory effect.
【學(xué)位授予單位】：天津科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R96

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 席波;宋東輝;孫晶;劉鳳路;邸富榮;;十種微藻粗多糖的抑菌作用及海水小球藻粗多糖的抗氧化活性[J];天津科技大學(xué)學(xué)報;2015年05期

2 李廣富;陳偉;范路平;戚慧穎;;茶樹菇多糖酸奶的抗氧化活性與抗衰老研究[J];現(xiàn)代食品科技;2015年10期

3 盧可可;張月巧;袁婭;明建;;硫酸化修飾多糖抗腫瘤活性構(gòu)效關(guān)系及分子機(jī)制研究進(jìn)展[J];食品科學(xué);2014年23期

4 郭龍;楊英來;楊濤;劉子亨;封士蘭;;高硫代度紅芪多糖硫酸酯的制備及體外抗凝血活性研究[J];藥學(xué)學(xué)報;2013年11期

5 胡國元;李超影;陳默;張芊;張哲;;香菇多糖和金針菇多糖的提取及其抑菌活性[J];武漢工程大學(xué)學(xué)報;2013年06期

6 王辰龍;張子奇;王曼;丁之恩;閆曉明;;黑木耳多糖的提取分離及體外抗凝血作用研究[J];食品工業(yè)科技;2013年09期

7 周靜華;左紹遠(yuǎn);;植物多糖生物學(xué)活性研究進(jìn)展[J];大理學(xué)院學(xué)報;2012年06期

8 陳瑋;劉啟順;李曙光;趙小明;彭強(qiáng);高振;胡建恩;杜昱光;;微藻多糖生物活性研究進(jìn)展[J];中國海洋藥物;2012年03期

9 江洪波;毛開云;陳大明;;微藻開發(fā)應(yīng)用的專利分析[J];生物產(chǎn)業(yè)技術(shù);2011年06期

10 黃震;遲秀文;束振華;楊利桃;崔海燕;;海藻多糖對小鼠巨噬細(xì)胞的免疫調(diào)節(jié)作用[J];遼寧中醫(yī)藥大學(xué)學(xué)報;2011年04期

相關(guān)會議論文 前1條

1 宗愛珍;王鳳山;;動物多糖抗腫瘤活性研究進(jìn)展[A];中國藥學(xué)會全國多糖類藥物研究與應(yīng)用研討會論文集[C];2008年

相關(guān)博士學(xué)位論文 前5條

1 余強(qiáng);黑靈芝多糖的免疫調(diào)節(jié)活性及其作用機(jī)制[D];南昌大學(xué);2014年

2 任明;復(fù)合多糖抗輻射和抗腫瘤作用及其機(jī)制研究[D];吉林大學(xué);2014年

3 孫利芹;紫球藻多糖的制備及其生物活性研究[D];大連理工大學(xué);2009年

4 周革非;角叉菜的化學(xué)組成及其抗腫瘤活性的研究[D];中國科學(xué)院研究生院（海洋研究所）;2004年

5 王德培;螺旋藻多糖的研究[D];華南理工大學(xué);1997年

相關(guān)碩士學(xué)位論文 前7條

1 馮雪;紅松松塔多糖膜分離制備工藝及其活性研究[D];青島科技大學(xué);2013年

2 徐紹娜;熟地黃多糖抗突變作用的實(shí)驗(yàn)研究[D];黑龍江中醫(yī)藥大學(xué);2010年

3 穆文靜;鈍頂螺旋藻多糖的提取工藝和其對果蠅抗氧化能力的影響[D];內(nèi)蒙古師范大學(xué);2010年

4 梁進(jìn);茶葉多糖的化學(xué)修飾及體外抗凝血作用研究[D];安徽農(nóng)業(yè)大學(xué);2008年

5 盛建春;蛋白核小球藻（Chlorella pyrenoidosa）多糖的制備和In vitro抗腫瘤活性研究[D];南京農(nóng)業(yè)大學(xué);2007年

6 孫士紅;堿提滸苔多糖降血糖、降血脂生物活性研究[D];東北師范大學(xué);2007年

7 施瑛;蛋白核小球藻多糖制備及其增強(qiáng)免疫功能的研究[D];南京農(nóng)業(yè)大學(xué);2005年

，

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