【摘要】：水提取法提取繡球菌粗多糖,進(jìn)行脫色和脫蛋白處理,分離純化出多糖組分,測(cè)其單糖組成與紅外光譜分析。結(jié)果表明D301T離子交換樹(shù)脂脫色率達(dá)到了55.67%,是本試驗(yàn)中繡球菌多糖脫色的最佳樹(shù)脂。從繡球菌多糖中主要分離出兩個(gè)組分,蒸餾水洗脫的組分(SCPl)和0.1%NaCl洗脫的組分(SCP2),SCP1與SCP2單糖組成都主要為甘露糖(Man)、葡萄糖(Glu)、半乳糖(Gla)和巖藻糖(Fuc),其摩爾比分別為0.043：0.06：1.75：0.386和0.0554：1.656：0.1249：0.0201。紅外光譜分析得到其都具有多糖特征結(jié)構(gòu)。采用體外抗氧化試驗(yàn)與建立衰老模型、灌胃不同濃度繡球菌多糖(SCP)進(jìn)行體內(nèi)抗氧化試驗(yàn)。結(jié)果表明在體外試驗(yàn)中繡球菌多糖(SCP)及其組分SCP1和SCP2對(duì)羥自由基、ABTS和DPPH均有很好的清除效果,同時(shí)具有很好的還原力,且SCP2作用最為突出。體內(nèi)試驗(yàn)中,繡球菌多糖能夠降低小鼠體內(nèi)的MDA同時(shí)提高SOD與CAT含量,抗氧化效果較好。采用MTT法、倒置顯微鏡觀察法和熒光染色法研究繡球菌多糖不同組分抑制人肝癌HepG-2細(xì)胞增殖作用。結(jié)果表明繡球菌多糖組分對(duì)人肝癌HepG-2細(xì)胞有較好的抑制作用,SCP2的抑制作用最好,其作用24h與48h的抑制率分別為48.12%和56.45%,倒置顯微鏡法與熒光染色法都能觀察到繡球菌多糖具有使人肝癌HepG-2細(xì)胞凋亡的作用。建造小鼠腫瘤模型,灌胃不同濃度繡球菌多糖(SCP),測(cè)定小鼠腫瘤大小、脾臟與胸腺指數(shù)和血清中TNF-a與TNF-P含量,對(duì)腫瘤組織進(jìn)行石蠟切片觀察,同時(shí)對(duì)小鼠進(jìn)行生存試驗(yàn)。結(jié)果表明繡球菌多糖對(duì)S180實(shí)體瘤小鼠的腫瘤有明顯的抑制作用,繡球菌多糖高劑量組最高抑制率達(dá)到50%；對(duì)小鼠脾臟與胸腺指數(shù)均有提高作用,且能增加小鼠體內(nèi)TNF-a與TNF-p的含量；石蠟切片觀察腫瘤組織發(fā)現(xiàn)其呈現(xiàn)凋亡病理學(xué)特征；繡球菌多糖對(duì)小鼠生存壽命有一定延長(zhǎng)作用,高劑量組對(duì)小鼠壽命延長(zhǎng)32.6%。本研究表明,繡球菌多糖具有較高的生物活性,其中純化的多糖SCP1是以半乳糖為主的多糖。SCP2是以葡萄糖為主的多糖。以葡萄糖為主的SCP2組分生物活性最高,且具有良好抗氧化功效,在體外對(duì)人肝癌HepG-2細(xì)胞的抑制作用也最好。繡球菌多糖在體內(nèi)能抑制S180實(shí)體瘤小鼠的腫瘤和延長(zhǎng)腫瘤小鼠生存壽命的抗腫瘤與作用抗氧化功效,是很值得重視和開(kāi)發(fā)的藥用真菌。
[Abstract]:The polysaccharide was extracted by water extraction, decolorized and deproteinized. The polysaccharide components were separated and purified. The monosaccharide composition of the polysaccharide was analyzed by infrared spectroscopy. The results showed that the decolorization rate of D301T ion exchange resin was 55.67, which was the best resin for decolorizing hydrangea polysaccharide. Two main components were isolated from the polysaccharides of Hydrangea, distilled water eluting components (SCPl) and 0.1%NaCl elution components (SCP2), and SCP1 and SCP2 monosaccharide groups in Chengdu were mainly mannose (Man), glucose (Glu),. The molar ratios of galactose (Gla) and fucose (Fuc), are 0.043: 0.06: 1.75: 0.386 and 0.0554: 1.656: 0.1249: 0.0201, respectively. Infrared spectrum analysis showed that all of them had the characteristic structure of polysaccharides. Antioxidation test in vitro and establishment of aging model were carried out in vivo with different concentrations of Hydrangea polysaccharide (SCP). The results showed that (SCP), its components SCP1 and SCP2 had good scavenging effect on hydroxyl radical, ABTS and DPPH in vitro, and had good reducing power, and SCP2 had the most prominent effect. In vivo experiment, Hydrangea polysaccharide could decrease MDA and increase the contents of SOD and CAT in mice, and the antioxidant effect was better. The effects of different components of hydrangea polysaccharides on the proliferation of human hepatoma HepG-2 cells were studied by MTT, inverted microscope and fluorescence staining. The results showed that the polysaccharides of Hydrangea had a better inhibitory effect on human hepatoma HepG-2 cells, and SCP2 had the best inhibitory effect. The inhibitory rates of SCP2 at 24h and 48h were 48.12% and 56.45%, respectively. It was observed by inverted microscope and fluorescence staining that Hydrangea polysaccharide could induce apoptosis of human hepatoma HepG-2 cells. The tumor model was established in mice. The tumor size, spleen and thymus index, TNF-a and TNF-P content in serum were measured by (SCP), with different concentrations of hydrangea polysaccharide. The tumor tissue was observed by paraffin section. At the same time, the survival test of mice was carried out. The results showed that Hydrangea polysaccharide had obvious inhibitory effect on tumor of S180 solid tumor mice, and the highest inhibitory rate of Hydrangea polysaccharide group was 50%. The indexes of spleen and thymus were increased, and the contents of TNF-a and TNF-p in mice were increased, and the pathological characteristics of apoptosis were found by paraffin section. Hydrangea polysaccharide could prolong the life span of mice to a certain extent, and the life span of mice in high dose group was increased by 32.6%. The results showed that the polysaccharides of Hydrangea had high bioactivity. The purified polysaccharide SCP1 was galactose and SCP2 was glucose. The SCP2 component with glucose as the main component has the highest bioactivity and has good antioxidant activity. It also has the best inhibitory effect on human hepatoma HepG-2 cells in vitro. Hydrangea polysaccharide can inhibit the tumor of S180 solid tumor mice in vivo and prolong the survival life of tumor mice. It is worthy of attention and development of medicinal fungi.
【學(xué)位授予單位】：福建師范大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R915

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