【摘要】：硫酸軟骨素有著重要的生物學(xué)功能,但鱘魚硫酸軟骨素免疫功能相關(guān)研究報(bào)道較少。另外,硫酸軟骨素已被證實(shí)有著較好的抗腫瘤生物活性,但其機(jī)理還不清楚。故本論文通過對(duì)清潔級(jí)KM小鼠灌胃給藥不同劑量鱘魚硫酸軟骨素2周后,采用遲發(fā)超敏反應(yīng)、二甲苯誘發(fā)耳過敏性反應(yīng)、蛋清致足墊炎、石蠟組織切片實(shí)驗(yàn)方法觀察鱘魚硫酸軟骨素抗炎抗過敏活性和免疫功能活性;以及不同劑量鱘魚硫酸軟骨素(0.03mg/kg,0.3mg/kg和3mg/kg)對(duì)肝癌H_(22)荷瘤小鼠的影響進(jìn)行研究。為全面認(rèn)識(shí)鱘魚硫酸軟骨素的免疫功能以及抗腫瘤生物活性研究提供基礎(chǔ)資料。通過研究得到如下結(jié)果:1、鱘魚硫酸軟骨素可顯著增加小鼠胸腺和脾臟重量指數(shù)(P0.05),增加外周血白細(xì)胞數(shù)量(P0.01),顯著提高遲發(fā)性超敏反應(yīng)強(qiáng)度(P0.01),提高特異性抗體產(chǎn)生能力(P0.05),并顯著降低二甲苯致敏的耳腫脹度及蛋清致足墊炎腫脹度(P0.05)。2、鱘魚硫酸軟骨素能顯著促進(jìn)免疫器官中肥大細(xì)胞數(shù)量的增長。其中,600mg/kg劑量下小鼠胸腺指數(shù)顯著(P0.05)增加,胸腺中肥大細(xì)胞數(shù)量極顯著(P0.01)增加,60mg/kg劑量下小鼠脾臟重量的增加最為明顯(P0.01),6mg/kg劑量下小鼠脾臟中肥大細(xì)胞數(shù)量的增加最為明顯(P0.01)。3、采用四噻唑藍(lán)(MTT)法,用酶標(biāo)儀在波長490nm處測定OD值,觀察鱘魚硫酸軟骨素對(duì)肝癌細(xì)胞體外培養(yǎng)生長的影響,結(jié)果表明:鱘魚硫酸軟骨素對(duì)H_(22)7402和H_(22)7721細(xì)胞的體外培養(yǎng)無明顯的抑制作用,對(duì)H_(22)HepG2細(xì)胞的體外培養(yǎng)有抑制作用,在鱘魚硫酸軟骨素的最終濃度為1000ug/ml和125ug/ml時(shí),對(duì)腫瘤細(xì)胞的抑制效果較好。4、對(duì)清潔級(jí)KM小鼠連續(xù)灌胃給藥不同劑量鱘魚硫酸軟骨素10天后,腋窩皮下接種H_(22)腫瘤細(xì)胞株,觀察鱘魚硫酸軟骨素預(yù)防性給藥后對(duì)小鼠體重、瘤體大小、重量和抑瘤率。結(jié)果表明:鱘魚硫酸軟骨素能促進(jìn)肝癌H_(22)荷瘤小鼠體重增加,并能顯著抑制腫瘤的生長。其中,在60mg/kg劑量下小鼠體重顯著增加(P0.05),腫瘤體積顯著減小(P0.05)。5、對(duì)清潔級(jí)KM小鼠腋窩皮下接種H_(22)腫瘤細(xì)胞株24h后連續(xù)灌胃給藥不同劑量鱘魚硫酸軟骨素1周,觀察各組小鼠體重、瘤體大小、重量,并用外科手術(shù)取出腫瘤稱重,計(jì)算抑瘤率,觀察鱘魚硫酸軟骨素對(duì)肝癌H_(22)荷瘤小鼠體重、腫瘤體積重量和抑瘤率的影響。結(jié)果表明:鱘魚硫酸軟骨素能促進(jìn)肝癌H_(22)荷瘤小鼠體重增加,并能顯著抑制腫瘤的生長。其中,60mg/kg劑量下肝癌H_(22)荷瘤小鼠體重顯著增加(P0.05),600mg/kg劑量下肝癌H_(22)荷瘤小鼠腫瘤體積顯著減小((P0.05),60mg/kg劑量下肝癌H_(22)荷瘤小鼠腫瘤重量顯著減少(P0.05)。(1153)
[Abstract]:Chondroitin sulfate has important biological function, but there are few reports on immune function of chondroitin sulfate in Sturgeon. In addition, chondroitin sulfate has been proved to have good anti-tumor biological activity, but its mechanism is not clear. Therefore, in this paper, two weeks after different doses of chondroitin sulfate were given to clean km mice, delayed hypersensitivity reaction was used, xylene induced ear hypersensitivity and egg white induced foot pad inflammation. The effects of different doses of chondroitin sulfate (0.03 mg / kg, 0.3 mg / kg and 3mg/kg) on H22 tumor bearing mice were studied. To provide basic information for the study of immune function and anti-tumor activity of chondroitin sulfate in Sturgeon. The results showed that chondroitin sulfate significantly increased the weight index of thymus and spleen (P0.05), the number of white blood cells in peripheral blood (P0.01), the intensity of delayed hypersensitivity reaction (P0.01) and the ability of producing specific antibodies. The results showed that chondroitin sulfate significantly increased the number of mast cells in immune organs and significantly decreased the swelling degree of xylene sensitized ears and egg white induced foot pad swelling (P0.05). The thymus index of mice increased significantly (P0.05) at the dose of 600 mg / kg. The number of mast cells in thymus was significantly increased (P0.01). The increase of spleen weight was most obvious (P0.01) at the dose of 60 mg / kg (P0.01) 6 mg / kg (P0.01). The number of mast cells in spleen was increased most obviously (P0.01). 3. The method of tetrathiazolyl blue (MTT) was used. The OD value of Sturgeon chondroitin sulfate was measured at wavelength 490nm, and the effect of chondroitin sulfate on the growth of hepatoma cells was observed. The results showed that chondroitin sulfate had no obvious inhibitory effect on the culture of H _ (22) 7402 and H _ (22) 7721 cells in vitro. When the final concentration of chondroitin sulfate in Sturgeon was 1000ug/ml and 125ug/ml, the inhibitory effect of Chondroitin sulfate on tumor cells was better. 4. After 10 days of continuous oral administration of chondroitin Sturgeon sulfate in clean grade km mice, different doses of chondroitin Sturgeon sulfate were obtained. H22 tumor cell line was inoculated subcutaneously under axilla. The weight, tumor size, weight and tumor inhibition rate of Sturgeon chondroitin sulfate were observed after prophylactic administration of chondroitin sulfate. The results showed that chondroitin sulfate from Sturgeon could promote the weight gain and inhibit the growth of hepatoma H22 mice. The weight of mice increased significantly at the dose of 60mg/kg (P0.05), and the volume of tumor decreased significantly (P0.05). The weight of each group was observed by intragastric administration of Chondroitin Sturgeon sulfate for 24 hours after subcutaneous inoculation of H22 tumor cell line in clean grade km mice. The tumor size and weight were taken out and the tumor inhibition rate was calculated. The effects of chondroitin sulfate on the body weight tumor volume and tumor inhibition rate of H22 bearing hepatoma mice were observed. The results showed that chondroitin sulfate from Sturgeon could promote the weight gain and inhibit the growth of hepatoma H22 mice. At the dose of 60mg / kg, the weight of hepatoma H22 bearing mice increased significantly (P0.05), and the tumor volume of H22 tumor bearing mice decreased significantly (P0.05). (P0.05) the tumor weight of H22 tumor bearing mice decreased significantly (P0.05) (P0.05) at the dose of 60mg / kg (P0.05).
【學(xué)位授予單位】：集美大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R915

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