本文選題：酵母多糖 + 抗癌　； 參考：《吉林農(nóng)業(yè)大學(xué)》2015年博士論文

【摘要】：食藥用菌是人類重要的食品、藥品資源。研究食藥用菌的活性物質(zhì)機理、作用和前景,對食藥用菌的深度開發(fā)、推廣,從而造福人類具有深遠的意義。酵母,作為重要的真菌之一,已有數(shù)千年應(yīng)用歷史。現(xiàn)代生物學(xué)的發(fā)展對酵母有了更深入和廣泛的研究和應(yīng)用。酵母細胞壁富含多糖,成本低廉、供應(yīng)充足,是應(yīng)用廣泛的真菌多糖。酵母多糖主要成分是葡聚糖和甘露寡糖,具有多種生物活性。研究利用葡聚糖與甘露寡糖可用稀酸水解的化學(xué)性質(zhì)這一要點,成功建立了同時檢測葡聚糖與甘露寡糖含量的HPLC方法。本文利用腸道出血綜合癥(一種危害高產(chǎn)奶牛的毀滅性疾病)和煙曲霉(Aspergillus fumigatus)研究酵母多糖對免疫低下個體的免疫力恢復(fù)能力,以及找到相關(guān)的免疫標(biāo)記,建立相關(guān)檢測體系。這一部分建立了一種實時定量煙曲霉菌PCR檢測方法,以定量分析腸道出血綜合癥中奶牛血液及組織樣品內(nèi)的煙曲霉DNA。第一項研究表明,腸道出血綜合癥的發(fā)生與煙曲霉之間存在正相關(guān)。由于煙曲霉具有攻擊感染免疫損傷(immunocomprisied)個體的特性,因此,這部分研究進行了一項以綿羊為對象的實驗,以驗證上述假設(shè),并且檢測了真菌多糖在調(diào)節(jié)先天、后天免疫功能的能力。該研究確認了地塞米松注射處理生長期的綿羊可以建立免疫損傷模型,每日注射地塞米松降低了L-選擇素與IL-1β的濃度,而通過添加酵母多糖可以恢復(fù)免疫抑制綿羊的L-選擇素與IL-1β的基因表達水平。酵母多糖通過提升L-選擇素與IL-1β的表達,加強了嗜中性粒細胞的功能,并且抑制凋亡信號,強化嗜中性粒細胞準(zhǔn)確到達靶位點的功能,使其有效進入感染部位并殺滅病原體。本文研究了酵母真菌多糖免疫調(diào)節(jié)功能及其抗腫瘤作用機制。實驗中,我們研究了酵母多糖對正常小鼠、免疫抑制小鼠免疫功能的調(diào)節(jié)作用,進一步利用H22荷瘤小鼠模型研究酵母多糖通過提高免疫系統(tǒng)達到抗癌效果的作用機理。通過統(tǒng)計分析荷瘤小鼠腫瘤重量、免疫器官指數(shù)和相關(guān)細胞因子含量的變化,探討其可能的抗腫瘤作用機制。實驗結(jié)果顯示,酵母真菌多糖具有抑瘤作用,并使小鼠體重增加,提高免疫器官指數(shù)。環(huán)磷酰胺(CTX)是一種烷化劑類抗腫瘤藥,對機體免疫力起著一定程度的抑制作用,雖能有效抑制腫瘤,但同時也有很強的副作用,造成CTX組小鼠的免疫器官呈現(xiàn)不同程度的萎縮、相關(guān)免疫指標(biāo)降低,但酵母多糖對因其產(chǎn)生的副作用有明顯的調(diào)節(jié)作用。通過小白鼠的飼喂實驗,酵母多糖對小鼠脾臟和胸腺等臟器官有明顯的促進作用。真菌多糖可顯著提高一系列免疫標(biāo)記,包括TNF-α、IFN-γ、CRP及IL-6等。這些基因在腸道細胞中的表達(P0.05)。運用ELISA檢測了鼠血清的TNF-α,IFN-γ,CRP和IL-6的濃度,統(tǒng)計結(jié)果表明這些細胞因子濃度在實驗組有顯著提高(P0.05)。說明酵母多糖通過促進細胞因子的基因表達來促進機體的免疫調(diào)節(jié)功能。本研究專門純化了血樣中的嗜中性粒細胞,進而研究真菌多糖對其TNF-α、IFN-γ、CRP及IL-6表達的影響,結(jié)果表明,酵母多糖顯著提高這些細胞因子的表達(P0.05),并且提高了L-selectin和IL-8R在腸道和嗜中性粒細胞的表達(P0.05),增強了嗜中性粒細胞的吞噬能力。由于所選用的標(biāo)記的特殊性,表明在先天性免疫和后天性免疫,包括體液和細胞免疫中,酵母多糖通過減少免疫抑制和增強功能性基因的表達來提高免疫細胞的抗癌和抗病能力的作用,為了探討酵母多糖在體外影響腫瘤細胞活性的抗腫瘤作用,通過對乳腺癌MCF-7細胞和卵巢癌SKOV3細胞的抑制率進行了研究,結(jié)果表明酵母多糖對兩株癌細胞的增殖具有一定的抑制作用。本研究進行了肉雞和生豬動物實驗,以驗證酵母多糖對免疫功能的調(diào)節(jié)作用在養(yǎng)殖動物的應(yīng)用效果。煙曲霉的侵染導(dǎo)致肉雞出現(xiàn)高死亡率高、生長緩慢等現(xiàn)象。煙曲霉的侵染亦導(dǎo)致母豬腹瀉、死胎和流產(chǎn),新生仔豬拉痢并于數(shù)天內(nèi)死亡。而酵母多糖提高了仔豬成活率和斷奶體重,降低了母豬繁殖性疾病,增加了產(chǎn)仔數(shù),提高發(fā)情率。兩組實驗中,酵母多糖(葡聚糖和甘露寡糖)的使用緩解了煙曲霉對動物的應(yīng)激,提高了免疫功能,改善了肉雞與生豬的生產(chǎn)性能。兩個先天性免疫的標(biāo)記L-selectin(CD62L)和白介素8的受體(IL-8R)在嗜中性粒細胞的表達顯著提高,且在煙曲霉菌存在的情況下,差異更加顯著。血清中的各細胞因子,包括抗腫瘤因子(TNF)-α、白介素(IFN)-γ、MIP-1β及CD4+/CD8+等指標(biāo)均有改善。酵母多糖有效提高了動物疫苗抗體的滴度和高滴度的持續(xù)性。酵母多糖通過提高免疫能力和抗病能力的作用,為開發(fā)新型、環(huán)保、無公害養(yǎng)殖動物的大型傳染病防治的產(chǎn)品、減少抗生素的濫用用、降低對抗生素的耐藥性提供了新的思路。
[Abstract]:Food and medicinal fungi are important food and drug resources of human beings. It is of profound significance to study the mechanism, function and Prospect of active substances of edible and medicinal fungi, to develop and popularize the edible and medicinal fungi in depth and thus benefit mankind. Yeast, as one of the important fungi, has been applied for thousands of years. The yeast cell wall is rich in polysaccharide, low in cost and abundant in supply. It is a widely used fungal polysaccharide. The main ingredient of yeast polysaccharide is glucan and mannan oligosaccharide, which has a variety of biological activity. The HPLC method for the content of chitosan and manna oligosaccharide. In this paper, we used the intestinal bleeding syndrome (a destructive disease of high yield cows) and Aspergillus fumigatus (Aspergillus fumigatus) to study the ability of yeast polysaccharide to restore immunity to immunocompromised individuals, and to find related immune markers, and establish a related detection system. A real-time quantitative method for the detection of Aspergillus fumigatus PCR to quantitatively analyze the first study of Aspergillus fumigatus DNA. in the blood and tissue samples of dairy cows in the intestinal bleeding syndrome shows that there is a positive correlation between the occurrence of intestinal bleeding syndrome and Aspergillus fumigatus. As Aspergillus fumigatus has the characteristics of individuals with attack of immune injury (immunocomprisied), the cause of Aspergillus fumigatus is due to the characteristics of the individual. This part of this study was conducted in a sheep experiment to verify the hypothesis, and to detect the ability of fungal polysaccharides to regulate innate and acquired immune functions. The study confirmed that a sheep with dexamethasone injected during the growth period could establish an immune injury model, and daily dexamethasone was injected to reduce L- selectin and IL-1 beta. The concentration of yeast polysaccharide can restore the gene expression level of L- selectin and IL-1 beta in the immunosuppressed sheep. The yeast polysaccharide enhances the function of neutrophils by enhancing the expression of L- selectin and IL-1 beta, and inhibits the apoptosis signal and strengthens the function of neutrophils to reach the target site accurately, making it effective. The immunomodulatory function of yeast fungus polysaccharide and its anti-tumor mechanism were studied in this paper. In the experiment, we studied the effect of yeast polysaccharide on the immune function of normal mice and immunosuppressive mice, and further used the H22 tumor bearing mouse model to study the resistance of yeast polysaccharide to the immune system. The mechanism of cancer effect was analyzed by statistical analysis of tumor weight, immune organ index and related cytokine content, and the possible anti-tumor mechanism was discussed. The experimental results showed that the yeast fungus polysaccharide had anti tumor effect and increased the weight of mice and increased the immune organ index. Cyclophosphamide (CTX) was a kind of alkane. The antitumor agents of chemicals have a certain inhibitory effect on the immunity of the body, although it can effectively inhibit the tumor, but it also has a strong side effect, which causes the immune organs of the CTX group to present different degrees of atrophy and the related immune indexes are reduced, but the yeast polysaccharide has obvious regulating effect on the side effects produced by it. The mice feeding experiments showed that Yeast Polysaccharides significantly promoted the spleen and thymus organs of mice. Fungal polysaccharides could significantly improve a series of immune markers, including TNF- alpha, IFN- gamma, CRP and IL-6. These genes were expressed in intestinal cells (P0.05). ELISA was used to detect the concentration of TNF- a, IFN-, CRP and IL-6 in rat serum by ELISA. The results showed that the concentration of these cytokines increased significantly in the experimental group (P0.05). It indicated that the yeast polysaccharide promoted the immune regulation function by promoting the gene expression of cytokines. This study purify neutrophils in blood samples, and then studied the effects of Polysaccharide on the expression of TNF- a, IFN- gamma, CRP and IL-6. Yeast Polysaccharides significantly increase the expression of these cytokines (P0.05), and increase the expression of L-selectin and IL-8R in the intestinal and neutrophils (P0.05) and enhance the phagocytosis of neutrophils. The specificity of the selected markers indicates that the yeast is innate and acquired immunity, including humoral and cellular immunity. In order to explore the antitumor effect of yeast polysaccharide on the activity of tumor cells in vitro, the inhibitory rate of breast cancer MCF-7 and ovarian cancer SKOV3 cells was studied. Sugar has a certain inhibitory effect on the proliferation of two cancer cells. This study conducted experiments on Broilers and pig animals to verify the effect of Yeast Polysaccharides on the immune function in cultured animals. The infection of Aspergillus fumigatus led to high mortality and slow growth of broilers. The infection of Aspergillus fumigatus also resulted in sow diarrhoea. In the two experiments, the yeast polysaccharide (glucan and manna oligosaccharide) alleviates the stress of Aspergillus fumigatus and improves the immune function in the two groups of experiments. The production performance of Broilers and pigs was improved. The expression of two innate immune markers L-selectin (CD62L) and interleukin 8 receptor (IL-8R) was significantly increased in neutrophils, and the difference was more significant in the presence of Aspergillus fumigatus. All cell factors in the serum, including anti tumor factor (TNF) - alpha, IFN - gamma, MIP-1 beta The Yeast Polysaccharides effectively improve the titer and high titer of the animal vaccine. By improving the immune and disease resistance, yeast polysaccharides can develop new, environmentally friendly, harmless breeding animals to prevent and control large infectious diseases, reduce the abuse of antibiotics and reduce antibiotics. Drug resistance provides a new way of thinking.

【學(xué)位授予單位】：吉林農(nóng)業(yè)大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2015
【分類號】：S859.7

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