本文關(guān)鍵詞：酪蛋白酶解肽譜對樹突狀細(xì)胞成熟的影響　出處：《天津商業(yè)大學(xué)》2010年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 牛乳酪蛋白 水解多肽 先天性免疫 腸粘膜免疫系統(tǒng) 樹突狀細(xì)胞 獲得性免疫

【摘要】： 酪蛋白經(jīng)水解制得的多肽是重要的生物活性肽,具有免疫調(diào)節(jié)能力。本研究分別利用胰蛋白酶及胃蛋白酶的酶解酪蛋白,經(jīng)乙醇、超濾以及Sephadex G-25兩次分離純化后,制備兩種不同的酪蛋白水解多肽。一方面以Balb/c小鼠為實驗動物,分別將同一濃度的胰蛋白酶水解的酪蛋白多肽通過口服,腹腔注射和靜脈注射給藥。利用懸浮液體芯片系統(tǒng),檢測小鼠外周血血清中24種細(xì)胞因子的濃度變化,研究胰蛋白酶酪蛋白水解多肽對小鼠細(xì)胞因子網(wǎng)絡(luò)的影響并推測產(chǎn)生的先天免疫應(yīng)答。結(jié)果灌胃組中,大部分細(xì)胞因子的水平是升高的,抗炎細(xì)胞因子IL-5,IL-9,IL-10,MIP-1α是降低的,說明酪蛋白水解多肽促進(jìn)發(fā)炎細(xì)胞因子的合成,具有刺激免疫升高的作用。腹腔注射組與此類似。靜脈注射的酪蛋白水解多肽對細(xì)胞因子水平的影響與灌胃組、腹腔注射組相比明顯不同。因為多肽不經(jīng)過腸粘膜免疫系統(tǒng)直接進(jìn)入血液,推測通過激活多種免疫細(xì)胞內(nèi)部不同的信號途徑影響了細(xì)胞因子的基因表達(dá)。另一方面誘導(dǎo)培養(yǎng)C57BL/6雄性小鼠骨髓源的樹突狀細(xì)胞(Dendritic cells),用不同濃度的水解多肽刺激DCs,在48h后進(jìn)行流式細(xì)胞儀檢測,鑒定DCs的成熟程度,并將成熟后的DCs與T細(xì)胞共孵育96h,CCK-8檢測T細(xì)胞增殖。結(jié)果表明:兩種酶水解酪蛋白多肽均能夠刺激DCs的成熟,但刺激程度低于陽性對照LPS。而且成熟的DCs能夠刺激T細(xì)胞增殖,說明DCs具有對這些多肽的抗原遞呈能力。其上清液中的細(xì)胞因子IL-1α、IL-β、IL-2、IL-6、IL-12的分泌提高, TNF-α、IFN-γ濃度降低,促使幼稚T細(xì)胞向Th2極化。說明酪蛋白水解多肽對獲得性免疫具有信號傳遞作用。
[Abstract]:The peptides obtained by hydrolysis of casein are important bioactive peptides with immunomodulatory ability. In this study, casein was hydrolyzed by trypsin and pepsin respectively, and ethanol was used to hydrolyze casein. Two different casein hydrolysates were prepared after ultrafiltration and Sephadex G-25 purification. On the one hand, Balb/c mice were used as experimental animals. Casein peptides hydrolyzed with the same concentration of trypsin were administered orally, intraperitoneally and intravenously. To study the effect of trypsin casein hydrolysate polypeptide on cytokine network and to speculate the innate immune response in mice peripheral blood serum 24 cytokines were detected. Results in the gastric perfusion group the effect of trypsin casein hydrolysate polypeptide on the cytokine network of mice was studied. The levels of most cytokines were increased, and the anti-inflammatory cytokines IL-5, IL-9, IL-10 and MIP-1 偽 were decreased, indicating that casein hydrolyzed polypeptides promoted the synthesis of inflammatory cytokines. It has the effect of stimulating immune elevation. The intraperitoneal injection group is similar. The effect of intravenously injected casein hydrolysate polypeptide on cytokine level is similar to that in the gavage group. The intraperitoneal injection group was significantly different because peptides entered the blood without going through the intestinal mucosal immune system. It is speculated that the gene expression of cytokines is affected by activating different signaling pathways within immune cells. On the other hand, dendritic cells derived from bone marrow of C57BL / 6 male mice are induced. Dendritic cells. DCs were stimulated with different concentrations of hydrolyzed polypeptides. Flow cytometry was performed after 48 hours to identify the maturity of DCs. The mature DCs was co-incubated with T cells for 96 h. CCK-8 was used to detect the proliferation of T cells. The results showed that casein peptides hydrolyzed by two enzymes could stimulate the maturation of DCs. Moreover, mature DCs can stimulate T cell proliferation, indicating that DCs has the ability to present antigen to these polypeptides. The cytokine IL-1 偽 in the supernatant of DCs. The secretion of IL-12 was increased, and the concentration of TNF- 偽 and IFN- 緯 was decreased. The results showed that casein hydrolysate polypeptide had signal transduction effect on acquired immunity.
【學(xué)位授予單位】：天津商業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2010
【分類號】：R392

【參考文獻(xiàn)】

相關(guān)期刊論文 前8條

1 梁振,張維冰,張玉奎;CZE-ESI-MS聯(lián)用測定小肽混合物的研究[J];高等學(xué)校化學(xué)學(xué)報;2004年03期

2 張臨友,張學(xué)峰;樹突狀細(xì)胞的起源和分化[J];國外醫(yī)學(xué)(免疫學(xué)分冊);2003年01期

3 鄭華,傅偉龍;酪蛋白水解物對小鼠生長及免疫功能的影響[J];華南農(nóng)業(yè)大學(xué)學(xué)報;2000年03期

4 丁傳林;樹突狀細(xì)胞的發(fā)育、亞群及其對T細(xì)胞應(yīng)答類型的調(diào)控[J];上海免疫學(xué)雜志;2002年05期

5 崔波,程云輝,鐘秋平;毛細(xì)管電泳在多肽分離和檢測中的應(yīng)用[J];食品與機(jī)械;2004年03期

6 龐廣昌;陳慶森;胡志和;;食品是如何通過細(xì)胞因子網(wǎng)絡(luò)控制人類健康的(I)[J];食品科學(xué);2006年05期

7 龐廣昌;陳慶森;胡志和;;食品是如何通過細(xì)胞因子網(wǎng)絡(luò)控制人類健康的(II)[J];食品科學(xué);2006年06期

8 張霞,程富勝,董鵬程,程世紅,金莉;樹突狀細(xì)胞在免疫學(xué)上的研究進(jìn)展[J];中國動物保健;2003年07期

，

本文編號：1371638