本文選題：卵黃高磷蛋白磷酸肽 + 生長性能��； 參考：《四川農(nóng)業(yè)大學(xué)》2009年碩士論文

【摘要】： 為了初步探索卵黃高磷蛋白磷酸肽(PPP)是否具有免疫作用,本研究在制備了PPP的基礎(chǔ)上,考察了PPP對小鼠體內(nèi)外免疫功能的影響,并初步分析PPP影響機體免疫功能的機理。本研究包括以下三個試驗。 試驗一,PPP的制備。本試驗首先從雞蛋黃中提取卵黃高磷蛋白(PV),經(jīng)胰蛋白酶水解,水解液經(jīng)截留分子量3 kDa和1 kDa的超濾膜超濾,取分子量在1 kDa-3 kDa之間的截留液冷凍干燥得到PPP。經(jīng)SDS-PAGE電泳測PV的純度,用N/P(摩爾比)檢測PPP的磷酸絲氨酸殘基密度。結(jié)果表明:所制備的PV樣品與純品電泳模式相近,在1 kDa-3 kDa范圍內(nèi)存在PPP電泳條帶,且N/P比例為5.02。說明所提取PV樣品相對較純,PPP樣品含有磷酸絲氨酸殘基。 試驗二,PPP對小鼠體外免疫功能的影響。為了評價PPP對細胞增殖、吞噬功能、細胞因子以及免疫球蛋白生成的影響,分離小鼠的脾淋巴細胞和腹腔巨噬細胞并進行培養(yǎng)。結(jié)果顯示:添加LPS或ConA后,顯著地促進了細胞的增殖、細胞吞噬功能、細胞因子的分泌以及免疫球蛋白的生成(p＜0.05)。終濃度為0.02 mg/mL的PV組不能顯著地影響細胞功能(p＞0.05)。然而,終濃度為0.02 mg/mL的PPP組和0.1 mg/mL PPP組顯著地提高了刺激指數(shù)(SI)和吞噬指數(shù)(PI),以及IgA、IL-1β、IL-2和IL-6的生成。LPS或ConA誘導(dǎo)組比無絲裂原組刺激作用顯著增強。結(jié)果表明,PPP顯著提高了SI、PI、細胞因子以及免疫球蛋白生成的影響,無絲裂原組相比,LPS或ConA誘導(dǎo)下,以上各指標(biāo)均顯著提高。 試驗三,PPP對小鼠體內(nèi)免疫功能的影響。為了確定胃飼PPP對小鼠免疫功能的影響,將48只小鼠隨機分到3個處理中,每個處理設(shè)4個重復(fù),每個處理的小鼠在基礎(chǔ)日糧的基礎(chǔ)上分別隔天灌胃0.2mL生理鹽水(對照組)、PPP和PV溶液,按體重喂給0.03mg/(g·d)。14天后取樣確定小鼠的生長性能和免疫功能。結(jié)果顯示:與對照組和PV組相比,PPP組小鼠的體增重極顯著提高(p＜0.01),血清總蛋白、白蛋白、球蛋白和葡萄糖均升高,但差異不顯著(p＞0.05),而血清尿素氮、總膽固醇和甘油三酯呈下降趨勢,脾臟指數(shù)和胸腺指數(shù)呈顯著提高(p＜0.05)。PPP組的SI和PI顯著提高(p＜0.05),血清和脾淋巴細胞培養(yǎng)液中的IgA、IL-2和IL-6濃度,以及血清和腹腔巨噬細胞培養(yǎng)液中的IL-1β均顯著地上升(p＜0.05)。PV組與對照組相比,這些指標(biāo)均無顯著變化(p＞0.05)。在LPS或ConA誘導(dǎo)下,培養(yǎng)液上清中的SI、PI、IgA、IL-1β、IL-2和IL-6均有顯著的提高(p＜0.05)。結(jié)果表明,胃飼PPP不僅提高了體增重,脾臟指數(shù)和胸腺指數(shù),還顯著增強了小鼠SI和PI,血清和脾淋巴細胞培養(yǎng)液中的IgA、IL-2和IL-6濃度,以及血清和腹腔巨噬細胞培養(yǎng)液中的IL-1β濃度。與無絲裂原組相比,LPS或ConA誘導(dǎo)組以上各指標(biāo)均顯著提高。 綜合以上各試驗可看出,制備所得的PPP不僅提高了小鼠的生長性能,還增強了小鼠體內(nèi)外免疫功能。
[Abstract]:In order to explore the immune effect of phosphopeptide (PPP) on yolk high phosphorous protein (PPP), the effect of PPP on immune function of mice in vivo and in vitro was investigated on the basis of preparation of PPP, and the mechanism of PPP affecting immune function in vivo and in vitro was preliminarily analyzed. This study includes the following three experiments. The preparation of PPP. In this experiment, egg yolk high phosphorous protein (PV) was extracted from egg yolk. The protein was hydrolyzed by trypsin, and then purified by ultrafiltration membrane with molecular weight of 3 kDa and 1 kDa. PPP was obtained by freeze-drying of the retention solution with molecular weight between 1 kDa and 3 kDa. The purity of PV was measured by SDS-PAGE, and the PPP's serine residue density was measured by N / P (molar ratio). The results showed that the electrophoretic patterns of the prepared PV samples were similar to those of the pure samples. PPP bands were found in the range of 1 kDa-3 kDa, and the ratio of N / P was 5.02. The results showed that the extracted PV samples contained relatively pure PPP containing phosphate serine residues. The effect of PPP on the immune function of mice in vitro. To evaluate the effects of PPP on cell proliferation, phagocytosis, cytokine and immunoglobulin production, spleen lymphocytes and peritoneal macrophages of mice were isolated and cultured. The results showed that LPS or Cona significantly promoted cell proliferation, phagocytosis, cytokine secretion and immunoglobulin production (p < 0.05). PV with 0.02 mg / mL final concentration did not significantly affect cell function (p > 0.05). However, the stimulation index (SI) and phagocytic index (Pi) were significantly increased in the PPP group with 0.02 mg / mL final concentration and 0.1 mg / mL PPP group, as well as the production of IL-2 and IL-6 in IgA IL-1 尾, IL 2 and IL 6 production. LPS or Cona induced stimulation was significantly higher than that in the mitogen free group. The results showed that PPP significantly increased the production of SIPI, cytokines and immunoglobulin, and the above indexes were significantly higher in mitogen free group than that in LPS or Cona group. The effect of PPP on immune function in mice. In order to determine the effect of gastric feeding PPP on immune function of mice, 48 mice were randomly divided into 3 treatments, each of which was divided into 4 replicates. Each treated mice were fed with 0.2 mL normal saline solution (control group) every other day on the basis of basal diet. The growth performance and immune function of the mice were determined by 0.03mg/ (g d). 14 days later, the growth performance and immune function of the mice were determined. The results showed that compared with the control group and PV group, the body weight gain of PPP group was significantly increased (p < 0.01), the serum total protein, albumin, globulin and glucose were all increased, but the difference was not significant (p > 0.05), but the serum urea nitrogen. Total cholesterol and triglyceride decreased, spleen index and thymus index increased significantly (p < 0.05). The SI and Pi of PPP group increased significantly (p < 0.05). IL-1 尾 in serum and peritoneal macrophage culture medium were significantly increased (p < 0.05). Compared with control group, these indexes had no significant change in PV group (p > 0.05). Under the induction of LPS or Cona, the levels of IL 2 and IL 6 in the supernatant of Sipi Pia IgA 尾, IL 2 and IL 6 were significantly increased (p < 0 05). The results showed that PPP not only increased the body weight gain, spleen index and thymus index, but also significantly increased the levels of IgA, IL-2 and IL-6 in serum and splenic lymphocyte culture medium, and IL-1 尾 in serum and peritoneal macrophage culture medium. The above indexes were significantly higher in LPS or Cona-induced group than in non-mitogen group. From the above experiments, we can see that the prepared PPP not only improves the growth performance of mice, but also enhances the immune function of mice in vivo and in vitro.
【學(xué)位授予單位】：四川農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2009
【分類號】：R392.1

【引證文獻】

相關(guān)期刊論文 前1條

1 丁進鋒;蘇秀榕;李妍妍;高翔;岳福鵬;蔣鑫;;海蜇膠原蛋白肽的免疫活性的研究[J];水產(chǎn)科學(xué);2011年06期

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本文編號：2071345