本文關(guān)鍵詞：苦蕎蛋白對(duì)小鼠血脂代謝紊亂與腸道菌群調(diào)節(jié)作用的研究　出處：《上海應(yīng)用技術(shù)大學(xué)》2016年碩士論文　論文類(lèi)型：學(xué)位論文

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【摘要】：隨著人民生活水平的提高,膳食結(jié)構(gòu)發(fā)生了巨大的變化,膳食中油脂的比例不斷增多,可誘發(fā)血脂代謝紊亂與氧化應(yīng)激,影響腸道菌群平衡,危害人體健康�？嗍w富含多種功能因子,如：苦蕎蛋白,黃酮類(lèi)化合物,抗性淀粉等,其中苦蕎蛋白是一種難消化的蛋白,具有吸附膽酸鹽活性調(diào)節(jié)脂代謝的作用,但目前鮮有研究涉及苦蕎蛋白對(duì)腸道菌群影響及其與調(diào)節(jié)血脂代謝的關(guān)聯(lián)。因此,本文通過(guò)膳食誘發(fā)血脂代謝紊亂小鼠模型探究苦蕎蛋白對(duì)干預(yù)高脂代謝紊亂、氧化應(yīng)激與腸道菌群失衡作用之間的關(guān)聯(lián)。C57BL/6雄性小鼠隨機(jī)分為5組(每組9只)：空白組(Blank,AIN-93G飼料)；高脂組(HFD,10%脂肪,1%膽固醇,0.5%膽酸鈉)；苦蕎粉干預(yù)組(PBF,高蛋白含量苦蕎粉)；苦蕎蛋白干預(yù)組(BWP)；黃酮干預(yù)組(Fla,0.5%苦蕎黃酮提取物)。飼喂6w后,分析血漿與組織勻漿液的血脂血糖代謝相關(guān)生化指標(biāo)、抗氧化活性指標(biāo),并通過(guò)LC-MS、GC-MS和分子生物學(xué)技術(shù)分析腸道主要菌群及其代謝產(chǎn)物的變化。研究結(jié)果如下：(1)高脂膳食可引發(fā)小鼠血脂代謝紊亂、胰島素敏感性(ISI)顯著下降(P0.05)、氧化應(yīng)激,PBF和BWP能顯著降低血脂指標(biāo)(TC、TG、LDL-C)、血糖指標(biāo)(GLU、INS)、動(dòng)脈硬化指數(shù)(AI)和脂質(zhì)過(guò)氧化物指標(biāo)(MDA)的含量,提高HDL-C含量、ISI和組織的抗氧化活性(T-AOC、SOD、CAT),并基本恢復(fù)至空白組的水平。(2)PBF組蛋白質(zhì)和粗脂肪的表觀消化率較HFD組分別顯著降低了9.76%和4.06%,BWP組的分別顯著降低了6.93%和2.05%。PBF組和BWP組小鼠糞便中總膽汁酸和膽固醇的排泄較HFD組顯著增加,且腸道內(nèi)容物中乙酸、丙酸、丁酸含量顯著升高。相關(guān)性分析表明：總膽汁酸和膽固醇的排泄量與HDL-C和ISI呈顯著正相關(guān)。表明苦蕎蛋白質(zhì)難消化,未消化的蛋白質(zhì)可吸附膽酸與固醇排除體外,可能從而干預(yù)了血脂代謝紊亂。(3)通過(guò)平板計(jì)數(shù)以及PCR技術(shù)對(duì)各組小鼠的腸道主要菌群(雙歧桿菌、乳酸菌、大腸桿菌、腸球菌)進(jìn)行檢測(cè),二者結(jié)果的變化趨勢(shì)基本一致,即PBF組與BWP組的有益菌(雙歧桿菌、乳酸菌、腸球菌)數(shù)量均顯著高于HFD組,有害菌(大腸桿菌)數(shù)量則顯著低于HFD組。相關(guān)性分析表明：腸道中雙歧桿菌數(shù)量與血脂指標(biāo)(TC、TG、LDL-C)呈顯著負(fù)相關(guān),大腸桿菌數(shù)量與腸道的MDA呈顯著正相關(guān)。表明苦蕎蛋白具有調(diào)節(jié)高脂膳食小鼠腸道菌群失衡的作用,可能從而一定程度抑制血脂的紊亂及組織的氧化應(yīng)激。綜上所述,較難消化的苦蕎蛋白可吸附膽酸與膽固醇促進(jìn)其排泄,從而即調(diào)節(jié)血脂代謝又調(diào)節(jié)腸道菌群平衡,而腸道菌群中益生菌比例增多有害菌比例的降低又對(duì)血脂代謝調(diào)節(jié)與抑制氧化應(yīng)激有著一定促進(jìn)作用。因此,苦蕎蛋白對(duì)血脂代謝調(diào)節(jié)作用可能與其促進(jìn)膽酸排泄、調(diào)節(jié)腸道菌群平衡、改善氧化應(yīng)激等方面作用途徑有著密切關(guān)系。
[Abstract]:With the improvement of people's living standard, the dietary structure has changed greatly, and the proportion of fat in diet is increasing, which can induce lipid metabolism disorder and oxidative stress, and affect the balance of intestinal flora. Tartary buckwheat is rich in many functional factors, such as Tartary buckwheat protein, flavonoids, resistant starch and so on, among which Tartary buckwheat protein is an indigestible protein. It has the function of adsorbing cholate activity to regulate lipid metabolism, but there are few studies on the effect of Tartary buckwheat protein on intestinal flora and its relationship with the regulation of lipid metabolism. In this paper, the effect of Tartary buckwheat protein (Tartary buckwheat protein) on hyperlipidemia was investigated in mice model of hyperlipidemia induced by diet. The relationship between oxidative stress and intestinal flora imbalance. C57BL / 6 male mice were randomly divided into 5 groups (9 rats in each group): blank group (Blanck AIN-93G feed); In the hyperlipidemia group, 10% HFDF 10% fat and 1% cholesterol 0. 5% sodium cholate; Tartary buckwheat powder intervention group, high protein content of Tartary buckwheat powder; Tartary buckwheat protein intervention group (BWPN); The flavonoid intervention group was fed with 0.5% Tartary buckwheat flavone extract for 6 weeks. The biochemical indexes of blood lipids metabolism and antioxidant activity of plasma and tissue homogenate were analyzed by LC-MS. GC-MS and molecular biology techniques were used to analyze the changes of intestinal microflora and its metabolites. The results were as follows: 1) High fat diet could induce lipid metabolism disorder in mice. Insulin sensitivity index (ISI) decreased significantly (P 0.05), and oxidative stress and BWP significantly decreased serum lipids, TGG, LDL-C and GLU. The contents of HDL-C and T-AOC SOD in tissues were increased by the contents of HDL-C and lipid peroxides (MDA), the arteriosclerotic index (AII) and the antioxidant activity (T-AOC) of the tissue. The apparent digestibility of protein and crude fat in HFD group was significantly lower than that in HFD group by 9.76% and 4.06% respectively. The excretion of total bile acid and cholesterol in BWP group was significantly lower than that in HFD group. The excretion of total bile acid and cholesterol in the feces of PBF group and BWP group were significantly higher than that in HFD group, and acetic acid in intestinal contents. The content of propionic acid and butyric acid was significantly increased. Correlation analysis showed that the excretion of total bile acid and cholesterol was positively correlated with HDL-C and ISI, indicating that Tartary buckwheat protein was indigestible. The undigested protein can adsorb cholic acid and steroid to exclude in vitro, which may interfere with dyslipidemia. The main intestinal flora (Bifidobacterium) of mice in each group was determined by plate counting and PCR technique. Lactic acid bacteria, Escherichia coli, enterococcus were detected, the results of the two changes are basically the same trend, that is, PBF group and BWP group of beneficial bacteria (Bifidobacterium, lactic acid bacteria). The number of Enterococcus was significantly higher than that of HFD group, and the number of harmful bacteria (Escherichia coli) was significantly lower than that of HFD group. LDL-C was negatively correlated, and the number of Escherichia coli was positively correlated with intestinal MDA, indicating that Tartary buckwheat protein could regulate intestinal microflora imbalance in high-fat diet mice. In conclusion, the more difficult to digest Tartary buckwheat protein can adsorb cholic acid and cholesterol to promote its excretion. Thus regulating the metabolism of blood lipids and regulating the balance of intestinal flora, and the increase of the proportion of probiotics in the intestinal bacteria proportion of harmful bacteria in the proportion of lipid metabolism regulation and inhibition of oxidative stress has a certain role in promoting. The effects of Tartary buckwheat protein on lipid metabolism may be closely related to the regulation of cholic acid excretion, the regulation of intestinal microflora balance and the improvement of oxidative stress.
【學(xué)位授予單位】：上海應(yīng)用技術(shù)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類(lèi)號(hào)】：R151
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本文編號(hào)：1368487