腸道菌群介導(dǎo)海帶多糖干預(yù)高脂飲食誘導(dǎo)小鼠胰島素抵抗的調(diào)控機(jī)制
發(fā)布時(shí)間：2018-07-14 10:54
【摘要】：海帶(Laminaria japonica)是一種常見的海洋蔬菜,多糖是其主要的生物活性物質(zhì)。課題組前期研究發(fā)現(xiàn),海帶多糖可以改善由高脂飲食引起的小鼠胰島素抵抗(Insulin resistance)。已有研究表明,腸道菌群(gut microbiota)產(chǎn)生的LPS是胰島素抵抗病理形成的主要機(jī)制之一�；诖�,本論文以一種化學(xué)結(jié)構(gòu)和生物活性明確的海帶多糖均一組分LJP61A為實(shí)驗(yàn)材料,重點(diǎn)研究海帶多糖通過腸道菌群介導(dǎo)抑制高脂暴露誘發(fā)胰島素抵抗的調(diào)控機(jī)制。論文獲得的實(shí)驗(yàn)結(jié)果主要有:(1)LJP61A可以顯著改善小鼠由高脂飲食引起的胰島素抵抗:HFD高脂飲食會引起小鼠胰島素敏感性降低、葡萄糖耐量下降,并產(chǎn)生胰島素抵抗。胰島素耐量實(shí)驗(yàn)與口服葡萄糖耐量實(shí)驗(yàn)的結(jié)果表明LJP61A可以顯著改善小鼠的胰島素敏感性及葡萄糖耐受性。此外,LJP61A能夠顯著降低小鼠空腹葡萄糖含量、胰島素含量以及HOMA-IR值。通過western-blot分析,我們還發(fā)現(xiàn)海帶多糖LJP61A能夠顯著降低肝臟組織與脂肪組織中p-AKT、p-IRS1的蛋白表達(dá)水平。這些結(jié)果均表明LJP61A可以顯著改善高脂飲食暴露誘發(fā)的胰島素抵抗。(2)LJP61A可以顯著抑制高脂飲食誘導(dǎo)胰島素抵抗小鼠的炎癥反應(yīng):HFD會導(dǎo)致炎癥因子的過量分泌,使得機(jī)體產(chǎn)生持續(xù)的低度炎癥,而這種炎癥反應(yīng)是導(dǎo)致機(jī)體產(chǎn)生胰島素抵抗的關(guān)鍵因素之一。本實(shí)驗(yàn)采用高脂飲食HFD誘導(dǎo)C57BL小鼠胰島素抵抗的實(shí)驗(yàn)?zāi)Ｐ?研究了LJP61A對小鼠炎癥反應(yīng)的干預(yù)作用。通過RT-qPCR分析,我們發(fā)現(xiàn)LJP61A顯著改善了肝臟組織與脂肪組織中TNF-α、IL-6、IL-10、IL-1β、PAI-1mRNA表達(dá)水平,以及血清中TNF-α、IL-6、IL-1β的含量,表明LJP61A可以下調(diào)NFκB和MAPK通路下游關(guān)鍵效應(yīng)分子的表達(dá)。通過western-blot分析,我們發(fā)現(xiàn)LJP61A能夠顯著降低肝臟組織與脂肪組織中p-NFκBp65、p-P38、p-JNK、p-IκB、p-ERK1/2的蛋白表達(dá)水平,說明LJP61A抑制了調(diào)控因子NF-κB信號通路,并通過抑制JNK、ERK、P38的活化而抑制MAPK信號通路。這些結(jié)果均表明LJP61A可以顯著抑制高脂飲食誘導(dǎo)胰島素抵抗小鼠的炎癥反應(yīng)。(3)LJP61A能顯著下調(diào)高脂飲食誘導(dǎo)的小鼠腸道通透性及內(nèi)毒素血癥:HFD高脂飲食會導(dǎo)致小鼠腸道通透性的增加,進(jìn)而引起內(nèi)毒素血癥以及慢性炎癥。本次實(shí)驗(yàn)結(jié)果表明,海帶多糖LJP61A可以顯著增大腸道閉合蛋白(Occludin)、閉鎖連接蛋白(ZO-1)的表達(dá)量,以及Occludin、ZO-1 mRNA的表達(dá)水平。另外,LJP61A顯著減少了HFD小鼠血清中LPS的含量,并且降低了肝臟組織與脂肪組織中TLR4蛋白的表達(dá)量。這些結(jié)果表明LJP61A可以顯著下調(diào)高脂飲食誘導(dǎo)的小鼠腸道通透性及內(nèi)毒素血癥。(4)LJP61A對高脂飲食改變小鼠腸道菌群構(gòu)成具有干預(yù)作用:HFD飲食會導(dǎo)致腸道菌群的失調(diào)以及益生菌數(shù)量的下降,進(jìn)而引起小鼠腸道緊密性發(fā)生變化。本次試驗(yàn)發(fā)現(xiàn)高脂飲食顯著提高了Firmicutes-to-Bacteroidetes比例,且LJP61A干預(yù)濃度為200mg/kg/day時(shí)厚壁菌門細(xì)菌的含量最低,擬桿菌門含量最高。另外,LJP61A的干預(yù)使Akkermansia、Bifidobacterium、Lactobacillus等腸道益生菌的相對豐度顯著提高。這些結(jié)果表明LJP61A對高脂飲食改變小鼠腸道菌群構(gòu)成具有干預(yù)作用,可使其菌群結(jié)構(gòu)更加接近于正常對照組小鼠。以上結(jié)果表明,LJP61A通過調(diào)節(jié)HFD誘導(dǎo)小鼠的腸道菌群結(jié)構(gòu),改善小鼠腸道通透性以及內(nèi)毒素血癥,調(diào)節(jié)炎癥反應(yīng),從而達(dá)到干預(yù)胰島素抵抗的目的。
[Abstract]:Laminaria japonica is a common marine vegetable and polysaccharide is its main bioactive substance. Previous studies have found that kelp polysaccharide can improve insulin resistance (Insulin resistance) caused by high fat diet (Insulin resistance). Studies have shown that the LPS produced by the intestinal flora (gut microbiota) is the pathology of insulin resistance. In this paper, based on this, this paper uses a chemical structure and biological activity clear LJP61A as the experimental material, focusing on the regulation mechanism of inhibiting high fat exposure through the intestinal flora mediated by the intestinal flora. The results of the paper are as follows: (1) LJP61A can be significant Improve insulin resistance induced by high fat diet in mice: HFD high fat diet can cause insulin sensitivity to decrease, glucose tolerance and insulin resistance. The results of insulin tolerance test and oral glucose tolerance test show that LJP61A can significantly improve insulin sensitivity and glucose tolerance in rats. LJP61A can significantly reduce the fasting glucose content, insulin content and HOMA-IR value in mice. Through Western-blot analysis, we also found that the Laminaria Polysaccharide LJP61A significantly reduced the protein expression levels of p-AKT and p-IRS1 in the liver and adipose tissues. These results suggest that LJP61A can significantly improve the exposure to high fat diet exposure. Insulin resistance. (2) LJP61A can significantly inhibit the inflammatory response induced by high fat diet induced insulin resistance in mice: HFD leads to excessive secretion of inflammatory factors and causes the body to produce persistent low degree of inflammation, which is one of the key factors leading to the body's insulin resistance. This experiment uses a high fat diet HFD to induce C57BL The experimental model of insulin resistance in mice was used to study the effect of LJP61A on the inflammatory response in mice. Through RT-qPCR analysis, we found that LJP61A significantly improved the level of TNF- alpha, IL-6, IL-10, IL-1 beta, PAI-1mRNA expression in the liver tissue and adipose tissue, as well as the content of TNF- alpha, IL-6, IL-1 beta in the serum. Through Western-blot analysis, we found that LJP61A can significantly reduce the protein expression level of p-NF kappa Bp65, p-P38, p-JNK, p-I kappa B, and p-ERK1/2 in the liver and adipose tissue. It shows that LJP61A inhibits the NF- kappa B signal pathway, and inhibits the activation by inhibiting the activation. These results all indicate that LJP61A can significantly inhibit the inflammatory response in insulin resistant mice induced by high fat diet. (3) LJP61A can significantly reduce the intestinal permeability and endotoxemia in mice induced by high fat diet. HFD high fat diet may lead to the increase of intestinal permeability in mice, and then cause endotoxemia and chronic inflammation. The results showed that LJP61A could significantly increase the intestinal closed protein (Occludin), the expression of interlocking connexin (ZO-1), and the expression level of Occludin and ZO-1 mRNA. In addition, LJP61A significantly reduced the content of LPS in the serum of HFD mice, and reduced the expression of TLR4 protein in the liver and adipose tissues. The results showed that LJP61A could significantly reduce the intestinal permeability and endotoxemia in mice induced by high fat diet. (4) LJP61A has an intervention effect on the formation of intestinal flora in mice with high fat diet. The HFD diet will lead to the imbalance of intestinal flora and the decrease of the number of probiotics, and then lead to the change of intestinal tightness in mice. In the present high fat diet, the proportion of Firmicutes-to-Bacteroidetes was significantly increased, and the content of bacilli was the lowest when the concentration of LJP61A was 200mg/kg/day, and the highest content of the bacilli was found. In addition, the intervention of LJP61A significantly increased the relative abundance of intestinal probiotics, such as Akkermansia, Bifidobacterium, Lactobacillus and so on. These results showed LJP61A. The results showed that LJP61A could improve the intestinal permeability and endotoxemia and regulate the inflammatory response by regulating the intestinal microflora structure of mice by regulating HFD, and thus interfering with insulin resistance. The purpose.
【學(xué)位授予單位】：合肥工業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R151
本文編號：2121417
[Abstract]:Laminaria japonica is a common marine vegetable and polysaccharide is its main bioactive substance. Previous studies have found that kelp polysaccharide can improve insulin resistance (Insulin resistance) caused by high fat diet (Insulin resistance). Studies have shown that the LPS produced by the intestinal flora (gut microbiota) is the pathology of insulin resistance. In this paper, based on this, this paper uses a chemical structure and biological activity clear LJP61A as the experimental material, focusing on the regulation mechanism of inhibiting high fat exposure through the intestinal flora mediated by the intestinal flora. The results of the paper are as follows: (1) LJP61A can be significant Improve insulin resistance induced by high fat diet in mice: HFD high fat diet can cause insulin sensitivity to decrease, glucose tolerance and insulin resistance. The results of insulin tolerance test and oral glucose tolerance test show that LJP61A can significantly improve insulin sensitivity and glucose tolerance in rats. LJP61A can significantly reduce the fasting glucose content, insulin content and HOMA-IR value in mice. Through Western-blot analysis, we also found that the Laminaria Polysaccharide LJP61A significantly reduced the protein expression levels of p-AKT and p-IRS1 in the liver and adipose tissues. These results suggest that LJP61A can significantly improve the exposure to high fat diet exposure. Insulin resistance. (2) LJP61A can significantly inhibit the inflammatory response induced by high fat diet induced insulin resistance in mice: HFD leads to excessive secretion of inflammatory factors and causes the body to produce persistent low degree of inflammation, which is one of the key factors leading to the body's insulin resistance. This experiment uses a high fat diet HFD to induce C57BL The experimental model of insulin resistance in mice was used to study the effect of LJP61A on the inflammatory response in mice. Through RT-qPCR analysis, we found that LJP61A significantly improved the level of TNF- alpha, IL-6, IL-10, IL-1 beta, PAI-1mRNA expression in the liver tissue and adipose tissue, as well as the content of TNF- alpha, IL-6, IL-1 beta in the serum. Through Western-blot analysis, we found that LJP61A can significantly reduce the protein expression level of p-NF kappa Bp65, p-P38, p-JNK, p-I kappa B, and p-ERK1/2 in the liver and adipose tissue. It shows that LJP61A inhibits the NF- kappa B signal pathway, and inhibits the activation by inhibiting the activation. These results all indicate that LJP61A can significantly inhibit the inflammatory response in insulin resistant mice induced by high fat diet. (3) LJP61A can significantly reduce the intestinal permeability and endotoxemia in mice induced by high fat diet. HFD high fat diet may lead to the increase of intestinal permeability in mice, and then cause endotoxemia and chronic inflammation. The results showed that LJP61A could significantly increase the intestinal closed protein (Occludin), the expression of interlocking connexin (ZO-1), and the expression level of Occludin and ZO-1 mRNA. In addition, LJP61A significantly reduced the content of LPS in the serum of HFD mice, and reduced the expression of TLR4 protein in the liver and adipose tissues. The results showed that LJP61A could significantly reduce the intestinal permeability and endotoxemia in mice induced by high fat diet. (4) LJP61A has an intervention effect on the formation of intestinal flora in mice with high fat diet. The HFD diet will lead to the imbalance of intestinal flora and the decrease of the number of probiotics, and then lead to the change of intestinal tightness in mice. In the present high fat diet, the proportion of Firmicutes-to-Bacteroidetes was significantly increased, and the content of bacilli was the lowest when the concentration of LJP61A was 200mg/kg/day, and the highest content of the bacilli was found. In addition, the intervention of LJP61A significantly increased the relative abundance of intestinal probiotics, such as Akkermansia, Bifidobacterium, Lactobacillus and so on. These results showed LJP61A. The results showed that LJP61A could improve the intestinal permeability and endotoxemia and regulate the inflammatory response by regulating the intestinal microflora structure of mice by regulating HFD, and thus interfering with insulin resistance. The purpose.
【學(xué)位授予單位】：合肥工業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R151
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