小鼠腸道菌群構(gòu)成與肥胖易發(fā)性的研究
發(fā)布時(shí)間：2018-07-23 17:47
【摘要】：人體腸道中龐大的微生物群落與宿主互惠共生,在機(jī)體能量的獲取、脂肪代謝的調(diào)控以及慢性炎癥的發(fā)生等方面起重要作用,與肥胖和代謝綜合征的發(fā)生發(fā)展密切相關(guān)。小鼠攝取高脂膳食可產(chǎn)生肥胖抵抗(DIO-R)和肥胖易感(DIO-P)兩種表型,探究其腸道菌群有利于闡明肥胖產(chǎn)生機(jī)制,對(duì)提出干預(yù)肥胖的營(yíng)養(yǎng)手段具有重要意義。將C57BL/6小鼠隨機(jī)分為對(duì)照組(標(biāo)準(zhǔn)飼料飼養(yǎng),NC)和高脂飼料組,7周后將高脂組分為肥胖抵抗組(DIO-R)和肥胖易感組(DIO-P)。在實(shí)驗(yàn)起始(0周)、肥胖發(fā)展的早期(7周)、中期(17周)和末期(27周),利用末端限制性片段長(zhǎng)度多態(tài)性(T-RFLP)和定量PCR檢測(cè)糞便菌群結(jié)構(gòu),并測(cè)定短鏈脂肪酸含量;在7、17和27周分別處死部分小鼠,分析血液脂多糖(LPS)和炎性因子水平,檢測(cè)回腸上皮TLR4、GPR41、Fiaf、Zo-1、Occludin和Muc2基因的表達(dá)情況。結(jié)果顯示:DIO-P組7周后體重顯著高于另兩組(P0.05),DIO-R組體重在中后期逐漸高于NC組(P0.05);DIO-P組腸道菌群與NC組和DIO-R組差異明顯,多樣性和穩(wěn)定性下降,同時(shí)乳桿菌(Lactobacillus)、雙歧桿菌(Bifidobacterium)、Akkermansia muciniphila豐度下降,而脫硫弧菌(Desulfovibrio)、金黃色葡萄球菌(Staphylococcus aureus)、陰溝腸桿菌(Enterobacter cloacae)豐度上升;DIO-R組這幾種菌的含量在早期變化不明顯,中后期逐漸趨向DIO-P組特征。試驗(yàn)起始時(shí)DIO-R組和DIO-P組小鼠腸道菌群構(gòu)成存在顯著差異,與肥胖發(fā)生具有顯著相關(guān)性。與NC組相比,DIO-P組糞便乙酸、丁酸含量在7周后顯著降低,而丙酸顯著升高(P0.05),在前期與DIO-R組差異顯著(P0.05),而后期無顯著差異。DIO-P組血液LPS在7周后顯著高于NC組(P0.05),在7周和17周時(shí)顯著高于DIO-R組(P0.05)�；啬cTLR4(LPS受體)和GPR41(短鏈脂肪酸的受體)基因表達(dá)在DIO-P組顯著高于DIO-R組(P0.05),說明腸道可能出現(xiàn)炎性反應(yīng)和短鏈脂肪酸代謝異常;Fiaf介導(dǎo)信號(hào)參與脂肪分解、抑制脂肪合成,DIO-P組該基因表達(dá)顯著低于DIO-R組(P0.05)。Zo-1、Occludin和Muc2三個(gè)基因與腸粘膜屏障完整性有關(guān),DIO-P組這三個(gè)基因的表達(dá)顯著低于DIO-R組(P0.05),說明肥胖易感組的腸道屏障可能受損。ELISA分析顯示DIO-P組小鼠血液促炎性細(xì)胞因子顯著上升(P0.05),而抗炎性細(xì)胞因子含量顯著下降(P0.05),說明發(fā)生了慢性系統(tǒng)性炎癥�？傊�,本研究說明小鼠在高脂膳食下肥胖的易發(fā)性與腸道菌群有密切的聯(lián)系,腸道菌群影響肥胖易感的機(jī)制可能與損傷腸黏膜屏障、誘發(fā)炎性反應(yīng)和擾亂脂代謝有關(guān)。
[Abstract]:The large microbial community in the human intestine is symbiotic with the host, which plays an important role in the acquisition of body energy, the regulation of fat metabolism and the occurrence of chronic inflammation, which is closely related to the occurrence and development of obesity and metabolic syndrome. Two phenotypes of obesity resistance (DIO-R) and obesity susceptibility (DIO-P) can be produced in mice fed with high fat diet. It is important to explore the intestinal flora of mice to elucidate the mechanism of obesity and to propose nutritional intervention for obesity. C57BL/6 mice were randomly divided into control group (standard diet feeding NC) and high-fat diet group after 7 weeks. The high-fat group was divided into obese resistance group (DIO-R) and obesity susceptible group (DIO-P). At the beginning of the experiment (0 weeks), the early stage of obesity (7 weeks), the middle stage (17 weeks) and the end stage (27 weeks), the fecal microflora structure was detected by terminal restriction fragment length polymorphism (T-RFLP) and quantitative PCR, and the content of short chain fatty acids was measured. The blood lipopolysaccharide (LPS) and inflammatory cytokines were analyzed, and the expression of TLR4GPR41 Fiafo Zo-1Occludin and Muc2 genes in ileum epithelium were detected at 7 ~ (th) ~ (th) week and 27 ~ (th) week, respectively. The results showed that the body weight of the DIO-P group was significantly higher than that of the other two groups after 7 weeks (P0.05). The intestinal flora of the DIO-P group was significantly higher than that of the NC group (P0.05), the diversity and stability of the intestinal flora of the DIO-P group were significantly lower than those of the NC group and the DIO-R group, and the abundance of (Bifidobacterium) Akkermansia muciniphila in the DIO-R group was decreased. However, the (Enterobacter cloacae) abundance of (Desulfovibrio), Staphylococcus aureus (Staphylococcus aureus), increased in DIO-R group, and the content of these bacteria did not change obviously at the early stage, but gradually tended to the characteristics of DIO-P group in the middle and late stage. At the beginning of the experiment, there was a significant difference between DIO-R group and DIO-P group in the composition of intestinal flora, and there was a significant correlation between the composition of intestinal flora and obesity. Compared with NC group, the content of acetic acid and butyric acid in feces of DIO-P group decreased significantly after 7 weeks. But propionic acid significantly increased (P0.05), in the Prophase and DIO-R group significant difference (P0.05), but no significant difference in late stage. DIO-P group was significantly higher than NC group after 7 weeks (P0.05), at 7 weeks and 17 weeks significantly higher than DIO-R group (P0.05). The expression of TLR4 (LPS receptor) and GPR41 (short chain fatty acid receptor) gene in ileum was significantly higher in DIO-P group than in DIO-R group (P0.05). The expression of this gene in the DIO-P group was significantly lower than that in the DIO-R group (P0.05). Zo-1Occludin and Muc2 were associated with the integrity of intestinal mucosal barrier. The expression of these three genes in the DIO-P group was significantly lower than that in the DIO-R group (P0.05), indicating that the intestinal barrier might be damaged in the obese susceptible group. Elisa analysis showed that the blood pro-inflammatory cytokines in DIO-P group increased significantly (P0.05), while the content of anti-inflammatory cytokines decreased significantly (P0.05), indicating the occurrence of chronic systemic inflammation. In conclusion, this study shows that the susceptibility to obesity in mice under high fat diet is closely related to intestinal flora. The mechanism of intestinal flora affecting obesity susceptibility may be related to injury of intestinal mucosal barrier, inflammatory response and disturbance of lipid metabolism.
【學(xué)位授予單位】：江南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R589.2
本文編號(hào)：2140163
[Abstract]:The large microbial community in the human intestine is symbiotic with the host, which plays an important role in the acquisition of body energy, the regulation of fat metabolism and the occurrence of chronic inflammation, which is closely related to the occurrence and development of obesity and metabolic syndrome. Two phenotypes of obesity resistance (DIO-R) and obesity susceptibility (DIO-P) can be produced in mice fed with high fat diet. It is important to explore the intestinal flora of mice to elucidate the mechanism of obesity and to propose nutritional intervention for obesity. C57BL/6 mice were randomly divided into control group (standard diet feeding NC) and high-fat diet group after 7 weeks. The high-fat group was divided into obese resistance group (DIO-R) and obesity susceptible group (DIO-P). At the beginning of the experiment (0 weeks), the early stage of obesity (7 weeks), the middle stage (17 weeks) and the end stage (27 weeks), the fecal microflora structure was detected by terminal restriction fragment length polymorphism (T-RFLP) and quantitative PCR, and the content of short chain fatty acids was measured. The blood lipopolysaccharide (LPS) and inflammatory cytokines were analyzed, and the expression of TLR4GPR41 Fiafo Zo-1Occludin and Muc2 genes in ileum epithelium were detected at 7 ~ (th) ~ (th) week and 27 ~ (th) week, respectively. The results showed that the body weight of the DIO-P group was significantly higher than that of the other two groups after 7 weeks (P0.05). The intestinal flora of the DIO-P group was significantly higher than that of the NC group (P0.05), the diversity and stability of the intestinal flora of the DIO-P group were significantly lower than those of the NC group and the DIO-R group, and the abundance of (Bifidobacterium) Akkermansia muciniphila in the DIO-R group was decreased. However, the (Enterobacter cloacae) abundance of (Desulfovibrio), Staphylococcus aureus (Staphylococcus aureus), increased in DIO-R group, and the content of these bacteria did not change obviously at the early stage, but gradually tended to the characteristics of DIO-P group in the middle and late stage. At the beginning of the experiment, there was a significant difference between DIO-R group and DIO-P group in the composition of intestinal flora, and there was a significant correlation between the composition of intestinal flora and obesity. Compared with NC group, the content of acetic acid and butyric acid in feces of DIO-P group decreased significantly after 7 weeks. But propionic acid significantly increased (P0.05), in the Prophase and DIO-R group significant difference (P0.05), but no significant difference in late stage. DIO-P group was significantly higher than NC group after 7 weeks (P0.05), at 7 weeks and 17 weeks significantly higher than DIO-R group (P0.05). The expression of TLR4 (LPS receptor) and GPR41 (short chain fatty acid receptor) gene in ileum was significantly higher in DIO-P group than in DIO-R group (P0.05). The expression of this gene in the DIO-P group was significantly lower than that in the DIO-R group (P0.05). Zo-1Occludin and Muc2 were associated with the integrity of intestinal mucosal barrier. The expression of these three genes in the DIO-P group was significantly lower than that in the DIO-R group (P0.05), indicating that the intestinal barrier might be damaged in the obese susceptible group. Elisa analysis showed that the blood pro-inflammatory cytokines in DIO-P group increased significantly (P0.05), while the content of anti-inflammatory cytokines decreased significantly (P0.05), indicating the occurrence of chronic systemic inflammation. In conclusion, this study shows that the susceptibility to obesity in mice under high fat diet is closely related to intestinal flora. The mechanism of intestinal flora affecting obesity susceptibility may be related to injury of intestinal mucosal barrier, inflammatory response and disturbance of lipid metabolism.
【學(xué)位授予單位】：江南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R589.2
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