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標題: | 短鏈脂肪酸與嬰兒異位性皮膚炎及高脂飲食小鼠腸道免疫之相關性探討 Study on the association of short chain fatty acids with infant atopic dermatitis and intestinal immunity of high-fat diet fed mice |
作者: | Ying-Rou Shen 沈瑩柔 |
指導教授: | 林璧鳳 |
關鍵字: | 異位性皮膚炎,脂肪酸,短鏈脂肪酸,母乳,腸道免疫,比菲德氏菌,益生菌,寒天寡醣, atopic dermatitis,fatty acids,short chain fatty acids,breast milk,intestinal immunity,Bifidobacterium,probiotics,agaro-oligosaccharide, |
出版年 : | 2017 |
學位: | 碩士 |
摘要: | 短鏈脂肪酸為腸道菌代謝水溶性纖維的產物,與腸道免疫的關係日益受到重視,目前被認為具有抗發炎的作用,能改善氣喘、肥胖、腸道發炎等疾病。故本研究探討短鏈脂肪酸是否與過敏疾病和腸道免疫反應有關。分別檢測異位性皮膚炎嬰兒所哺餵的母乳,及給予高油飲食小鼠添加益生質寒天寡醣,探討短鏈脂肪酸與免疫反應的相關性。第一部份實驗樣品於臺大醫院小兒科過敏門診,收集6個月齡以下罹患異位性皮膚炎嬰兒的母乳,分別測定短鏈脂肪酸 (C2~C4) 及中長鏈脂肪酸 (C8-C24) 含量,並對母親進行1個月的飲食頻率問卷調查。結果顯示,異位性皮膚炎嬰兒所攝取母乳的MUFAs、n-3 PUFAs、palmitic acid及arachidonic acid脂肪酸百分比較高,而capric acid和myristic acid較低,短鏈脂肪酸則無顯著差異。第二部分以高油飲食小鼠模式,探討短鏈脂肪酸和腸道菌組成對腸道免疫之影響。實驗設計為給予5週齡C57BL/6J小鼠28% 高油飼料 (HF組,油脂熱量為51%),高油添加3%寒天寡醣 (HF-AOS組)、或添加109/day CFU市售益生菌粉末 (HF-PRO組)。餵飼8週後犧牲,分析脾臟、腸繫膜淋巴結與皮耶式體的免疫指標,盲腸和糞便的短鏈脂肪酸以及糞便IgA含量。結果顯示,高油飲食造成小鼠糞便比菲德氏菌屬顯著下降,而HF-PRO組添加益生菌確實顯著增加糞便比菲德氏菌屬的含量。HF-PRO組小鼠MLN的CD103+樹突細胞比例也較高,PP細胞分泌IL-2顯著較HF組低,脾臟細胞和MLN細胞IL-17分泌量趨勢較HF組高,而脾臟細胞IL-2分泌量顯著較HF組高。HF-AOS組母鼠宮旁脂顯著最輕,血清和肝臟脂質含量顯著最少,但在糞便中顯著最多,顯示寒天寡醣可能會抑制脂質吸收。HF-AOS組盲腸顯著最重,糞便的腸桿菌科含量略高,盲腸及糞便的短鏈脂肪酸含量顯著最低。MLN細胞分泌IFN-γ有增加的趨勢,糞便IgA的濃度顯著最高。另外大腸組織切片也觀察到絨毛破損、黏液分泌量較少,推測本實驗添加寒天寡醣的量偏高可能吸附腸道水份,造成黏膜細胞受損,腸道蠕動較快速,使得腸道菌無法利用寒天寡醣發酵生成短鏈脂肪酸,使腸道免疫趨向抵抗感染的發炎反應。綜合以上,本研究發現母乳有較低比率的SFAs、較高比率的MUFAs及n-3 PUFAs可能與嬰兒異位性皮膚炎相關,但未呈現母乳的短鏈脂肪酸含量與嬰兒異位性皮膚炎有關。高油飲食小鼠添加寒天寡醣劑量可能過高,導致小鼠輕微腹瀉且腸道處於較發炎的狀態,而高油飲食添加益生菌,顯著降低PP細胞IL-2的分泌量,但有促進MLN細胞分泌IL-17及脾臟細胞分泌IL-2和IL-17的趨勢,意喻高油飲食下補充益生菌,改善腸道免疫的效果有限。 The microbial metabolites short chain fatty acids (SCFAs) are fermented from soluble dietary fiber. Significant attention has been focused on the relationship between SCFAs and intestinal immunity. SCFAs was shown to have the anti-inflammatory effect, and could ameliorate asthma, obesity, intestinal inflammation. Thus, the aim of this study is to investigate the association of SCFAs with allergic disease and intestinal immunity. To investigate the effect of SCFAs on immune response, we analyzed breast milk from the infant with atopic dermatitis (AD), and related parameters from high-fat diet fed mice supplemented with prebiotics agaro-oligosaccharide. In the first part, we cooperated with the Department of the Pediatric at NTU hospital to collect breast milk from infants’ mothers to detect the C8-C24 fatty acids composition and short chain fatty acids (SCFAs) concentration. We also analyzed maternal diet by FFQ (food frequency questionnaire). The results showed that the percentages of MUFAs, n-3 PUFAs, palmitic acid and arachidonic acid of breast milk were higher, and capric acid and myristic acid were lower in the AD group than the health group. SCFAs concentration had no significant difference between two groups. In the second part, we investigated the effect of SCFAs and microbiota on intestinal immunity in high-fat diet fed mice. 5 week-old C57BL/6J mice were fed either 28% fat diet (HF group), HF containing 3% AOS (HF-AOS group) or 109/day CFU probiotics (HF-PRO group) for 8 weeks. Immune indicators of spleen, MLN, PP, fecal and cecal SCFAs, and fecal IgA were also detected. The results showed that fecal Bifidobacterium was decreased in high-fat diet fed mice, and probiotics increased fecal Bifidobacterium in the HF-PRO group. CD103+ dendritic cell percentage was increased in MLN, IL-2 secretion from PP was lower, IL-17 secretion from MLN was higher, and IL-2 and IL-17 secretion from splenocytes were higher in the HF-PRO group compared with the HF group. The parametrial white adipose from female mice was the lightest in the HF-AOS group. The lipid content in serum and liver were the least, but the lipid content was the highest in the HF-AOS group. These data showed that AOS might inhibit the absorption of lipid. The cecum was heaviest, fecal Enterobacteriaceae increased, and fecal and cecal SCFAs were decreased in the HF-AOS group. Fecal IgA and IFN-γ secreted from MLN cells were increased in the HF-AOS group. Evaluating colon histomorphology from H&E and PAS stained, the villous were damaged and with less mucin. We surmised that the mucus was damaged due to the high dose of AOS and its water containing ability. It also led to pro-inflammatory response in mice intestine with less SCFAs production. In conclusion, the lower percentage of SFAs and the higher percentage of MUFAs and n-3 PUFAs might be associated with infant AD. But there was no association between breast milk SCFAs and infant AD. Our results found out that 3% AOS in high-fat diet fed mice might lead to dysbiosis and inflammation in mice intestine, which might due to overdose. High-fat diet with probiotics could decrease IL-2 secretion from PP, but tend to promote IL-17 secretion in MLN and IL-2 and IL-17 secretion in spleen. Thus, the results indicated that the effect of probiotics on high-fat diet fed mice was limited. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/69066 |
DOI: | 10.6342/NTU201703241 |
全文授權: | 有償授權 |
顯示於系所單位: | 生化科技學系 |
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