【摘要】： 背景：益生菌對健康的積極作用越來越受到人們的重視。乳酸桿菌作為目前臨床應(yīng)用最為廣泛的益生菌之一，其在IBD、IBS、慢性腹瀉等有腸道炎癥基礎(chǔ)的疾病治療中的重大價值已經(jīng)被許多高水平的臨床研究證實。然而關(guān)于益生菌對腸道炎癥作用機制方面的研究雖然開展多年，但是對此問題仍沒有統(tǒng)一全面的認識。腸上皮細胞和腸粘膜固有層免疫細胞作為腸道屏障的重要組成均有可能參與益生菌對腸道炎癥的調(diào)控，它們各自在腸道炎癥中的作用已被一些研究部分證實，但缺乏有效的研究比較兩者在益生菌調(diào)控腸道炎癥方面的作用。細胞因子TNF-α幾乎存在于所有的炎癥反應(yīng)當(dāng)中，它的強大的促炎癥作用能夠激活炎癥介質(zhì)聯(lián)級反應(yīng)并動員炎癥細胞。測定TNF-α是評價腸道炎癥的理想指標(biāo)。 目的：研究乳酸桿菌(Lactobacillus casei CICC6002)對腸上皮細胞(HT-29)和單核細胞(THP-1)TNF-α分泌的影響，并對照非致病性大腸埃希菌(Escherichia coli DH5α)對細胞TNF-α分泌的影響情況，探索益生菌的抗炎作用及其作用途徑。 方法：將37℃有氧條件下，MRS、LB細菌培養(yǎng)基中處于對數(shù)生長期的L casei、E coli各自轉(zhuǎn)種于RPMI培養(yǎng)液孵育48小時，以100 000g×1小時，，4℃超速離心收集含細菌分泌代謝產(chǎn)物的條件培養(yǎng)基(CM)；培養(yǎng)腸上皮細胞(HT-29)和單核細胞(THP-1)；以脂多糖(LPS)刺激HT-29細胞和THP-1細胞分泌TNF-α，加入細菌的CM，共孵育4小時。以ELISA的方法檢測L casei CM、E coli CM對HT-29細胞、THP-1細胞TNF-α分泌的影響。數(shù)據(jù)進行t檢驗。 結(jié)果：LPS刺激，加或不加細菌CM[956.39±374.45，475.74±49.59，829.62±110.51pg/ml]，均使THP-1細胞TNF-α的分泌比基礎(chǔ)狀態(tài)[100.96±48.95pg/ml]明顯增加(P＜0.05)。與單純LPS刺激的THP-1細胞TNF-α分泌[956.39±374.45pg/ml]相比，加入L casei CM顯著減少LPS刺激的THP-1細胞TNF-α分泌[475.74±49.59pg/ml](P＜0.05)；加入E coli CM，LPS刺激的THP-1細胞TNF-α分泌[829.62±110.51pg/ml]無明顯變化(P＞0.05)。不加任何刺激、單純LPS刺激或者LPS與L casei CM聯(lián)合刺激條件下均未能檢出HT-29細胞TNF-α分泌。LPS與E coli CM聯(lián)合刺激可使HT-29細胞產(chǎn)生TNF-α[10.94±6.32pg/ml]，明顯低于相同條件下THP-1細胞TNF-α分泌[829.62±110.51pg/ml](P＜0.01)。 結(jié)論：益生菌干酪乳酸桿菌具有抗炎特性，對抑制腸道炎癥意義重大。干酪乳酸桿菌抗炎特性通過減少單核細胞分泌TNF-α實現(xiàn)。腸上皮細胞沒有參與干酪乳酸桿菌的抗炎過程。非致病性大腸埃希菌DH5α無抗炎特性。
[Abstract]:Background: people pay more and more attention to the positive effect of probiotics on health. Lactobacillus is one of the most widely used probiotics in clinic at present. Its important value in the treatment of IBD,IBS, chronic diarrhea and other diseases based on intestinal inflammation has been confirmed by many high levels of clinical research. However, although the studies on the mechanism of probiotics on intestinal inflammation have been carried out for many years, there is still no unified and comprehensive understanding of this problem. As an important component of intestinal barrier, both intestinal epithelial cells and intestinal mucosal lamina immune cells may be involved in the regulation of intestinal inflammation by probiotics. Their respective roles in intestinal inflammation have been partly confirmed by some studies. But there is a lack of effective research to compare the role of probiotics in the regulation of intestinal inflammation. Cytokine TNF- 偽 exists in almost all inflammatory responses, its powerful pro-inflammatory role can activate the inflammatory mediators of the chain reaction and mobilize inflammatory cells. Determination of TNF- 偽 is an ideal index for evaluating intestinal inflammation. Objective: to study the effect of Lactobacillus (Lactobacillus casei CICC6002 on the secretion of TNF- 偽 in intestinal epithelial cells (HT-29) and monocytes (THP-1), and to explore the anti-inflammatory effect of probiotics and its pathways by comparing the effect of non-pathogenic Escherichia coli (Escherichia coli DH5 偽 on the secretion of TNF- 偽. Methods: l casei,E coli, which is in logarithmic growth phase, was incubated in RPMI medium at 37 鈩

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