近日，華中農(nóng)業(yè)大學(xué)魏泓教授團(tuán)隊針對腸道微生物群體感應(yīng)（QS）信號分子調(diào)控宮內(nèi)生長受限（IUGR）仔豬腸道損傷這一領(lǐng)域開展系統(tǒng)深入的研究，在國際學(xué)術(shù)期刊Science of the Total Environment，Gut Microbes，Microbiology Spectrum，F(xiàn)rontiers in Immunology，Journal of Cellular Physiology和The Journal of Nutrition上發(fā)表了系列研究論文。揭示了IUGR仔豬腸腔中微生物QS信號分子的變化及其調(diào)控腸道屏障功能損傷的分子機制，為預(yù)防IUGR所致腸道疾病的發(fā)生提供了新思路，為IUGR仔豬營養(yǎng)生理調(diào)控關(guān)鍵靶點的篩選提供了理論依據(jù)。   　　IUGR導(dǎo)致的仔豬低初生重及繼發(fā)的哺乳期高死亡率和全期低生長性能，是養(yǎng)豬業(yè)存在的主要問題之一。據(jù)統(tǒng)計，受IUGR影響，我國15%~20%的仔豬初生重低于1.1 kg。IUGR仔豬通常伴有出生后腸道功能障礙，從而影響動物出生后的生產(chǎn)潛力。相比于正常仔豬，IUGR仔豬的后期飼料利用效率降低30%，平均出欄時間延長30天，每年給我國養(yǎng)豬生產(chǎn)帶來的經(jīng)濟損失達(dá)150億元。因此，改善IUGR仔豬的腸道健康狀況對于確保生豬健康養(yǎng)殖十分重要而迫切。研究團(tuán)隊前期發(fā)現(xiàn)IUGR所致的腸道發(fā)育缺陷可以從新生持續(xù)到生長階段，由氧化應(yīng)激和細(xì)胞凋亡所致的腸道屏障功能損傷是IUGR豬腸道發(fā)育缺陷的生物學(xué)基礎(chǔ)。   　　QS是原核生物中一種基于分子信號的通訊機制。QS信號分子不僅能夠控制微生物群落行為，還可調(diào)節(jié)宿主細(xì)胞的生理狀態(tài)。研究表明，革蘭氏陰性菌通過分泌QS信號分子調(diào)節(jié)真核細(xì)胞的功能，破壞宿主腸道上皮細(xì)胞的穩(wěn)態(tài)，最終導(dǎo)致腸上皮屏障功能障礙。研究團(tuán)隊前期發(fā)現(xiàn)，以擬桿菌屬和梭桿菌屬為代表的革蘭氏陰性菌是IUGR仔豬腸道中的優(yōu)勢菌群。然而，目前尚不清楚IUGR仔豬腸腔內(nèi)革蘭氏陰性菌的大量繁殖是否增加QS信號分子的分泌？基于此科學(xué)問題，通過檢測正常和IUGR仔豬糞便中9種典型的革蘭氏陰性菌源的QS信號分子濃度發(fā)現(xiàn)3OC12-HSL在IUGR仔豬中顯著升高；以瘤胃球菌為代表的革蘭氏陽性菌在IUGR仔豬腸道中顯著降低，且?guī)缀跛械牟町惲鑫盖蚓c3OC12-HSL濃度呈顯著負(fù)相關(guān)，該結(jié)果為IUGR仔豬腸腔內(nèi)存在革蘭氏陰性菌來源的QS信號分子提供了直接證據(jù)。   　　研究剖析了微生物QS信號分子3OC12-HSL介導(dǎo)IUGR仔豬腸道屏障功能損傷的分子機制。體外細(xì)胞試驗結(jié)果表明：3OC12-HSL可通過誘導(dǎo)腸上皮細(xì)胞氧化應(yīng)激與凋亡、破壞細(xì)胞外基質(zhì)與緊密連接蛋白，從而影響腸上皮細(xì)胞增殖、破壞屏障功能；而抗氧化劑NAC和細(xì)胞凋亡抑制劑Z-VAD-FMK可阻止3OC12-HSL對腸上皮屏障的破壞作用，進(jìn)一步明確了氧化應(yīng)激與凋亡通路在該過程中的介導(dǎo)作用。基于無菌小鼠體系進(jìn)一步揭示了3OC12-HSL導(dǎo)致腸道屏障功能損傷的微生物介導(dǎo)機制：3OC12-HSL破壞SPF小鼠腸道上皮屏障，誘導(dǎo)全身炎癥反應(yīng)；將SPF小鼠的糞便微生物移植給無菌小鼠后，無菌小鼠幾乎復(fù)制了3OC12-HSL處理的SPF小鼠的所有表型；通過微生物組學(xué)聯(lián)合分析發(fā)現(xiàn)Elizabethkingia spp.是SPF和無菌小鼠腸道中共同的差異細(xì)菌，該研究利用無菌動物模型和糞便微生物移植技術(shù)，首次在動物體內(nèi)證明了QS信號分子3OC12-HSL通過調(diào)控特定微生物破壞腸道屏障功能，該研究方法為解析豬功能微生物與宿主互作機制提供了新思路。基于上述研究成果形成了綜述性論文，該論文對革蘭氏陰性菌QS信號分子在調(diào)節(jié)宿主細(xì)胞功能和腸道健康中的重要性進(jìn)行了全面的綜述，并提出通過阻斷QS信號分子發(fā)揮其功能的途徑來應(yīng)用于防治人類和動物腸道疾病的可能性。   　　該研究團(tuán)隊長期致力于腸道功能微生物的發(fā)掘及利用無菌動物體系對功能微生物進(jìn)行功能驗證。通過整合腸道功能微生物組與無菌動物實驗技術(shù)，發(fā)現(xiàn)了糞菌移植通過調(diào)控宿主免疫和腸道屏障功能緩解無菌小鼠結(jié)腸炎的敏感性；闡明了益生菌（丁酸梭菌）及其衍生的胞外囊泡通過調(diào)節(jié)腸道內(nèi)穩(wěn)態(tài)及腸道菌群以改善急性實驗性結(jié)腸炎，該研究為以腸道功能微生物為靶點防治炎癥性腸道疾病提供了新的見解，并促進(jìn)了炎癥性腸道疾病的新型治療和預(yù)防干預(yù)措施的發(fā)展。   　　華中農(nóng)業(yè)大學(xué)魏泓教授團(tuán)隊陶詩煜副研究員為系列論文的第一或通訊（含共同）作者，研究得到中國農(nóng)業(yè)大學(xué)王軍軍教授和浙江省農(nóng)業(yè)科學(xué)院楊華研究員的支持，上述研究受到國家自然科學(xué)基金、國家重點研發(fā)計劃和校自主科技創(chuàng)新基金的資助。   　　【英文摘要1】   　　Quorum sensing is a molecular signaling-based communication mechanism in prokaryotes. In the basic mode, signaling molecules released by certain bacteria are sensed by intracellular receptors or membrane-bound receptors of other members in the community, leading to the collective isogenic signaling molecule synthesis and synchronized activities. This regulation is important for the symbiosis of the bacterium with the host, as well as virulence and biofilm formation. Notably, quorum sensing signaling molecules are not only able to control microbial community behavior but can likewise regulate the physiological status of host cells. Here, we provide a comprehensive review of the importance of quorum sensing signaling molecules in gram-negative bacteria in regulating host cell function and gut health, and suggest possible opportunities for application in combating human and animal diseases by blocking the pathways through which quorum sensing signaling molecules exert their functions.   　　【英文摘要2】   　　As a quorum sensing signal molecule, N-（3-oxododecanoyl）-homoserine lactone （3OC12） regulate the population behavior of microorganisms. Many studies have proved that 3OC12 harm the physiological function of host intestinal epithelial cells. However, the detrimental effects of 3OC12 on intestinal health need verification in animals. Besides, the role of gut microbiome in 3OC12-induced intestinal damage also needs further understanding. In our study, 3OC12 was first administered to specific pathogen-free （SPF） mice, then the fecal microbiome of SPF mice was transplanted into germ-free （GF） mice to reveal the effects of 3OC12 on intestinal health and regulatory mechanisms of the intestinal microbiome. 3OC12 treatment significantly decreased body weight, shortened colonic length, disrupted the morphology of the colonic epithelium and increased the histopathological score of the colon in SPF mice. The levels of diamine peroxidase, d-lactate, TNF-α， IL-1β， and IL-8 were found to be significantly elevated in the serum of 3OC12 mice, while the levels of IL-10 were significantly reduced. Besides, the fecal microbial community of mice was also altered in the 3OC12-treated SPF mice. The results of fecal microbial transplantation （FMT） experiment showed that the phenotypes in SPF mice were almost reproduced in GF mice, manifested by body weight loss, colon damage and changed in serum chemical markers. More importantly, a joint analysis of fecal microbes in SPF and GF mice revealed Feature14_Elizabethkingia spp. was common differential bacteria in the feces of two kinds of mice treated with and without FMT. Our results demonstrated that 3OC12 challenge led to systemic inflammation and body weight loss in mice by disrupting intestinal barrier function, in which gut microbiome played a key role. These findings increased our understanding of the mechanism of intestinal injury caused by 3CO12, providing new ideas for the prevention and therapy of diseases caused by bacterial infection from the perspective of intestinal microbiome.   　　【英文摘要3】   　　Microbes employ autoinducers of quorum sensing （QS） for population communication. Although the autoinducer of Pseudomonas aeruginosa LasI-LasR system, N-（3-oxododecanoyl）- l -homoserine lactone （3OC12）， has been reported with deleterious effects on host cells, its biological effects on integrity of the intestinal epithelium and epithelial barrier are still unclear and need further investigation. In the present study, flow cytometry, transcriptome analysis and western blot technology have been adopted to investigate the potential molecular mechanisms of 3OC12 and its structurally similar analogs damage to intestinal epithelial cells. Our results indicated that 3OC12 and 3OC14 trigger apoptosis rather than necrosis and ferroptosis in intestinal epithelial cells. RNA-sequencing combined with bioinformatics analysis showed that 3OC12 and 3OC14 reduced the expression of genes from extracellular matrix （ECM）-receptor interaction pathway. Consistently, protein expressions from ECM and tight junction-associated pathway were significantly reduced after 3OC12 and 3OC14 challenge. In addition, 3OC12 and 3OC14 led to blocked cell cycle, decreased mitochondrial membrane potential, increased reactive oxygen species level and elevated Ca2+ concentration. Reversely, the antioxidant NAC could effectively mitigate the reduced expression of ECM and tight junction proteins caused by 3OC12 and 3OC14 challenge. Collectively, this study demonstrated that QS autoinducer exposure to intestinal epithelial cells ablates the ECM and tight junctions by triggering oxidative stress and apoptosis, and finally disrupts the intestinal epithelial barrier. These findings provide a rationale for defensing QS-dependent bacterial infections and potential role of NAC for alleviating the syndrome.   　　【英文摘要4】   　　Microbiological treatments are expected to have a role in the future management of inflammatory bowel disease （IBD）。 Clostridium butyricum （C. butyricum） is a probiotic microorganism that exhibits beneficial effects on various disease conditions. Although many studies have revealed that C. butyricum provides protective effects in mice with colitis, the way C. butyricum establishes beneficial results in the host remains unclear. In this study, we investigated the mechanisms by which C. butyricum modifies the gut microbiota, produces bacterial metabolites that may be involved, and, specifically, how microbial extracellular vesicles （EVs） positively influence IBD, using a dextran sulfate sodium （DSS）-induced colitis murine model in mice. First, we showed that C. butyricum provides a protective effect against colitis, as evidenced by the prevention of body weight loss, a reduction in the disease activity index （DAI） score, a shortened colon length, decreased histology score, and an improved gut barrier function, accompanied by reduced levels of pathogenic bacteria, including Escherichia/Shigella, and an increased relative abundance of butyrate-producing Clostridium sensu stricto-1 and Butyricicoccus. Second, we also confirmed that the gut microbiota and metabolites produced by C. butyricum played key roles in the attenuation of DSS-induced experimental colitis, as supported by the profound alleviation of colitis effects following fecal transplantation or fecal filtrate insertion supplied from C. butyricum-treated mice. Finally, C. butyricum-derived EVs protected the gut barrier function, improved gut microbiota homeostasis in ulcerative colitis, and contributed to overall colitis alleviation.   　　論文鏈接：   　　https://www.tandfonline.com/doi/full/10.1080/19490976.2022.2039048   　　https://www.sciencedirect.com/science/article/pii/S0048969721014157?via%3Dihub   　　https://onlinelibrary.wiley.com/doi/10.1002/jcp.30261   　　https://academic.oup.com/jn/article/151/7/1736/6274958?login=true   　　https://journals.asm.org/doi/full/10.1128/spectrum.01368-22?rfr_dat=cr_pub++0pubmed&url_ver=Z39.88-2003&rfr_id=ori%3Arid%3Acrossref.org   　　https://www.frontiersin.org/articles/10.3389/fimmu.2022.836542/full