山银花对小鼠肠道菌群失衡的调节作用

目的探讨山银花对于小鼠肠道菌群失衡的调节作用。方法应用头孢曲松钠灌胃的方法建立小鼠肠道菌群失衡模型。利用观察称量等方法观察小鼠临床表征的变化,以及盲肠指数和脾指数的变化。利用活菌计数法分析肠道优势菌群的变化趋势。结果 (1)与正常对照组相比,菌群失衡组盲肠指数增高(P<0.05);自然恢复组变化不明显;山银花组(5%、10%)与大豆低聚糖组相似,显著下降(P<0.05),呈恢复趋势。(2)菌群失衡组脾脏指数有下降趋势,自然恢复组无明显变化;山银花组(5%、10%)与大豆低聚糖组均有所上升,呈恢复趋势。(3)菌群失衡组厌氧菌被抑制,需氧菌大量繁殖,自然恢复组菌群种类和数量变化不明显,主要是需氧菌的恢复;山银花组(5%、10%)和大豆低聚糖组厌氧菌重新出现,菌群种类增多,尤其是双歧杆菌、乳杆菌、韦荣球菌和类杆菌的出现预示着菌群正在逐渐恢复正常。10%山银花高剂量组比5%山银花低剂量组恢复要好。结论山银花可以促进菌群失衡小鼠盲肠和脾脏大小的恢复,可以调节小鼠肠道菌群失衡,具有益生元功效。     

口炎清颗粒对小鼠肠道菌群失衡的调节作用

目的探讨口炎清颗粒对肠道菌群失衡的调节作用。方法小鼠随机分为6组：正常对照组、重度失调组、自然恢复组、口炎清低剂量组、口炎清高剂量组和大豆低聚糖对照组。正常对照组小鼠灌服蒸馏水,其他组以头孢曲松钠终浓度为8 g/（kg.d）的剂量,连续灌胃8 d,建立小鼠菌群失衡模型,自然恢复组在失衡模型建立后不使用任何药物,自然恢复7 d;口炎清低剂量组、高剂量组、大豆低聚糖对照组分别灌服相应剂量的药物7 d,观察各组小鼠盲肠指数和肠道内双歧杆菌、类杆菌、韦荣球菌、乳酸杆菌、消化球菌、优杆菌、葡萄球菌、肠球菌、大肠埃希菌、链球菌和酵母菌11种菌群变化。结果与菌群失衡组比较,口炎清组与大豆低聚糖组盲肠指数均显著下降（P〈0.05）,呈恢复趋势。自然恢复组肠道菌群种类和数量变化不明显,口炎清组厌氧菌重新出现,尤其是低剂量组双歧杆菌、优杆菌和类杆菌的出现预示着肠道菌群正在逐渐恢复正常。结论一定浓度的口炎清可以明显促进菌群失衡小鼠盲肠大小的恢复,对小鼠肠道菌群有调节作用,具有类似大豆低聚糖的益生元功效。     

枸杞子提取物对小鼠肠道菌群失衡的调整作用

本研究旨在探讨枸杞子不同浓度提取物对小鼠肠道菌群失衡的调整作用,为枸杞子的进一步开发利用提供理论依据。使用抗生素建立肠道菌群失衡模型,将小鼠随机分为正常对照组、枸杞子提取物高浓度组、低浓度组、模型组和自然恢复组,以双歧杆菌、乳酸杆菌、肠杆菌、肠球菌等肠道正常菌群为对象,利用传统菌落计数法研究各组肠道菌群失衡及恢复调整情况。枸杞子提取物对乳酸杆菌和双歧杆菌的生长具有恢复作用,且低浓度提取物的增殖作用优于高浓度,而对肠球菌和肠杆菌的作用并不明显。枸杞子提取物对小鼠肠道菌群失衡有一定的调整作用,能够纠正抗生素相关性菌群失调。     

小鼠肠道菌群失衡模型建立

目的利用抗生素干扰小鼠肠道菌群建立轻重程度的菌群失衡小鼠模型。方法选用不同浓度的头孢曲松钠,行小鼠灌胃,取盲肠内容物连续观察培养优势菌群变化。结果优势菌群失衡组小鼠与对照比较,盲肠体积增大,盲肠指数升高。头孢曲松钠8g/(kg·d)剂量连续灌胃8d,造成重度失衡模型,小鼠肠道只能检出肠杆菌属、链球菌属,且数量被抑制在103CFU/g以下。头孢曲松钠5g/(kg·d)剂量连续灌胃8d,造成轻度失衡模型,小鼠肠道双歧杆菌属、链球菌属和韦荣球菌属数量降至105左右,类杆菌属降至104左右。消化球菌属降至103左右,而其他菌属如乳酸杆菌属、肠球菌属和肠肝菌属等数量显著降低。结论亚致死剂量头孢曲松钠灌胃可以建立肠道菌群重度失衡模型。     

香菇多糖对微生态失调小鼠肠道菌群及免疫功能的调节作用

目的 探讨香菇多糖对微生态失调小鼠肠道菌群及免疫功能的调节作用.方法 经盐酸林可霉素灌胃建立肠道微生态失调小鼠模型,香菇多糖灌胃治疗,同时设正常对照组、自然恢复组和丽珠肠乐组.7d后处死各组小鼠,进行肠道菌群定量、免疫器官体重及其淋巴细胞转化率检测.结果 用香菇多糖对肠道微生态失调小鼠进行治疗后,小鼠肠道双歧杆菌、乳酸杆菌数量显著增加,而肠杆菌和肠球菌的数量显著降低;脾脏指数明显增加,对胸腺指数无影响;显著增强了淋巴细胞转化率.结论 盐酸林克霉素灌胃能诱导微生态失调小鼠模型的有效建立.香菇多糖能调整小鼠肠道菌群及免疫功能.     

豹皮樟提取物对小鼠肠道菌群失衡的调整作用

目的观察豹皮樟（老鹰茶）不同溶剂提取物对小鼠抗生素相关性菌群失衡的调整作用,为豹皮樟的进一步开发利用提供理论依据。方法使用抗生素造模,将小鼠分为正常对照组、水提物组、醇提物组、模型组和自然恢复组,利用传统菌落计数法等观察各组肠道菌群失衡及恢复调整情况。结果豹皮樟不同提取物对改善小鼠肠道内菌群的效果不同：水提物具有显著调节肠杆菌及肠球菌等小鼠肠道表层需氧菌的作用,醇提物对小鼠肠道深层厌氧双歧杆菌和乳酸杆菌的调整作用显著,具有益生元特眭。结论豹皮樟可以调整肠道菌群,纠正抗生素相关性菌群失调。     

中药"神曲"对脾虚小鼠肠道菌群的调整及肠保护作用研究

目的研究中药"神曲"对脾虚小鼠肠道菌群的调整及肠保护作用.方法1.昆明小白鼠,分成4组:A组(正常对照组),B组(脾虚自然恢复组),C组(神曲治疗组),D组(丽珠肠乐治疗组).2.检测结肠内容物和粪便标本的肠杆菌、肠球菌、双歧杆菌、类杆菌和乳酸杆菌数量.3.结肠组织光镜和电镜观察.结果脾虚小鼠给予神曲水煎剂治疗后,肠杆菌、肠球菌、双歧杆菌、类杆菌和乳酸杆菌数量逐渐恢复正常,肠壁肌层厚度增加,杯状细胞数量增多,肠黏膜微绒毛排列紊乱、线粒体肿胀显著改善,明显好于自然恢复组,与丽珠肠乐治疗组相比差异无显著性.结论中药神曲对脾虚小鼠具有肠道菌群调整及促进损伤肠组织恢复的作用.     

大豆低聚糖对肠道菌群结构调节的研究

目的 :对山松牌大豆低聚糖对肠道菌群调节的效果进行研究。方法 :通过动物实验和人体试验。结果 :山松牌大豆低聚糖能增殖体内乳杆菌和双歧杆菌数量 ;在较高剂量时 ,能使体内的肠球菌和肠杆菌增殖。采用B/E值 (即双歧杆菌和肠杆菌的比值 )作为指标进行分析发现 ,该制品在低剂量时能更好地优化肠道菌群结构。结论 :大豆低聚糖对肠道菌有较好的调节效果     

模拟失重对大鼠肠道菌群影响的研究

目的：研究模拟失重条件下大鼠肠道菌群变化情况。方法：选择性培养基分别对肠球菌、肠杆菌、类杆菌、乳杆菌以及双歧杆菌进行定量测定,扫描电镜观察大鼠盲肠上皮细胞的组织变化。结果：过路菌群中肠杆菌和肠球菌数量增加显著;原籍菌群中双歧杆菌减少十分明显,乳杆菌的数量也有不同程度的减少,而类杆菌的数量有增加的趋势。SD大鼠盲肠出现了肿胀细胞,上皮细胞颈毛排列紊乱、稀疏。结论：模拟失重条件下大鼠肠道微生态出现平衡失调。     

全科广谱治疗仪对由抗生素导致小鼠肠道菌群失调的调整作用

用盐酸林可霉素导致小鼠肠道菌群失调,然后用ＱＫ－ ＣＯＩＣ 型全科广谱治疗仪进行照射,并分别于４ｄ和７ｄ 采取照射组与自然恢复组小鼠粪便,对肠杆菌（ＥＭＢ） 、肠球菌（Ｅｃ） 、乳杆菌（Ｌｂｓ） 、双歧杆菌（ＢＳ） 和类杆菌（Ｂｄｂ）进行菌群分析。结果照射组４ｄ 菌群便有所恢复,７ｄ 时已基本恢复正常,与自然恢复组比有显著差异（ｐ ＜ ０．０５） 。     

长链菊粉对抗生素处理后小鼠肠道菌群重建的影响

【目的】评价长链菊粉对抗生素致小鼠肠道菌群失调后肠道菌群的恢复情况。【方法】选择50只健康的10周龄BALB/c小鼠,随机分为2组,其中15只为正常对照组,余下35只饮用水中含4种抗生素连续喂养7 d,诱导小鼠肠道菌群严重失调后,再随机分为长链菊粉恢复组(饮用水中添加5%(W/W)长链菊粉)和自发恢复组(饮用水为无菌水),连续处理21 d。受试小鼠在抗生素治疗后的第7天以及恢复喂养的第7、14和21天,取结肠组织进行切片然后进行HE染色分析,无菌取粪便进行16Sr RNA测序分析,观察小鼠肠道组织及菌群恢复情况。【结果】抗生素处理7 d后,小鼠结肠组织有轻微炎症,但肠道菌群严重失调。组织学分析表明,在补充长链菊粉或自发恢复21d后,结肠炎症逐渐减轻;但相比于自发恢复,长链菊粉干预延迟了结肠组织的恢复。16S r RNA基因V3–V4区扩增子测序分析显示无论是长链菊粉补充还是自发恢复都无法在属水平上恢复肠道菌群组成。尤其是长链菊粉的补充,反而导致了某些机会致病菌的选择性扩增,并提高了与肠道菌群相关的疾病途径。【结论】抗生素诱导肠道菌群严重失调后补充长链菊粉会延迟肠道菌群的重建,可能会导致潜在的不良影响。     

高脂食物对动脉硬化合并类风湿关节炎 小鼠肠道菌群的影响

目的分析高脂食物对动脉硬化合并类风湿关节炎小鼠肠道微生物的影响,了解动脉硬化合并类风湿关节炎小鼠肠道微生物的变化。方法 8周龄ApoE~(-/-)小鼠饲喂高脂食物和普通食物至17周龄来诱发动脉硬化症状,再通过给17周龄ApoE~(-/-)小鼠腹腔注射抗6-磷酸葡萄糖异构酶(glucose-6-phosphate isomerase,GPI)抗体呈阳性的K/BxN血清,从而诱导其产生类风湿关节炎症状。通过Illumina HiSeq平台对各组小鼠粪便进行16S rDNA V4区测序,分析动脉硬化合并类风湿关节炎小鼠肠道微生物的变化。结果 ApoE~(-/-)小鼠饲喂高脂食物后,其血清低密度脂蛋白胆固醇(LDL-C)浓度和血清总胆固醇(TC)浓度均显著升高,主动脉内膜斑块面积比喂普通食物的ApoE~(-/-)小鼠显著增加,表明ApoE~(-/-)小鼠饲喂高脂食物后引起更显著的动脉硬化症状。再通过腹腔注射抗GPI抗体呈阳性的K/BxN血清,各组ApoE~(-/-)小鼠均出现关节肿胀,饲喂高脂食物的ApoE~(-/-)小鼠其踝关节宽度和临床评分(clinical score)低于饲喂普通食物组小鼠。OTU数、Shannon指数和Simpson指数显示高脂食物和K/BxN血清处理组ApoE~(-/-)小鼠肠道菌群多样性降低,Firmicutes/Bacteroidetes值升高,t-test分析显示在属水平上,Prevotellaceae_UCG-001显著降低,Ruminiclostridium_6显著升高。t-test分析和Firmicutes/Bacteroidetes比值显示ApoE~(-/-)小鼠肠道菌群结构紊乱。结论高脂食物使ApoE~(-/-)小鼠的肠道菌群组成和结构发生改变,导致ApoE~(-/-)小鼠的动脉硬化症状加重,类风湿关节炎症状减轻。提示肠道微生物组成和结构的改变,可能与动脉硬化合并类风湿关节炎发病机制相关。     

马齿苋多糖对肠道微生态失调小鼠的调整作用研究

目的研究应用马齿苋多糖对肠道微生态失调小鼠进行调整治疗,达到从微生态学角度防治感染的目的。方法应用林可霉素灌胃建立肠道微生态失调小鼠模型,然后用马齿苋多糖进行治疗,同时设正常对照组、阳性对照组和阴性对照组,于给药7 d后处死小鼠,进行肠道菌群定量、肠内容物挥发性脂肪酸检测及肠黏膜电镜观察。结果林可霉素灌胃3 d后,小鼠肠道菌群失调,肠内容物挥发性脂肪酸含量明显下降,肠黏膜损伤严重。持续7 d治疗后,治疗组小鼠肠道双歧杆菌和乳酸杆菌数量明显上升,肠内容物挥发性脂肪酸含量明显上升,损伤的肠黏膜基本修复。结论应用林可霉素可以成功建立肠道微生态失调动物模型;马齿苋多糖具有扶植肠道正常菌群生长,调整菌群失调,防治感染的作用,是理想的中药微生态调节剂。     

Treatment and mechanism of fecal microbiota transplantation in mice with experimentally induced ulcerative colitis

Restoring intestinal microbiota dysbiosis with fecal microbiota transplantation is considered as a promising treatment for ulcerative colitis. However, the mechanisms underlying its relieving effects remain unclear. Ulcerative colitis pathogenesis is associated with the involvement of immune cells and inflammatory cytokines. Here, we aimed to investigate the effect of fecal microbiota transplantation on T cell cytokines in a dextran sulfate sodium-induced ulcerative colitis mouse model. Five-aminosalicylic acid (5-ASA) was used as the positive control. Male C57BL/6 mice were randomly assigned to control, model (UC), UC + FMT, and UC + 5-ASA groups. Each group consisted of five mice. The establishment of the mouse model was verified by fecal occult-blood screening and hematoxylin–eosin staining. Results showed that fecal microbiota transplantation reduced colonic inflammation, significantly decreased T helper (Th)1 and Th17 cells, interferon-gamma, interleukin-2 and interleukin-17, as well as significantly increased Th2 and regulatory T (Treg) cells, interleukin-4, interleukin-10, and transforming growth factor-beta, and improved routine blood count. Furthermore, 16S rRNA gene-sequencing analysis showed a significant increase in the relative abundance of genus Akkermansia and a significant decrease in the relative abundance of genus Helicobacter in the ulcerative colitis group. Fecal microbiota transplantation restored the profile of the intestinal microbiota to that of the control group. These findings demonstrated the capability of fecal microbiota transplantation in controlling experimentally induced ulcerative colitis by improving Th1/Th2 and Th17/Treg imbalance through the regulation of intestinal microbiota.     

The microbiota protects against Pseudomonas aeruginosa pneumonia via γδ T cell-neutrophil axis in mice

Exposure to antimicrobials leading to microbiota dysbiosis has been found to be an independent risk factor for extensively drug-resistant Pseudomonas aeruginosa acquisition. Microbiota dysbiosis may induce imbalanced immune responses and can affect disease susceptibility. However, the potential role of commensal microbiota in bacterial pneumonia is poorly defined. The aim of this study was to investigate the mechanistic basis for the defective host defenses against P. aeruginosa pneumonia induced by antibiotic pretreatment perturbing microbiota. We found that antibiotic pretreatment significantly perturbed the composition of intestinal microbiota. The microbiota dysbiosis impaired host defenses against P. aeruginosa pneumonia, as reflected by the increased bacterial burden and dissemination, compromised local inflammatory responses and shortened survival time in microbiota-depleted mice compared with controls. This impairment correlated with a defective γδ T17 cell and downstream neutrophil responses. Anti-TCRγδ-treated mice had changes similar to those in the microbiota-depleted mice. Overall, our results suggest the importance of microbiota in supporting the host defense against pneumonia, define a crucial role for the γδ T cell-neutrophil axis in the potential mechanism, and delineate the deleterious effects of antibiotic treatment on antibacterial defenses.     

High‐fat diet‐induced dysbiosis mediates MCP‐1/CCR2 axis‐dependent M2 macrophage polarization and promotes intestinal adenoma‐adenocarcinoma sequence

High‐fat diet (HFD) is a well‐known risk factor for gut microbiota dysbiosis and colorectal cancer (CRC). However, evidence relating HFD, gut microbiota and carcinogenesis is limited. Our study aimed to demonstrate that HFD‐induced gut dysbiosis promoted intestinal adenoma‐adenocarcinoma sequence. In clinical study, we found that HFD increased the incidence of advanced colorectal neoplasia (AN). The expression of monocyte chemoattractant protein 1 (MCP‐1), CC chemokine receptor 2 (CCR2) and CD163 in CRC patients with HFD was significantly higher than that in CRC patients with normal diet. When it comes to the Apc^min/+ mice, HFD consumption could induce gut dysbiosis and promote intestinal carcinogenesis, accompanying with activation of MCP‐1/CCR2 axis that recruited and polarized M2 tumour‐associated macrophages. Interestingly, transfer of faecal microbiota from HFD‐fed mice to another batch of Apc^min/+ mice in the absence of HFD could also enhance carcinogenesis without significant body weight gain and induced MCP‐1/CCR2 axis activation. HFD‐induced dysbiosis could also be transmitted. Meanwhile, antibiotics cocktail treatment was sufficient to inhibit HFD‐induced carcinogenesis, indicating the vital role of dysbiosis in cancer development. Conclusively, these data indicated that HFD‐induced dysbiosis accelerated intestinal adenoma‐adenocarcinoma sequence through activation of MCP‐1/CCR2 axis, which would provide new insight into better understanding of the mechanisms and prevention for HFD‐related CRC.     

Targeted Restoration of the Intestinal Microbiota with a Simple,Defined Bacteriotherapy Resolves Relapsing Clostridium difficile Disease in Mice

Relapsing C. difficile disease in humans is linked to a pathological imbalance within the intestinal microbiota, termed dysbiosis, which remains poorly understood. We show that mice infected with epidemic C. difficile (genotype 027/BI) develop highly contagious, chronic intestinal disease and persistent dysbiosis characterized by a distinct, simplified microbiota containing opportunistic pathogens and altered metabolite production. Chronic C. difficile 027/BI infection was refractory to vancomycin treatment leading to relapsing disease. In contrast, treatment of C. difficile 027/BI infected mice with feces from healthy mice rapidly restored a diverse, healthy microbiota and resolved C. difficile disease and contagiousness. We used this model to identify a simple mixture of six phylogenetically diverse intestinal bacteria, including novel species, which can re-establish a health-associated microbiota and clear C. difficile 027/BI infection from mice. Thus, targeting a dysbiotic microbiota with a defined mixture of phylogenetically diverse bacteria can trigger major shifts in the microbial community structure that displaces C. difficile and, as a result, resolves disease and contagiousness. Further, we demonstrate a rational approach to harness the therapeutic potential of health-associated microbial communities to treat C. difficile disease and potentially other forms of intestinal dysbiosis.     

Circadian Disorganization Alters Intestinal Microbiota

Intestinal dysbiosis and circadian rhythm disruption are associated with similar diseases including obesity, metabolic syndrome, and inflammatory bowel disease. Despite the overlap, the potential relationship between circadian disorganization and dysbiosis is unknown; thus, in the present study, a model of chronic circadian disruption was used to determine the impact on the intestinal microbiome. Male C57BL/6J mice underwent once weekly phase reversals of the light:dark cycle (i.e., circadian rhythm disrupted mice) to determine the impact of circadian rhythm disruption on the intestinal microbiome and were fed either standard chow or a high-fat, high-sugar diet to determine how diet influences circadian disruption-induced effects on the microbiome. Weekly phase reversals of the light:dark (LD) cycle did not alter the microbiome in mice fed standard chow; however, mice fed a high-fat, high-sugar diet in conjunction with phase shifts in the light:dark cycle had significantly altered microbiota. While it is yet to be established if some of the adverse effects associated with circadian disorganization in humans (e.g., shift workers, travelers moving across time zones, and in individuals with social jet lag) are mediated by dysbiosis, the current study demonstrates that circadian disorganization can impact the intestinal microbiota which may have implications for inflammatory diseases.