合生元益生菌冲剂对小鼠肠道菌群调节作用的研究

目的 研究合生无益生菌冲剂对小鼠肠道菌群的调节作用.方法 将小鼠分为阴性对照组、合生元益生菌冲剂组.阴性对照组灌服蒸馏水14 d,合生元益生菌冲剂组灌服1 g/kg·d剂量的台生元益生菌冲剂14 d,检测实验前后肠道菌群数量.结果 灌服后小鼠肠道菌群与灌服前比较,,双歧杆菌数量有显著性增加（P＜0.05）,肠杆菌、肠球菌、乳杆菌和产气荚膜梭菌数量有无显著性变化（P＞0.05）.结论 合生元益生菌冲剂对小鼠肠道菌群具有一定的调节作用.     

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。     

玉米紫色植株花色苷色素对小鼠肠道菌群的影响

目的了解玉米紫色植株花色苷色素对小鼠肠道菌群的影响。方法按中华人民共和国卫生部《保健食品检验与评价技术规范》中的动物实验调节肠道菌群功能的检验方法,观察玉米紫色植株色素对小鼠肠道中肠杆菌、乳杆菌、双歧杆菌数量的影响。结果灌服后小鼠肠道乳杆菌、双歧杆菌数量变化差异均无显著性,低剂量组肠扦菌数量明显低于灌服前（P〈0．05）,其他组肠杆菌数量变化差异无显著性。结论玉米紫色植株色素对小鼠肠道菌群可能有影响。     

中药人参合剂对提高小鼠定植抗力作用的初步研究

本实验用肠炎杆菌攻击正常小白鼠,肠炎杆菌菌液浓度为１０８个／ｍｌ,灌喂０．２ｍｌ,造成肠炎杆菌在肠道内大量定植后,分别用中药人参合剂及复方新诺明（ＣＯ－ＳＭＺ）治疗,连续６天后,观察对比两种药物对受攻击小鼠的治病效果。检测小鼠粪便中的四种菌群成员,肠炎杆菌、肠杆菌、双歧杆菌和乳杆菌。结果表明：中药人参合剂治疗组肠炎杆菌显著下降,与ＣＯ－ＳＭＺ组相似（Ｐ＞０．０５）,但双歧杆菌与乳杆菌显著升高,与复方新诺明组相比有显著性差异（Ｐ＜０．０５）。我们认为中药人参合剂治疗肠道感染的主要机制是中药人参合剂扶植正常菌群生长,提高小鼠的定植抗力的作用。     

三联益生菌对便秘小鼠肠道推进功能改善及其对 小鼠肠道菌群调节作用的研究

目的研究新型嗜酸乳杆菌NCFM-乳双歧杆菌Bi-07-鼠李糖乳杆菌NH001三联益生菌制剂对便秘小鼠小肠蠕动的促进作用,以及其对小鼠肠道菌群构成比例的调节作用。方法小肠推进率实验:将50只KM小鼠适应性饲养后随机分为空白对照组、模型对照组和低中高三个剂量组。空白对照组和模型对照组给予蒸馏水,低中高剂量组每天分别给予0.165、0.330、0.990g/(kg·bw)益生菌制剂,灌胃14d后使用复方地芬诺酯建立小鼠便秘模型观察小肠墨汁推进率;肠道菌群调节作用实验:将42只BALB/c小鼠中随机抽取10只作为自身对比空白组,适应性饲养后将所有小鼠随机分为空白对照组和低中高三个剂量组,空白对照组给予蒸馏水,各剂量组以相同剂量给予益生菌制剂灌胃。自身对比空白组于灌胃前无菌采集小鼠直肠粪便,所有小鼠灌胃30d后无菌采集直肠粪便,使用16SrDNA基因测序对粪便中菌群DNA进行多样性及各水平菌群物种测定。结果高剂量组便秘小鼠小肠推进率显著高于模型对照组(P0.05);各剂量组肠道菌群多样性、双歧杆菌属、乳杆菌属数量较灌胃前显著增加(P0.05),大肠埃希菌属、梭菌属、肠球菌属较灌胃前显著降低(P0.05)。结论三联益生菌能够有效促进便秘小鼠肠道蠕动能力,改善便秘症状;同时对小鼠肠道菌群结构有调节作用。     

罗伊乳杆菌对小鼠血清细胞因子含量和 肠道菌群的影响

目的探讨罗伊乳杆菌对小鼠免疫力及肠道菌群的影响。方法将50只昆明系小鼠按体重随机分成5组:空白对照组、低剂量组、中剂量组、高剂量组和发酵上清液组。空白对照组灌胃等量生理盐水,其余各组分别灌胃罗伊乳杆菌菌液及发酵液上清21d后,眼球取血收集血清,采用ELISA法检测IL-2、IFN-γ、IgA和IgG含量;采用16SrRNA基因高通量测序分析小鼠肠道菌群变化。结果与对照组相比,其余各组小鼠血清IL-2、IFN-γ、IgA和IgG的水平均明显升高,粪便菌群丰度有所增高,多样性降低。与对照组相比,低剂量组和中剂量组小鼠粪便菌群结构无显著差异,而高剂量组和发酵上清液组小鼠粪便菌群结构差异显著。结论罗伊乳杆菌可以增强小鼠免疫力,提高菌群丰度,改变小鼠肠道菌群结构。     

螺旋藻对腹泻模型小鼠肠道菌群的影响

为评价螺旋藻对肠道菌群的影响, 采用注射氨苄青霉素的方法, 制造小鼠腹泻模型, 造模成功后灌服不同剂量螺旋藻进行治疗, 取不同治疗时间的粪便样品, 检测双歧杆菌、乳杆菌、大肠杆菌、肠球菌、拟杆菌、产气荚膜梭菌菌群的数量,并在螺旋藻灌胃8 d后, 测定小鼠不同肠段各指标菌群的数量。结果表明, 中高剂量组螺旋藻对腹泻模型小鼠肠道菌群有明显调整作用, 有效缩短了肠道菌群由失调到平衡的时间;粪便样品同各肠段内容物所反映的菌群的变化趋势相同, 即治疗恢复组的双歧杆菌、乳杆菌数量总体上高于生理盐水组, 肠杆菌、肠球菌、拟杆菌、产气荚膜梭菌数量均低于生理盐水组, 但取样位置不同, 各指标菌群的数量明显存在差异。     

地衣芽胞杆菌BL63516对抗生素诱导的 菌群紊乱小鼠肠黏膜屏障功能的影响

目的探索地衣芽胞杆菌BL63516对肠道菌群和肠黏膜屏障功能调节的效果和可能机制,为临床疾病的治疗提供新思路。方法采用健康清洁级雄性BALB/C小鼠30只构建动物模型,将小鼠分为3组(每组10只),分别为对照组(CON组)、头孢曲松钠灌胃组(CS组)和地衣芽胞杆菌BL63516治疗组(BL组),除CON组外其余两组小鼠给予300 mg/mL头孢曲松钠0.2 mL,2次/d灌服8 d,短期大量服用抗生素以建立肠道菌群紊乱模型。第9天起BL组小鼠给予地衣芽胞杆菌干预连续8 d。第17天摘眼球处死小鼠,取血、结肠、粪便样本。ELISA方法检测小鼠血清及肝脏组织中的细胞因子及LPS水平。PCR-变性梯度凝胶电泳方法及16S rRNA高通量测序测定小鼠肠道菌群结构。结果地衣芽胞杆菌干预后小鼠血清LPS水平较模型组显著降低,屏障功能改善。且地衣芽胞杆菌治疗后BL组小鼠肠道菌群丰富度和多样性显著升高,菌群结构有所改变。结论实验结果提示地衣芽胞杆菌BL63516可以有效改善抗生素所致的肠道菌群失调,对肠屏障功能有一定调节作用。     

富硒猴头菇对小鼠肠道菌群的调节作用

目的以昆明小鼠为实验对象,观察富硒猴头菇对小鼠肠道菌群的影响。方法设高、中和低3个剂量组和1个普通猴头菇对照组,1个蒸馏水对照组,搜集粪便,用细菌16S rDNA荧光定量PCR法测定3大类肠道菌的数量。结果富硒猴头菇冻干粉灌胃后,中剂量组和高剂量组小鼠肠道中大肠埃希菌明显减少,低、中剂量组双歧杆菌增多,高剂量组乳酸菌增多,差异有统计学意义(P0.05)。且断药3 d后,中剂量组的大肠埃希菌、双歧杆菌数量,高剂量组的大肠埃希菌、乳酸菌数量与正常对照组相比差异仍有统计学意义。结论富硒猴头菇对小鼠肠道菌群有明显调节作用。     

长短链菊粉对抗生素诱导肠道菌群紊乱小鼠调节作用的研究

目的采用头孢曲松钠诱发小鼠肠道菌群紊乱建立模型,观察不同剂量长短链菊粉对抗生素诱导的肠道菌群紊乱小鼠的调节作用。方法 2g/(kg·d)头孢曲松钠灌胃小鼠,连续8d,诱导小鼠肠道菌群紊乱,然后将菌群紊乱小鼠分为低剂量短链菊粉处理组、高剂量短链菊粉处理组、低剂量长链菊粉处理组、高剂量长链菊粉处理组和自然恢复组,连续处理2周,采用PCR-DGGE和菌落计数的方法观察各组小鼠肠道菌群的恢复情况。结果抗生素处理后,小鼠肠道菌群与正常小鼠相比具有显著差异,菌群条带数明显减少(P0.05),有益菌肠杆菌、乳酸杆菌、双歧杆菌和消化球菌含量明显减少(P0.05),条件致病菌葡萄球菌和铜绿假单胞菌含量明显增多(P0.05);经菊粉处理后,小鼠的肠道菌群恢复效果明显好于自然恢复组(P0.05),其中长链菊粉处理组优于短链菊粉处理组,且高剂量长链菌粉处理组恢复程度优于低剂量长链处理组。结论菊粉能够有效调节抗生素引起的肠道菌群紊乱,而长链菊粉的调节作用优于短链菊粉,且高剂量更有效。     

小檗碱对肠易激综合征大鼠肠道菌群的影响

目的通过口服小檗碱(Berberine,BBR)治疗大鼠肠易激综合征(IBS),初步研究小檗碱对IBS大鼠肠道菌群的影响。方法用避水应激试验制作大鼠IBS模型,分别给予模型大鼠小剂量(25mg/kg)小檗碱、大剂量(100mg/kg)小檗碱和利福昔明干预治疗10d,检测大鼠肠道菌群丰度、生物多样性及菌群结构组成的变化。结果避水应激试验造模后模型组大鼠排便次数增多、腹外斜肌EMG明显升高,肠道可见微炎症表现。小檗碱干预后,IBS大鼠排便增多情况、内脏高敏感性和肠道微炎症情况均显著改善,其中大剂量小檗碱组优于小剂量组。经小檗碱干预后,大鼠肠道菌群多样性显著降低,大剂量小檗碱抑菌作用强于小剂量小檗碱。大剂量小檗碱可显著提高IBS大鼠肠道乳杆菌科细菌的比例,同时降低肠杆菌科细菌的比例。结论小檗碱可以显著降低IBS大鼠内脏高敏感性,改善肠道微炎症,减少IBS大鼠排便增多症状。小檗碱可以调节肠道菌群丰度和多样性,且剂量越大,抑菌作用越明显。肠道乳杆菌科细菌在小檗碱治疗肠易激综合征的过程中可能起一定作用。     

Impacts of functional oligosaccharide on intestinal immune modulation in immunosuppressive mice

In order to research the role of soybean oligosaccharides (SBOSs) on improvements in the microenvironment of intestinal flora and immune function of cyclophosphamide (CTX) immunosuppressive mice. Via giving intragastric administration of Soybean oligosaccharide (SBOS) at the low dose (50/(kg·BW)/d), the middle dose (200 mg/(kg·BW)/d) and the high dose (500 mg/(kg·BW)/d) partly once a day, which is also 28 days in a row. At the same time, (SBOS) mice in the drug group and (CG) mice in the positive control group were given intraabdominal injection of CTX (200 mg/kg/d).The immunosuppressive mouse model (CY) was established after 72 h in the model group and the positive control group (CG) was given intragastric administration of levamisole hydrochloric acid (LMS) for 3 days, with the data of 80ug/kg/d after injection of CTX (for actually 72 h). On the 8th, 15th and 22nd day, the number of Bifidobacterium, Lactobacillus, Enterococcus and Clostridium perfringens m in the feces of mice in each dose of drug group were determined. After the test resulted, the cellular immune function, humoral immune function, monocyte/macrophage function, NK cell activity and cytokine secretion (tumor necrosis factor-α, interferon-gamma and IL-4) were measured in immunosuppressive mice each group. The results showed that 200 mg/(kg BW) soybean oligosaccharide could significantly promote the proliferation and inhibit the increase of Enterococcus in immunosuppressive mice. The soybean oligosaccharide of 500 mg/(kg BW) could dramatically promote the proliferation of both Bifidobacillus and Lactobacillus, and also inhibit the increase of both Enterobacteriaceae and Enterococcus in immunosuppressive mice. The regulatory function of SBOS on intestinal flora was positive. Soybean oligosaccharide (500 mg/(kg BW) could significantly promote the proliferation of Bifidobacillus and Lactobacillus in immunosuppressive mice and inhibit the increase of Enterococcus and Enterococcus. The proliferation of spleen lymphocytes induced by ConA, LPS in immunosuppressive mice was dose-dependent. But it was still lower than that of the normal group (CG0) (p > 0.05). The serum hemolysin level of immunosuppressive mice was significantly increased in each dose group (p < 0.05), and the level of antibody forming cells in spleen cells of each dose group was significantly increased (P < 0.05), and the level of antibody forming cells in spleen cells of each dose group was significantly higher than that of low dose group (p < 0.005), and the level of serum hemolysin in immunosuppressive mice was significantly increased in each dose group (p < 0.05). In the detection of immune effector cell activity in immunosuppressive mice, the phagocytic function of macrophages in high dose group and the natural killing activity of spleen NK cells in high dose drug group were significantly increased, which were not significantly different from those in positive control group (P < 0.05), but the expression of TNF-α, INF-γ and IL-4 cytokines in serum was increased in a dose dependent manner (p < 0.05). In conclusion, soybean oligosaccharide can significantly increase the diversity of intestinal microecology, increase the number of intestinal beneficial bacteria, has a correlation with the proliferation of Bifidobacterium and Lactobacillus in the intestinal tract, and inhibit the proliferation of harmful bacteria. The results showed that SBOS had a direct effect on the proliferation of intestinal flora under immunosuppression. Based on the improvement of intestinal microenvironment in immunosuppressive mice by soybean oligosaccharide for 25 days, the results showed that compared with the positive control group, the nonspecific and specific immunity of immunosuppressive mice in the drug group had a regulatory effect, which improved the phagocytic function of monocytes/macrophages, developed the level of antibody forming cells, enhanced the standard of the killing activity of NK cells, and promoted the expression of cytokines as well. Compared with the model group, the transformation and proliferation of spleen lymphocytes in the high and middle dose groups were remarkably increased, but all of the indexes did not reach the level of the normal blank group. By studying the improvement of intestinal microenvironment in immunosuppressive mice, to some extent, it is concluded that the proliferation of intestinal flora can improve the immunomodulatory function of the body, but it still lowers the normal immune degree, which reflects the immunomodulatory effect of the body on the stimulation of continuous external intake. The results demonstrate that the immunomodulatory ability of immunosuppressive body was insensitive to SBOS and provided a theoretical basis for the study of health care function of intestinal microenvironment improvement when SBOS acted on abnormal immune function. The results also improved the practical application value of SBOS.     

Autochthonous intestinal bacterial flora and cholesterol levels in specific pathogen-free swine fed high-lipid and high-sucrose diets

Graber, C. D. (Baylor University College of Medicine, Houston, Tex.). R. W. Moore, M. Suzuki, W. E. Redmond, R. M. O'Neal, and B. M. Lockhart. Autochthonous intestinal bacterial flora and cholesterol levels in specific pathogen-free swine fed high-lipid and high-sucrose diets. J. Bacteriol. 92:1290-1297. 1966.-Thirty-two specific pathogen-free (SPF) and conventional swine fed high fat, high sugar, and a standard ration were cultured for intestinal flora, and their blood cholesterol levels were measured. The diets, whether sterilized or not sterilized, fed ad libitum or restricted, did not alter bacterial flora greatly or influence blood cholesterol levels. Anaerobes outnumbered aerobes by several logs. Four autochthonous bacteria, lactobacilli, Escherichia coli, enterococci, and gram-variable, nonspore-forming anaerobes (GVNSA; a type of bacteroides), were shown to be constantly present in all animals regardless of dietary conditions. From the duodenum and jejunum of 14 pigs, GVNSA and Bacteroides nigrescens were cultured in rather large numbers, a finding not previously reported in swine or in most other mammals. This finding may have special significance in reference to bile acid and cholesterol metabolism.     

鱼油对断奶大鼠脏器指数及肠道菌群影响的研究

目的探讨鱼油对断奶大鼠脏器指数及肠道菌群的影响。方法选取（21±3）d日龄断奶SD大鼠96只,雌雄各半,随机分为2组,实验组饲喂添加0．5％鱼油的饲料,对照组饲喂正常饲料。分别于第7、14、21、28天处死各组大鼠12只,分别测定大鼠肝脏指数、胸腺指数及脾脏指数;应用梯度稀释法和培养法测定大鼠4种肠道正常菌群,即肠杆菌、双歧杆菌、乳酸杆菌、葡萄球菌。结果实验组第7天脾脏指数与对照组相比差异有显著性（P〈0．05）,第28天肝脏指数和胸腺指数与对照组相比差异有显著性（P〈0．05）;实验组第7、14和21天肠道内肠杆菌数量与对照组相比差异有显著性（P〈0．05）,第7天大鼠肠道内葡萄球菌数量与对照组相比差异有显著性（P〈0．05）,第14天大鼠肠道内葡萄球菌数量与对照组相比差异有非常显著性（P〈0．01）,实验组大鼠肠道内乳酸杆菌和双歧杆菌数量与对照组相比虽有所增高,但差异无显著性（P〉0．05）。结论鱼油可以明显提高断奶大鼠的脏器指数,并且可通过增加断奶大鼠肠道内双歧杆菌和乳酸杆菌数量,降低肠道内肠杆菌和葡萄球菌数量,进而调节断奶大鼠的肠内菌群,改善肠道内环境。     

组方食品对2型糖尿病小鼠血糖及 肠道菌群的影响

目的研究含有浒苔等植物的功能性食品对糖尿病小鼠血糖浓度的影响,并应用PCR-DGGE技术评价其对昆明小鼠肠道菌群稳态的影响。方法采用高脂饲料喂养昆明小鼠加腹腔注射链脲佐菌素(STZ)的方法建立糖尿病小鼠模型;将实验动物随机分为正常对照组、自然恢复组和功能性食品喂养组,连续给药4周,尾静脉采血测量血糖水平。收集小鼠新鲜粪便,提取粪便细菌基因组DNA,通过PCR-DGGE获得细菌群落指纹图谱,并进行相关软件分析,同时切离差异显著条带进行序列分析。结果本实验采用的功能性食品对糖尿病小鼠有降糖作用,并使血糖值稳定地维持在较低水平。4周后,功能性食品喂养的糖尿病小鼠血糖水平相对普通饲料喂养的糖尿病小鼠血糖发生显著性下降(t=4.19,P0.01);给2型糖尿病小鼠提供普通饲料时,其肠道菌群种类和数目相对较少;而提供功能性食品时,肠道菌群种类和数目相对增多,特别是双歧杆菌、Prevotella oryzae、Barnesiella intestinihominis、Culturomica和Parabacteroides distasonis明显增多,但是Muribaculum较少。结论高脂饲料结合STZ诱导的2型糖尿病小鼠肠道菌群发生显著改变,组方食品通过扶持肠道菌群,降低血糖水平。     

绿茶摄入量对小鼠肠道微生物及酶活性的影响

目的从肠道微生态角度研究绿茶摄入量对小鼠肠道微生物和酶活性的影响,探析饮茶(绿茶)有益健康的机制,为饮茶做出科学指导。方法将SPF级实验小鼠随机分为正常对照组、低浓度绿茶摄入组、中浓度绿茶摄入组和高浓度绿茶摄入组。低、中、高浓度绿茶摄入组分别给予10%、20%和30%(g/g)的茶水溶液,连续灌胃1个月后采集空肠至回肠段内容物进行微生物和酶活性分析,同时观察实验动物体质量变化情况。结果各实验组小鼠体质量增长与对照组相比无明显差异;与对照组相比,低浓度绿茶摄入组小鼠肠道乳酸菌数降低而细菌数升高,中浓度摄入组乳酸菌数、细菌以及双歧杆菌数显著降低,高浓度摄入组细菌数、双歧杆菌数升高而乳酸菌数显降低著,且中浓度摄入组乳酸菌数和细菌数均极显著地低于低浓度摄入组;由酶活性分析可看出,与对照组小鼠相比,各实验组小鼠肠道淀粉酶和蛋白酶活性均升高而木聚糖酶活性显著降低。结论摄入一定量的绿茶对肠道微生物有一定的调节作用,可提高淀粉酶和蛋白酶活性而益于机体健康。     

甲壳低聚糖的微生态学效应

目的 :了解甲壳低聚糖的微生态学效应 ,为其开发为微生态调节剂提供实验依据。方法 :用甲壳低聚糖〔6 0 0 m g/ (kg· d)〕给正常小鼠、肠道脱污染模型小鼠及 STZ糖尿病模型小鼠连续灌胃 7d及 2 1d,用添加 0 .3%甲壳低聚糖的高脂饲料为高血脂症大鼠模型连续灌胃 4 2 d。各空白对照组均用等体积无菌盐水灌胃。而后分析各组动物肠菌群 ,并测定 STZ糖尿病小鼠血糖、高血脂症大鼠 TC及 HDL- C值。结果 :甲壳低聚糖可以优化正常小鼠肠菌群 ,双歧杆菌数较空白对照组明显增多 (P<0 .0 5 ) ,B/ E值为对照组19.3倍 ,可纠正实验动物因肠道脱污染、糖尿病及高血脂症所致的肠菌群失调 ,双歧杆菌或乳杆菌相应增加 ,与相应空白对照组比较 ,其差异均有显著性意义 ,可以明显降低糖尿病小鼠血糖及高血脂症大鼠血清TC值。结论 :甲壳低聚糖具有良好的微生态学效应 ,可作为微生态调节剂开发应用