高脂饮食中添加短链菊粉对小鼠肠道菌群的影响

目的:探究高脂饮食中添加短链菊粉对小鼠肠道菌群的影响。方法:选择8周龄雄性小鼠,5只喂食高脂饲料,5只喂食10%菊粉复合型高脂饲料,喂食8周后收集小鼠粪便,检测小鼠粪便中三种主要的短链脂肪酸。同时,提取小鼠粪便中的细菌基因组,对菌群基因组16S rRNA基因V4高变区进行测序,对数据进行PCoA分析、Alpha多样性分析、LEfSe分析和16S功能预测。结果:菊粉添加后,小鼠粪便中含有的细菌DNA量增多,短链脂肪酸增加。菊粉组和对照组PCoA图可以看到明显聚类。菊粉组物种多样性低于对照组。菊粉组小鼠粪便中S24_7菌科丰度上升;Lachnospiraceae(毛螺菌科),Ruminococcaceae(瘤胃菌科)和Deferribacteraceae(脱铁杆菌科)丰度下降。16S基因功能预测发现22个第二层级的KEGG通路发生变化。结论:高脂饮食情况下短链菊粉的添加会改变小鼠肠道菌群,继而影响肠道菌群的功能。     

抗性淀粉对HFA小鼠肠道菌群的影响

目的 以人源菌群(HFA)小鼠为研究模型,观察抗性淀粉(RS)对高脂饮食诱导的肥胖小鼠肠道菌群的多样性的影响.方法 将30只无菌小鼠接种健康人志愿者的粪便悬液构建HFA小鼠模型后,随机分成3组,一组喂养含20％的抗性淀粉的高脂饲料(RS组),一组喂养纯高脂饲料(CK组),一组喂养普通饲料(CONV组),取第0周和第8周的小鼠新鲜粪便,用PCR-DGGE分析3组小鼠的肠道菌群的相似性和多样性.结果 3组小鼠在第0周时肠道菌群多样性的相似度达到79％～87％,与人的肠道菌群相似性达到39％,说明构建HFA小鼠模型成功,第8周时,3组之间的均匀度(E)和Shannon指数差异无统计学意义(P＞0.05),而丰富度(S)在高脂组(CK)与普通饲料组(CONV)和抗性淀粉组(RS)之间差异都有统计学意义(P＜0.05),说明高脂饮食引起肠道菌群多样性增加,而抗性淀粉则能降低这种多样性.结论 抗性淀粉可以显著影响HFA小鼠的肠道菌群多样性.     

茶籽皂苷对高脂血症大鼠肠道菌群的影响研究

本实验旨在研究茶籽皂苷对高脂血症大鼠肠道菌群的影响。60只雄性Wistar大鼠适应性喂养1周后随机分为正常组(10只,C组)和造模组(50只)。C组饲喂基础饲料,造模组饲喂高脂饲料。造模成功后,将造模组大鼠按体重随机分为模型对照组(H组)、阳性对照组(P组)、茶籽皂苷高、中、低剂量组(TSH组、TSM组、TSL组),并进行灌胃。灌胃4周后,用16S rRNA测序技术检测茶籽皂苷对肠道菌群的影响。结果显示:与C组相比,H组大鼠肠道菌群的丰度和多样性显著降低,其群落结构从门到属均发生显著变化。灌胃茶籽皂苷后,肠道菌群的丰度以及从门到属的群落结构也均得到有效调节,有益菌群相对丰度提升,致病菌群得到抑制。结果表明茶籽皂苷对由高脂饲料诱发的高脂血症大鼠的肠道菌群起到正向调节作用。     

西芹提取液对高脂小鼠肠道菌群平衡的影响

目的应用PCR-DGGE技术评价西芹对昆明小鼠肠道菌群平衡的影响。方法给予小鼠高脂饲料,复制高脂模型,模型复制成功后,每日给予小鼠西芹提取液灌胃,连续28 d。分别在实验的0、14和42 d,收集每只小鼠粪便,提取基因组DNA,应用PCR-DGGE(聚合酶链式反应——变性梯度凝胶电泳)技术获得肠道菌群分子指纹图谱,针对相似性、多样性分析及优势条带的序列进行分析。结果高脂饲料喂养后肠道益生菌约氏乳杆菌等趋近消失,出现了特异菌真杆菌,而西芹灌胃组和食用基础饲料组真杆菌数量显著减少,且西芹灌胃组作用大于食用基础饲料自然恢复组。结论西芹对高脂小鼠肠道菌群的平衡有显著的恢复作用。     

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。     

益生菌干预对高脂高糖饮食诱导肥胖小鼠肠道菌群及脂代谢影响的研究

目的探讨益生菌干预对高脂高糖饮食诱导肥胖小鼠肠道菌群及脂代谢的影响。方法 C57BL/6J雌性小鼠30只随机分为正常对照组、肥胖组和益生菌干预组,每组10只,分别给予标准饲料、高脂高糖饲料以及高脂高糖饲料同时给予益生菌干预,连续喂养6周,测量并分析三组小鼠的体重。留取小鼠粪便样本,应用PCR-DGGE法分析菌群,应用酶反应比色法分析三组小鼠血脂情况。结果与正常对照组小鼠相比,肥胖小鼠体重明显增加,益生菌干预组小鼠体重略有增加;肥胖组小鼠肠道菌群紊乱,与正常对照组分别聚为两大类,益生菌干预组小鼠肠道菌群与正常对照组聚为一大类。肥胖小鼠血清总胆固醇、低密度脂蛋白含量升高,益生菌干预组小鼠较肥胖组血清总胆固醇、低密度脂蛋白含量降低,但与正常对照组仍有差异。结论高脂高糖饮食诱导肥胖小鼠存在肠道菌群结构失调及脂代谢异常,益生菌干预可以改善肥胖小鼠菌群失调以及脂代谢紊乱。     

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。     

喂养方式对西安地区0～2个月婴儿肠道菌群的影响

目的 探讨母乳和混合喂养方式对西安地区婴儿肠道菌群的影响。方法 收集24例0～2月龄西安地区健康婴儿粪便样本,根据婴儿的喂养方式,将样本分为母乳喂养组（17例）和以奶粉为主的混合喂养组（7例）。利用16S rRNA基因测序技术对不同喂养方式的婴儿肠道菌群进行测序,比较不同喂养方式对婴儿肠道菌群多样性和菌群差异的影响。结果 母乳喂养组婴儿粪便样本Ace指数和Chao1指数显著高于混合喂养组（t=4.886, P<0.05; t=6.855, P<0.05）, Shannon指数显著低于混合喂养组（t=2.126,P<0.05）。门水平上,2组样本均以放线菌门、厚壁菌门、变形菌门和拟杆菌门为主,但比例存在差异。相比混合喂养组,母乳喂养组婴儿粪便样本放线菌门和拟杆菌门丰度显著升高（U=6,P<0.05;U=0,P<0.05）,厚壁菌门和变形菌门丰度显著降低（U=24,P<0.05;U=16,P<0.05）。属水平上,母乳喂养组婴儿粪便样本双歧杆菌相对丰度（72.04%）显著升高,同时发现罗氏菌属和葡萄球菌属相对丰度也高于混合喂养组（U=17,P<...     

口服肝素与小鼠肠道菌群的相互作用

口服肝素药物的开发需要系统地理解口服肝素与肠道菌群之间的互作过程。通过荧光体视镜观察荧光素标记的肝素经小鼠口服后在体内的分布情况,利用高效液相色谱法检测肝素在模拟胃肠液中的稳定性和体外培养肠道菌群模拟肠道菌对肝素的降解作用,发现口服肝素主要分布在小鼠胃肠道内,在体外模拟胃肠液条件下肝素结构稳定,但能够被添加肝素的厌氧培养基培养后的肠道菌群降解。为了进一步揭示口服肝素对健康小鼠肠道菌群的影响,利用Illumina MiSeq高通量测序技术测定口服肝素后C57BL/6J小鼠粪便菌群的16S rRNA序列,与口服生理盐水的小鼠粪便菌群进行对比,发现口服肝素的小鼠粪便菌群的生物多样性降低;在门水平上,菌群结构差异不显著;而在属水平上,别样杆菌属Alistipes、副萨特氏菌属Parasutterella和艾克曼菌属Akkermansia相对丰度增高,而嗜胆菌属Bilophila、肠杆菌属Enterorhabdus、瘤胃梭菌属Ruminiclostridium、普雷沃氏菌科Prevotellaceae_UCG_001、瘤胃梭菌属Ruminiclostridium-9、拟杆菌属Bacteroides、Lachnoclostridium、Candidatus_Saccharimonas、Intestinimonas和Dubosiella的相对丰度减少,表明口服肝素能够影响小鼠肠道菌群结构。此外,实验发现口服肝素对小鼠无明显毒副作用,具有较高安全性。研究结果将为开发肝素口服递送策略提供新的思路,为口服肝素类药物的开发提供参考。     

强化玉米饮食对小鼠肠道菌群结构和功能的影响

【目的】采用高通量测序方法研究强化玉米饮食对小鼠肠道菌群结构的影响以及可提高宿主糖代谢相关菌群功能基因的分析。【方法】分别给予两组小鼠(各10只)常规饮食和强化玉米饮食(1/4的玉米粉加3/4的常规饮食成分),喂养10周,之后采集小鼠粪便样本,提取DNA,使用高通量测序仪进行宏基因组测序分析,比较两组小鼠肠道菌群和功能基因的差异。【结果】两组小鼠的终末体重没有明显差异。各样本DNA的测序有效率足够,肠道菌群的多样性存在一定差异。属放线菌门(Actinobacteria)的双歧杆菌(Bifidobacteriales)-B.pseudolongum分支和Coriobacteriia-Collinsella/Enterorhabdus分支的丰度在强化玉米饮食组的小鼠中显著升高,相应的宏基因组中涉及糖酵解和胆汁酸合成的一些酶和功能单元的含量也在强化玉米饮食组显著升高。【结论】强化玉米饮食可以提高肠道菌群中双歧杆菌等益生菌的丰度,增加宏基因组糖脂代谢相关基因和通路的含量,从而可能促进宿主的糖代谢功能。     

人工辅助投食对滇金丝猴肠道微生物群落的影响

【背景】肠道菌群与宿主的消化吸收、免疫抵抗和行为等息息相关,并受宿主的饮食、生活环境等因素影响。【目的】人工辅助投食能增加野生动物的营养摄入,但对其肠道菌群影响的研究较少。【方法】以云南白马雪山国家级自然保护区内的野生和人工辅助投食滇金丝猴群的新鲜粪便为材料,通过高通量测序探究人工辅助投食对猴群肠道菌群的影响。【结果】人工辅助投食的猴群肠道菌群丰富度、均匀度及谱系多样性更高,并且个体间群落组成差异更小。通过多级物种差异判别分析(linear discriminant analysis effect size, LEfSe)分析发现,人工辅助投食对20种不同分类水平的细菌相对丰度有影响,包括提升了厚壁菌门(Firmicutes)等8种类群的相对丰度,降低了变形菌门(Proteobacteria)、拟杆菌门(Bacteroidetes)等12种类群的相对丰度。通过构建微生物相关网络发现,野生猴群肠道菌群网络结构更加复杂,鲁棒性更高。京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes, KEGG)功能预测结果表明,人工辅助投食降低了猴群肠...     

基于16S rDNA扩增子测序技术揭示真胃左方变位对奶牛粪便微生物的影响

[目的]本试验旨在测定产后健康奶牛和罹患真胃左方变位(left displacement of the abomasum, LDA)奶牛粪便中微生物菌群的变化,以期探讨LDA发生与菌群的关联性,评估其对机体代谢的潜在影响.[方法]采用16S rDNA高通量测序技术,测定10头健康奶牛(Health)、10头真胃左方变位...     

CD38 deficiency suppresses adipogenesis and lipogenesis in adipose tissues through activating Sirt1/PPARγ signaling pathway

It has been recently reported that CD38 was highly expressed in adipose tissues from obese people and CD38‐deficient mice were resistant to high‐fat diet (HFD)‐induced obesity. However, the role of CD38 in the regulation of adipogenesis and lipogenesis is unknown. In this study, to explore the roles of CD38 in adipogenesis and lipogenesis in vivo and in vitro, obesity models were generated with male CD38^?/? and WT mice fed with HFD. The adipocyte differentiations were induced with MEFs from WT and CD38^?/? mice, 3T3‐L1 and C3H10T1/2 cells in vitro. The lipid accumulations and the alternations of CD38 and the genes involved in adipogenesis and lipogenesis were determined with the adipose tissues from the HFD‐fed mice or the MEFs, 3T3‐L1 and C3H10T1/2 cells during induction of adipocyte differentiation. The results showed that CD38^?/? male mice were significantly resistant to HFD‐induced obesity. CD38 expressions in adipocytes were significantly increased in WT mice fed with HFD, and the similar results were obtained from WT MEFs, 3T3‐L1 and C3H10T1/2 during induction of adipocyte differentiation. The expressions of PPARγ, AP2 and C/EBPα were markedly attenuated in adipocytes from HFD‐fed CD38^?/? mice and CD38^?/? MEFs at late stage of adipocyte differentiation. Moreover, the expressions of SREBP1 and FASN were also significantly decreased in CD38^?/? MEFs. Finally, the CD38 deficiency‐mediated activations of Sirt1 signalling were up‐regulated or down‐regulated by resveratrol and nicotinamide, respectively. These results suggest that CD38 deficiency impairs adipogenesis and lipogenesis through activating Sirt1/PPARγ‐FASN signalling pathway during the development of obesity.     

Behavioural effects of high fat diet in a mutant mouse model for the schizophrenia risk gene neuregulin 1

Schizophrenia patients are often obese or overweight and poor dietary choices appear to be a factor in this phenomenon. Poor diet has been found to have complex consequences for the mental state of patients. Thus, this study investigated whether an unhealthy diet [i.e. high fat diet (HFD)] impacts on the behaviour of a genetic mouse model for the schizophrenia risk gene neuregulin 1 (i.e. transmembrane domain Nrg1 mutant mice: Nrg1 HET). Female Nrg1 HET and wild‐type‐like littermates (WT) were fed with either HFD or a control chow diet. The mice were tested for baseline (e.g. anxiety) and schizophrenia‐relevant behaviours after 7 weeks of diet exposure. HFD increased body weight and impaired glucose tolerance in all mice. Only Nrg1 females on HFD displayed a hyper‐locomotive phenotype as locomotion‐suppressive effects of HFD were only evident in WT mice. HFD also induced an anxiety‐like response and increased freezing in the context and the cued version of the fear conditioning task. Importantly, CHOW‐fed Nrg1 females displayed impaired social recognition memory, which was absent in HFD‐fed mutants. Sensorimotor gating deficits of Nrg1 females were not affected by diet. In summary, HFD had complex effects on the behavioural phenotype of test mice and attenuated particular cognitive deficits of Nrg1 mutant females. This topic requires further investigations thereby also considering other dietary factors of relevance for schizophrenia as well as interactive effects of diet with medication and sex.     

Deficiency in the anti‐aging gene Klotho promotes aortic valve fibrosis through AMPKα‐mediated activation of RUNX2

Fibrotic aortic valve disease (FAVD) is an important cause of aortic stenosis, yet currently there is no effective treatment for FAVD due to its unknown etiology. The purpose of this study was to investigate whether deficiency in the anti‐aging Klotho gene (KL) promotes high‐fat‐diet‐induced FAVD and to explore the underlying molecular mechanism. Heterozygous Klotho‐deficient (KL^+/?) mice and WT littermates were fed with a high‐fat diet (HFD) or normal diet for 13 weeks, followed by treatment with the AMPKα activator (AICAR) for an additional 2 weeks. A HFD caused a greater increase in collagen levels in the aortic valves of KL^+/? mice than of WT mice, indicating that Klotho deficiency promotes HFD‐induced aortic valve fibrosis (AVF). AMPKα activity (pAMPKα) was decreased, while protein expression of collagen I and RUNX2 was increased in the aortic valves of KL^+/? mice fed with a HFD. Treatment with AICAR markedly attenuated HFD‐induced AVF in KL^+/? mice. AICAR not only abolished the downregulation of pAMPKα but also eliminated the upregulation of collagen I and RUNX2 in the aortic valves of KL^+/? mice fed with HFD. In cultured porcine aortic valve interstitial cells, Klotho‐deficient serum plus cholesterol increased RUNX2 and collagen I protein expression, which were attenuated by activation of AMPKα by AICAR. Interestingly, silencing of RUNX2 abolished the stimulatory effect of Klotho deficiency on cholesterol‐induced upregulation of matrix proteins, including collagen I and osteocalcin. In conclusion, Klotho gene deficiency promotes HFD‐induced fibrosis in aortic valves, likely through the AMPKα–RUNX2 pathway.     

饲养方式对藏猪结肠消化酶活性、菌群结构及短链脂肪酸含量的影响

【目的】为探究饲养方式对藏猪结肠消化酶活性、菌群结构和短链脂肪酸含量的影响。【方法】研究分别选取5头相同月龄的放养藏猪和舍饲藏猪。屠宰采集结肠粪便样品,分别利用酶联免疫吸附法(enzyme linked immunosorbent assay, ELISA)试剂盒、高通量测序技术和气相色谱仪测定放养藏猪和舍饲藏猪结肠消化酶活性、菌群结构和短链脂肪酸含量。【结果】同一月龄下,放养藏猪的日增重显著低于舍饲藏猪(P<0.05)。放养藏猪结肠中纤维素酶和半纤维素酶的活性均显著高于舍饲藏猪(P<0.05);2种饲养方式藏猪结肠的6种α多样性指数均无显著差异(P>0.05),且主成分分析(principal component analysis, PCA)得到放养藏猪和舍饲藏猪结肠菌群存在一定的相似性。在门和科分类水平上,相较于舍饲藏猪,放养藏猪结肠中疣微菌门、黄杆菌科、月形单胞菌科、浮霉状菌科和伊格尔兹氏菌科的相对丰度显著升高,而链球菌科、韦荣氏球菌科、假单胞菌科、红环菌科、红螺菌科、乳杆菌科、理研菌科和巴斯德氏菌科的相对丰度显著降低(P<0.05);在属和种分类水平上,...     

Mice lacking myotubularin-related protein 14 show accelerated high-fat diet-induced lipid accumulation and inflammation

The phosphoinositide phosphatase, myotubularin-related protein 14 (MTMR14), has been reported to play an important role in the regulation of muscle performance, autophagy, and aging in mice. We previously showed that MTMR14-knockout (KO) mice gain weight earlier than their wild-type (WT) littermates even on a normal chow diet (NCD), suggesting that this gene might also be involved in regulating metabolism. In the present study, we evaluated the effect of MTMR14 deficiency on high-fat diet (HFD)-induced obesity, lipid accumulation, metabolic disorders, and inflammation in WT and MTMR14-KO mice fed with NCD or HFD. To this end, MTMR14-KO mice fed with HFD showed significantly increased body weight, blood glucose levels, serum triglyceride (TG) levels, and total cholesterol (TC) levels as compared to their age-matched WT control. Additionally, lipid accumulation also increased in the KO mice. Simultaneously, the expression of metabolism-associated genes (Glut4, adiponectin, and leptin) was different in the liver, muscle, and fatty tissue of MTMR14-KO mice fed with HFD. More importantly, the expression of several inflammation-associated genes (TNF-α, IL-6, IL-1β, and MCP-1) dramatically increased in the liver, muscle, and fatty tissue of MTMR14-KO mice relative to control. Taken together, these results suggest that MTMR14 deficiency accelerates HFD-induced metabolic dysfunction and inflammation. Furthermore, the results showed that exacerbated metabolic dysfunction and inflammation may be regulated via the PI3K/Akt and ERK signaling pathways.     

探讨不同浓度的清郁和降汤对反流性食管炎大鼠肠道菌群的影响

【目的】观察不同浓度的清郁和降汤对反流性食管炎（reflux esophagitis，RE）的治疗效果，并探讨其对肠道菌群的影响。【方法】将36只健康雄性SD大鼠随机分为6组，其中1组为假手术组，剩余5组大鼠采用“前胃结扎+外置幽门部分结扎术”手术造模方法建立反流性食管炎模型。造模2周后将术后全部存活的30只大鼠随机分成对照组、中药高剂量组（予高剂量清郁和降汤）、中药中剂量组（予中剂量清郁和降汤）、中药低剂量组（予低剂量清郁和降汤）、西药组（予泮托拉唑钠肠溶胶囊+枸橼酸莫沙比利分散片+复方嗜酸乳杆菌片），每组6只。于术后第15天开始灌胃，其中假手术组及对照组予蒸馏水灌胃，其他组分别给予相应的药物灌胃，持续灌胃14 d后将所有大鼠处死后进行取材。以苏木精-伊红（hematoxylin-eosin，HE）染色观察大鼠食管组织的病理学改变；采用16S rRNA基因高通量测序检测其肠道黏膜的菌群构成。【结果】反流性食管炎大鼠存在着较为明显的肠道菌群结构变化及肠道菌群多样性较低的情况，B组（对照组）中厚壁菌门和拟杆菌门占比减少，变形菌门占比增多，且假单胞菌属、青枯菌属等细菌增多。低、中、高3种浓度的清郁和降汤均能够提升RE大鼠的肠道菌群多样性，增加拟杆菌门及厚壁菌门，降低变形菌门的占比，从属水平上看，清郁和降汤能够提升大鼠肠道中拟杆菌属、乳杆菌属、瘤胃球菌属、颤螺旋菌属、双歧杆菌属和狄氏副拟杆菌属等益生菌占比。以D组（中剂量组）对大鼠肠道菌群多样性提升最为明显，其效果最接近假手术组。特征微生物方面，B组以变形菌门为特征性微生物，D、E两组出现了放线菌门及拟杆菌门下属细菌为特征微生物的情况，在门水平与F组相同。【结论】清郁和降汤能够有效地治疗反流性食管炎，其机制可能与改变RE大鼠的肠道菌群结构、减少有害菌、提升益生菌的占比和改善肠道菌群多样性有关。