Identification of a selective inhibitor of murine intestinal alkaline phosphatase (ML260) by concurrent ultra-high throughput screening against human and mouse isozymes

Alkaline phosphatase (AP) isozymes are present in a wide range of species from bacteria to man and are capable of dephosphorylation and transphosphorylation of a wide spectrum of substrates in vitro. In humans, four AP isozymes have been identified—one tissue-nonspecific (TNAP) and three tissue-specific—named according to the tissue of their predominant expression: intestinal (IAP), placental (PLAP) and germ cell (GCAP) APs. Modulation of activity of the different AP isozymes may have therapeutic implications in distinct diseases and cellular processes. For instance, changes in the level of IAP activity can affect gut mucosa tolerance to microbial invasion due to the ability of IAP to detoxify bacterial endotoxins, alter the absorption of fatty acids and affect ectopurinergic regulation of duodenal bicarbonate secretion. To identify isozyme selective modulators of the human and mouse IAPs, we developed a series of murine duodenal IAP (Akp3-encoded dIAP isozyme), human IAP (hIAP), PLAP, and TNAP assays. High throughput screening and subsequent SAR efforts generated a potent inhibitor of dIAP, ML260, with specificity for the Akp3-, compared to the Akp5- and Akp6-encoded mouse isozymes.     

乳杆菌微小膜蛋白对肠上皮细胞Caco-2的细胞毒性研究

目的 利用脂多糖（lipopolysaccharide,LPS）刺激模拟体外炎症环境,观察不同浓度下乳杆菌微小膜蛋白（micro integral membrane protein,MIMP）对肠上皮细胞Caco-2的生物学影响,评估其细胞毒性作用。方法 首先通过CCK-8实验检测在LPS刺激后不同浓度MIMP（0.01、0.1和1 ng/mL）对Caco-2细胞增殖活性的影响,并利用Toll样受体4（Toll-like receptor 4,TLR4）抑制剂作为阳性对照。其次利用流式细胞术检测不同浓度下MIMP对Caco-2细胞凋亡及细胞周期的影响。结果 在12 h的特定孵育时间内,单独应用不同浓度MIMP及TLR4抑制剂对Caco-2细胞增殖活性无显著影响（P＞0.05）,但是MIMP可以拮抗LPS对Caco-2细胞的促增殖作用（P<0.05）。不同浓度的MIMP对Caco-2细胞的周期和凋亡无明显影响（P＞0.05）。结论 不同浓度MIMP对Caco-2细胞的增殖、凋亡及细胞周期无明显影响,并可以拮抗LPS的促细胞增殖作用,因此具有较高的安全性,有望用于临床对炎症性肠病的治疗。     

Altered morphology in cultured rat intestinal epithelial IEC-6 cells is associated with alkaline phosphatase expression

Non-transformed, rat intestinal epithelial cells (IEC-6), and human intestinal colonic carcinoma cells (CACO-2) have both been used to study processes of epithelial cell differentiation. However, only CACO-2 cells have been described as spontaneously expressing phenotypic changes of differentiation in culture. We report here that when IEC-6 cells are grown in post-confluent culture, they develop structural changes similar to those seen in cells induced to differentiate by culture on Englebreth-Holm-Swarm (EHS) extracellular matrix proteins. Correlated with this morphological change is loss of nuclear localization of c-myc protein and development of cell surface alkaline phosphatase (ALP) enzymatic activity. Messenger RNAs for liver and intestinal isoforms of ALP were expressed in both pre- and post-confluent cells. Inhibition of ALP activity in post-confluent cells by levamisole indicated the expressed ALP activity to be of the liver isoform. We suggest the expression of ALP activity, which occurs concomitantly with morphological alterations in post-confluent IEC-6 cells, represents increased expression and localization to the cell surface of the liver isoform of ALP. Cultured IEC-6 cells may provide a non-transformed, in vitro alternative to CACO-2 cells for study of epithelial cell differentiation.     

溃疡性结肠炎患者肠道菌群分布的研究现状

溃疡性结肠炎(Ulcerative colitis,UC)的发病被认为是宿主遗传易感性、黏膜免疫与肠道菌群共同作用的结果。许多临床研究显示,与正常人相比,UC患者存在不同程度的菌群失调。艰难梭菌、致病性大肠埃希菌等致病微生物被怀疑与UC的发病相关,但目前还未找到充分证据证明它们与UC患者肠道炎症间的因果关系。就UC患者肠道菌群分布的研究现状、肠道菌群检测方法及未来研究进展进行了阐述。     

The unfolding intermediate state of calf intestinal alkaline phosphatase during denaturation in guanidine solutions

The equilibrium unfolding of calf intestinal alkaline phosphatase in guanidinium chloride (GdmCl) solutions was studied by following the fluorescence and ultraviolet difference spectra. At low concentrations of GdmCl (< 1.6 M), the fluorescence intensity decreased with a slight red shift of the emission maximum from 332 nm to 344 nm. An unfolding intermediate state was observed at a broad concentration range of GdmCl as a denaturant (between 1.6 and 2.6 M). This intermediate was characterized by increased fluorescence emission intensity, ultraviolet difference absorption at 236 nm and 260 nm, as well as increased binding to the protein and red shift of the fluorescence probe 1-anilinonaphthalene-8-sulfonic acid.     

地衣芽胞杆菌BL63516对DSS诱导的小鼠结肠炎的调节作用

目的 探索地衣芽胞杆菌BL63516对DSS诱导的小鼠结肠炎的缓解作用和可能机制,为临床疾病的治疗提供新思路。方法 用BALB/C雌性小鼠构建动物模型,将30只小鼠分为CON组、DSS组和BL组,每组10只,除CON组外,其余两组均给予3%（m/v）DSS自由饮用,并给予地衣芽胞杆菌BL63516干预。第8天摘眼球处死小鼠,取血、结肠、粪便样本。以ELISA法检测小鼠血清和结肠研磨液中的细胞因子。用PCR-变性梯度凝胶电泳（PCR-DGGE）方法分析小鼠肠道菌群结构。结果 DSS组小鼠血清IL-10及结肠MPO水平均与CON组有明显差异,地衣芽胞杆菌干预后有不同程度改变;使用非加权成对算术平均算法（UPGMA）对PCR-DGGE图谱中各泳道条带类型聚类分析结果显示各组小鼠结肠菌群结构具有显著差异,其中BL组增加双歧杆菌属Bifidobacterium saguini及疣微菌门的噬黏蛋白阿克曼菌（Akkermansia muciniphila）数量。结论 地衣芽胞杆菌可以有效改善DSS诱导的结肠炎症状,其作用机制与肠屏障功能和肠道菌群的调节有密切关系。     

不同途径下MIMP对 炎症性肠病小鼠肠道菌群结构的影响

目的通过葡聚糖硫酸钠诱导小鼠炎症性肠病(IBD)模型并观察不同途经下乳杆菌微小膜蛋白(MIMP)对炎症性肠病小鼠的紧密连接蛋白及菌群结构的影响。方法 C57BL/6小鼠24只根据DSS和MIMP不同干预组合将其分为4组:MIMP腹腔注射组(n=6)、MIMP灌胃组(n=6)、诱导肠炎组(n=6)和健康对照组(n=6),利用Western blot对各组小鼠肠道中紧密连接蛋白(Occludin、JAM-1和ZO-1)的表达进行检测,采用16SrRNA测序技术检测V4区鉴定细菌,并进行菌群差异分析。结果 MIMP腹腔注射及灌胃均可显著提高IBD小鼠肠道中紧密连接蛋白的表达水平,灌胃组效果更为显著;MIMP干预后小鼠肠道中拟杆菌门(Bacteroidetes)丰度增高、厚壁菌门(Firmicutes)及变形菌门(Proteobacteria)丰度降低,LEfSe分析、PCoA分析和PCA分析提示4组小鼠肠道菌群结构差异显著。结论不同途径下MIMP均可显著提高IBD小鼠肠道中紧密连接蛋白的表达水平,改善肠黏膜屏障功能,纠正小鼠肠道菌群结构紊乱。     

Effect of calcium on rat intestinal alkaline phosphatase activity and molecular aggregation

Two fractions of rat intestinal alkaline phosphatase (IAP) were detected by Western blot: 168 ± 6 and 475 ± 45 kDa. The low molecular weight fraction constitutes 43% of the isolated proteins exhibiting 82% of the enzymatic activity, and a heavier fraction constitutes 57% of the isolated proteins and has 18% of the enzymatic activity. Calcium produced an increase of the 475-kDa form to the detriment of the 168-kDa form. This work also describes the kinetic and structural changes of IAP as a function of calcium concentration. With [Ca²⁺] < 10 mmole/L, the Ca²⁺-IAP interaction fitted a binding model with 7.8 ± 4.4 moles of Ca²⁺ /mole of protein, affinity constant = 19.1 ± 8.4 L/mmole, and enzymatic activity increased as a linear function of [Ca²⁺] (r = 0.946 p < 0.01). On the other hand, with [Ca²⁺] >10 mmole/L the data did not fit this model and, the enzymatic activity decreased as a function of [Ca²⁺] (r = ? 0.703 p < 0.05).     

The intestinal microbiota: from health to disease,and back

Our understanding of the composition and the function of the intestinal microbiota has significantly increased over the past few years. In a series of reviews focusing on the role of the intestinal microbiota in health and disease, we explore recent conceptual and technological advances in this rapidly evolving research arena.     

Effect of adrenomedullin administration on acetic acid-induced colitis in rats

Adrenomedullin (AM) administered intracolonically ameliorated the severity of acetic acid-induced colonic ulceration in rats. Ulcers were induced by subserosal injection of acetic acid into the colon. AM-treated group was administered 0.25–1.0 μg of AM in 0.5 ml of saline intracolonically once a day; the control group received only saline. AM administration dose-dependently and significantly reduced the size of the ulcerative lesions, the associated edema, and the infiltration of the affected area by inflammatory cells. AM also reduced tissue levels of interleukin-6, but not interferon-γ. AM reduces the severity of acetic acid-induced colitis in rats, probably by inhibiting the production and/or release of Th-2 cell-derived factors such as interleukin-6.     

Involvement of intestinal alkaline phosphatase in serum apolipoprotein B-48 level and its association with ABO and secretor blood group types

Serum levels of intestinal alkaline phosphatase (IAP), a protein implicated in transcellular transport of chylomicrons, vary among ABO blood groups. In rat enterocytes, IAP is associated with chylomicron secretion, but the rat expresses only blood group A. It is not known whether chylomicron secretion may be affected in humans who express multiple blood group types. Serum samples from 40 healthy subjects were obtained after overnight fast and 3h after a high-fat meal, and assayed for IAP and apolipoprotein B-48 (apoB-48), both proteins exclusive to intestine, although only apoB-48 is found in chylomicrons. The two proteins were greater in subjects without blood antigen A (B and O) than in those with this antigen (A and AB); 2.4- and 4.7-fold for IAP and 1.5- and 2.0-fold for apoB-48 before and after the meal, respectively. Moreover, IAP and apoB-48 levels were strongly correlated in the subjects with the secretor phenotype (r > 0.81). These results indicate that IAP is strongly involved in chylomicron formation and fatty acid metabolism might change among ABO blood type. In addition, ABO blood type classification in apoB-48 measurement would improve the diagnostic value in the evaluation of metabolic syndrome.     

携带IL-10基因的双歧杆菌对溃疡性结肠炎小鼠肠道屏障功能的影响

目的 观察表达IL-10基因的双歧杆菌对溃疡性结肠炎（UC）小鼠肠道屏障功能的影响,进一步探讨转基因双歧杆菌治疗UC的相关机制。方法 25只小鼠分为空白对照组、结肠炎组（UC-blank组）、双歧杆菌治疗组（UC-bacteria组）、空质粒双歧杆菌治疗组（UC-pBBAD/X-bacteria组）和表达IL-10基因双歧杆菌治疗组（UC-BL-hIL-10-bacteria组）,每组5只。采用5% DSS诱导建立UC小鼠模型,利用前期研究已成功筛选出的可稳定表达hIL-10蛋白的BL-hIL-10菌株对小鼠进行治疗。采用HE染色评估小鼠结肠炎症情况;采用荧光定量PCR检测小鼠结肠组织Claudin1、2、4、5、7和8的表达水平;计算小鼠DAI。结果 （1）BL-hIL-10菌株可以降低UC小鼠DAI,减轻UC小鼠结肠组织炎症程度。（2）BL-hIL-10菌株能下调UC小鼠结肠组织Claudin2表达水平,同时能够上调Claudin1、4、5、7和8的表达水平。（3）UC-BL-hIL-10-bacteria组UC小鼠的治疗效果明显优于UC-bacteria组和UC-pBBAD/X-bacteria组。结论 BL-hIL-10菌株能改善UC小鼠结肠黏膜的通透性,其机制可能与其能上调Claudin1、4、5、7和8的表达水平,同时降低Claudin-2的表达水平有关。     

Autoantibodies against human tropomyosin isoform 5 in ulcerative colitis destroys colonic epithelial cells through antibody and complement-mediated lysis

INTRODUCTION: Patients with ulcerative colitis (UC) have IgG1 antibodies in serum and colon against human tropomyosin isoform 5 (hTM5), a cytoskeletal microfilament protein found intracellularly and on the surface of colonic epithelial cells (EC). These antibodies may be pathogenic in UC. METHODS: Sera from patients with UC (n=110) or Crohn's disease (CD) (n=50) and from healthy individuals (Hl) (n=30) were preincubated with recombinant hTM5 or bovine serum albumin (BSA), then cultured for 4h with (51)Cr-labelled colonic adenocarcinoma cells (LS180). Cytotoxicity was determined by (51)Cr release assay. RESULTS: All serum samples lysed up to 36% of LS180 cells regardless of the source of the serum. However, adding hTM5 to UC, but not to CD or HI, sera reduced cytotoxicity by up to 75%. This hTM5-induced inhibition of cytotoxicity was found especially with sera from UC patients with active disease, and was found even after total colectomy. The hTM5-induced inhibition was mediated by purified IgG from UC sera. Complement was involved since hTM5-induced inhibition of cytotoxicity declined with either heat inactivation of the sera or premixing sera with Fc fragments. CONCLUSIONS: This study shows that hTM5-specific IgG autoantibodies present in UC sera destroy LS180 cells by antibody and complement-mediated lysis. Such a phenomenon was not seen in CD or HI. This suggests an autoantigenic role of hTM5 and anti-hTM5 antibodies in the pathogenesis of UC. This observation may lead to novel diagnostic and therapeutic possibilities.     

植物乳杆菌ZDY2013与两歧双歧杆菌WBIN03对TNBS诱导小鼠结肠炎的缓解作用

目的 为阐明益生菌抗氧化与结肠炎的关系,对植物乳杆菌ZDY2013与两歧双歧杆菌WBIN03缓解三硝基苯磺酸（trinitro-benzene-sulfonic acid,TNBS）诱导的小鼠结肠炎进行探究。方法 通过对BALB/c小鼠肛门注射TNBS,构建小鼠结肠炎模型;分别采用植物乳杆菌ZDY2013与两歧双歧杆菌WBIN03的单菌悬液（109 CFU/mL）及1∶1混合菌悬液（109 CFU/mL）进行8 d灌胃治疗。结果 治疗组小鼠结肠组织炎性细胞浸润症状获得缓解,血清中谷胱甘肽过氧化物酶（glutathione peroxidase,GSH-PX）（t1=3.247,P1<0.05;t2=3.397,P2<0.05）、过氧化氢酶（catalase,CAT）（t1=5.289,P1<0.001;t2=3.563,P2<0.05）和总超氧化物歧化酶（total superoxide dismutase,T-SOD）（t1=3.317,P1<0.05;t2=3.551,P2<0.05）活性均有显著恢复。结论 植物乳杆菌ZDY2013与两歧双歧杆菌WBIN03可通过增强机体抗氧化酶活性,起到缓解TNBS诱导的小鼠结肠炎的作用。     

Transcriptional modulation of pattern recognition receptors in chronic colitis in mice is accompanied with Th1 and Th17 response

Pattern recognition receptors (PRRs) may contribute to inflammatory bowel diseases (IBD) development due to their microbial-sensing ability and the unique microenvironment in the inflamed gut. In this study, the PRR mRNA expression profile together with T cell-associated factors in the colon was examined using a chronic colitis mice model. 8–12 week old C57BL/6 mice were exposed to multiple dextran sodium sulfate (DSS) treatments interspersed with a rest period to mimic the course of chronic colitis. The clinical features and histological data were collected. The mRNA expressions of colonic PRRs, T cell-associated components were measured. Finally, the colons were scored for Foxp3+ cells. During chronic colitis, the histological data, but not the clinical manifestations demonstrated characteristic inflammatory symptoms in the distal colon. In contrast to acute colitis, the expression of all Toll-like receptors (Tlrs), except Tlr5 and Tlr9, was unaffected after repeated DSS treatments. The expression of Nod1 was decreased, while Nod2 increased. After third DSS treatment, only the expressions of Tlr3 and Tlr4 were significantly enhanced. Unlike other PRRs, decreased Tlr5 and increased Tlr9 mRNA expression persisted during the chronic colitis period. As the colitis progress, only the mRNA expression of Ifnγ and Il17 staid increased during chronic colitis, while the acute colitis-associated increase of Il23, and Il10 and Il12 was abolished. Finally, increased histological score of Foxp3+ cell in colon was found during the chronic colitis period. This study provides an expression pattern of PRRs during chronic colitis that is accompanied by a Th1- and Th17 cell-mediated immune response.     

整肠生对溃疡性结肠炎小鼠肠上皮屏障的保护作用

目的 探讨整肠生对溃疡性结肠炎小鼠肠道紧密连接蛋白表达以及对氧化应激反应的影响。方法 选用雄性8~10周龄C57BL/6小鼠40只,随机分为4组：对照组、模型组（3% DSS）、5-ASA组（3% DSS+5-ASA 200 mg/kg灌胃）和整肠生组（3% DSS+联合整肠生及5-ASA灌胃）,每组10只,造模7 d。观察各组小鼠便血程度、组织学损伤情况,通过投射电镜观察各组肠道上皮间紧密连接改变情况,应用Western blot和RT-PCR的方法,检测小鼠结肠黏膜紧密连接蛋白Occludin、ZO-1、Claudin-2的表达情况。结果 （1）与模型组比较,5-ASA组和整肠生组小鼠便血程度明显减轻,DAI评分显著降低（P<0.05）。整肠生组与5-ASA组比较,便血减轻,DAI评分降低（P<0.05）。（2）电镜显示,对照组肠上皮间紧密连接呈一条致密条带,结构完整,见细胞桥粒,微绒毛光滑、排列整齐,细胞间隙狭窄;模型组肠上皮间紧密连接结构松散、模糊、密度降低,桥粒结构消失,微绒毛稀疏,短缩且长短不一,细胞间隙增宽;各治疗组的紧密连接的破坏情况较模型组有不同程度的改善,整肠生组紧密连接清晰,细胞间隙缩窄,微绒毛排列整齐,出现细胞桥粒。（3）应用Western blot和Real time-PCR法检测,与正常组相比,模型组Occludin、ZO-1蛋白和mRNA表达显著下降,Claudin-2表达显著上调（P<0.05）;各治疗组较模型组Occludin、ZO-1蛋白表达上调,Claudin-2蛋白表达下调（P<0.05）,整肠生组较单用5-ASA组更明显提高Occludin、ZO-1蛋白和mRNA表达。（4）与正常组相比,模型组MDA含量增高,SOD活性降低,与5-ASA组相比,整肠生组能更显著地降低MDA含量,提高SOD活性（P<0.05）。结论 联合应用整肠生通过调节紧密连接蛋白Occludin、ZO-1的表达和降低氧化应激反应,来改善溃疡性结肠炎小鼠肠上皮屏障功能。     

Selective expression of detoxifying glutathione transferases in mouse colon: effect of experimental colitis and the presence of bacteria

Glutathione transferases (GSTs) play a central role in the cellular defense against harmful endogenous compounds and xenobiotics in mouse and man. The gastrointestinal channel is constantly exposed to bacteria, bacterial products, and xenobiotics. In the present study the distribution of alpha, mu, and pi class GSTs was examined immunohistologically in the colon of conventional and germ-free (GF) mice subjected to experimental colitis. The tissues samples were from conventional mice with and without colitis induced by dextran sulfate sodium (DSS); GF mice treated with DSS or carrageenan; and GF mice inoculated with normal mouse bacterial flora as well as with Lactobacillus GG. In conventional as well as in GF mice the mu and pi class GSTs showed reduced intestinal expression when colitis was induced. In contrast, the level of GSTs reacting with antibodies directed against the alpha class, in particular mGST A4-4, was elevated after induction of inflammation. Of special interest is mGST A4-4 because of its high catalytic activity with toxic products of lipid peroxidation. In the colon of conventionalized GF mice that were given mouse intestinal flora, the mGST A4-4 expression was increased with time for several weeks, but then showed a decrease to a normal level. Additionally, the inoculation of GF mice with Lactobacillus GG induced all the intestinal GSTs studied.     

Oligoelements in serum and intestinal tissue of pediatric IBD patients

IntroductionInflammatory bowel disease (IBD) develops through complex interplay of genetic, microbial, immune, and environmental factors. Trace elements alterations are commonly present in IBD and may have influence on IBD development. Heavy metal pollution is one of the major environmental issues nowadays and IBD incidence is rising in countries where industry starts to develop. Metals are implicated in processes that are connected to IBD pathogenesis.AimThe aim of this study was to investigate toxic and trace element levels in pediatric population of IBD patients both in serum and intestinal mucosa.Materials and methodsThis prospective study enrolled children newly diagnosed with IBD in University children’s hospital in Belgrade. Concentrations of thirteen elements: Al, As, Ca, Cd, Cr, Cu, Fe, K, Mg, Mn, Na, Se and Zn in serum and intestinal mucosa of 17 newly diagnosed children with IBD (10 Crohn’s disease (CD) and 7ulcerative colitis (UC)) and 10 controls were assessed using inductively coupled plasma mass spectrometry (ICP-MS). Intestinal mucosa samples were taken from terminal ileum and six different colon segments (cecum, ascending colon, colon transversum, descending and sigmoid colon and rectum).ResultsThe results demonstrated significant alterations in serum and intestinal mucosa concentrations of investigated elements. Serum iron was significantly decreased in IBD and CD group, compared to controls while serum Cu significantly differed between three investigated groups with highest concentration observed in CD children. Serum manganese was the highest in the UC subgroup. Terminal ileums of IBD patients contained significantly lower amount of Cu, Mg, Mn and Zn with Mn being significantly decreased also in CD patients compared to control. IBD patients’ caecum contained significantly less Mg and Cu while colon transversum tissue samples from IBD and Crohn’s patients contained significantly more chromium than controls. Moreover, sigmoid colon of IBD patients were poorer in Mg than controls (p < 0.05). Colon Al, As and Cd were significantly reduced in IBD, and UC children compared to control. Correlations of investigated elements in CD and UC groups were different from controls. Biochemical and clinical parameters showed correlation with element concentrations in intestines.ConclusionSera of CD, UC and control children significantly differ in Fe, Cu and Mn levels. Serum manganese was the highest in the UC subgroup creating the most prominent and only significant difference between UC and CD subgroups. Terminal ileum of IBD patients contained significantly lower amount of majority of investigated essential trace elements and toxic elements were significantly reduced in colon of IBD and UC patients. Investigation of macro- and microelement alterations in children and adults has potential to further elucidate IBD pathogenesis.     

红茶对新生幼鼠肠道菌群形成的影响

探讨红茶对新生幼鼠肠道菌群形成的影响,为从微生态角度研究红茶在人体肠道菌群形成过程中的作用机制奠定基础。

利用宏基因组测序技术检测4周龄新生幼鼠（4weeks组,n = 30）、12周龄正常对照组小鼠（control组,n = 15）和12周龄喂饲红茶水实验组小鼠（teadrink组,n = 15）的肠道菌群分布,分析3组样本的菌群差异情况,探求红茶对新生幼鼠肠道菌群形成的影响。

与control组相比,teadrink组小鼠肠道拟杆菌门（t = −7.711,P＜0.001）、拟杆菌科（t = −3.411,P = 0.009）、长尾嗤菌体科（t = −2.515,P = 0.036）、拟杆菌属（t = −2.693,P = 0.027）、邓肯菌属（t = −2.434,P = 0.041）、居海事城球杆菌属（t = −3.327,P = 0.029）、迪博邓肯菌（t = −2.679,P = 0.028）、普通居海事城球杆菌（t = −3.401,P = 0.027）和Duncaniella_sp._C9（t = −3.104,P = 0.035）相对丰度显著增加,厚壁菌门（t = 8.952,P＜0.001）、乳杆菌科（t = 13.102,P＜0.001）、消化链球菌科（t = 3.665,P = 0.021）、爱格菌科（t = 4.481,P = 0.002）、理研菌科（t = 3.626,P = 0.022）、乳杆菌属（t = 5.542,P = 0.004）、黏液乳杆菌属（t = 6.334,P = 0.002）、龙包茨菌属（t = 3.785,P = 0.005）、阿德勒菌属（t = 4.504,P = 0.002）、约氏乳杆菌（t = 4.282,P = 0.011）和罗伊氏粘液乳杆菌（t = 6.156,P = 0.003）相对丰度显著减少。与4weeks组相比,teadrink组小鼠肠道菌群分布差异大于control组。

红茶对新生幼鼠肠道菌群的形成能够产生影响,不同丰度的菌群可能通过调节碳水化合物代谢等途径来达到改善肠道菌群结构、增强肠道稳态的目的,具体代谢机制有待深入研究。

     

Kiwifruit extracts inhibit cytokine production by lipopolysaccharide-activated macrophages, and intestinal epithelial cells isolated from IL10 gene deficient mice

Inflammatory bowel disease (IBD) is a chronic, inflammatory disorder of the gastrointestinal tract involving an inappropriate immune response to commensal microorganisms in a genetically susceptible host. This study examined the effects of aqueous and ethyl acetate extracts of gold kiwifruit (Actinidia chinensis) or green kiwifruit (Actinidia deliciosa) using in vitro models of IBD. These models comprised primary macrophages and intestinal epithelial cells isolated from C57BL/5J and interleukin-10 gene deficient (Il10^−/−) mice and RAW 264.7, a murine macrophage-like cell line. All four kiwifruit extracts reduced the activation of these models after lipopolysaccharide stimulation, decreasing nitric oxide and cytokine secretion by both Il10^−/− and wild-type cells. The ethyl acetate extracts exhibited the highest anti-inflammatory activity, with almost complete suppression of lipopolysaccharide-stimulated macrophage activation. These results suggest that kiwifruit extracts have significant anti-inflammatory activity relevant to IBD. We suggest that the Il10^−/− mouse is a suitable model for further study of these compounds.