氨基酸对炎症性肠病的调控作用北大核心CSCD

炎症性肠病是胃肠道的一种慢性复发性炎症,包括克罗恩病和溃疡性结肠炎,患者众多,而且目前很难实现彻底治愈。由于患者消化功能受损,食物不容易吸收,很容易出现营养不良的情况,临床经常使用营养治疗来克服营养不足、改变炎症状态。氨基酸作为辅助营养治疗,可能有助于维持炎症性肠病患者的肠道完整性,减少炎症、氧化应激和肠道细胞死亡,对炎症性肠病的治疗起到积极的作用。最近在动物方面的研究已经证明氨基酸在炎症性肠病治疗中存在着巨大的潜力,氨基酸的供应和代谢可能是一种有前景的辅助治疗方法。本文就谷氨酰胺、精氨酸、甘氨酸等特定氨基酸的免疫调节作用进行综述,以期提供一种新的治疗炎症性肠病的思路。     

Bioactive dietary peptides and amino acids in inflammatory bowel disease

Inflammatory bowel disease (IBD), most commonly ulcerative colitis (UC) and Crohn’s disease (CD), is a chronic inflammation of the gastrointestinal tract. Patients affected with IBD experience symptoms including abdominal pain, persistent diarrhea, rectal bleeding, and weight loss. There is no cure for IBD; thus treatments typically focus on preventing complications, inducing and maintaining remission, and improving quality of life. During IBD, dysregulation of the intestinal immune system leads to increased production of pro-inflammatory cytokines, such as TNF-α and IL-6, and recruitment of activated immune cells to the intestine, causing tissue damage and perpetuating the inflammatory response. Recent biological therapies targeting specific inflammatory cytokines or pathways, in particular TNF-α, have shown promise, but not all patients respond to treatment, and some individuals become intolerant to treatment over time. Dietary peptides and amino acids (AAs) have been shown to modulate intestinal immune functions and influence inflammatory responses, and may be useful as alternative or ancillary treatments in IBD. This review focuses on dietary interventions for IBD treatment, in particular the role of dietary peptides and AAs in reducing inflammation, oxidative stress, and apoptosis in the gut, as well as recent advances in the cellular mechanisms responsible for their anti-inflammatory activity.     

Investigation of multiple susceptibility loci for inflammatory bowel disease in an Italian cohort of patients

Background

Recent GWAs and meta-analyses have outlined about 100 susceptibility genes/loci for inflammatory bowel diseases (IBD). In this study we aimed to investigate the influence of SNPs tagging the genes/loci PTGER4, TNFSF15, NKX2-3, ZNF365, IFNG, PTPN2, PSMG1, and HLA in a large pediatric- and adult-onset IBD Italian cohort.

Methods

Eight SNPs were assessed in 1,070 Crohn''s disease (CD), 1,213 ulcerative colitis (UC), 557 of whom being diagnosed at the age of ≤16 years, and 789 healthy controls. Correlations with sub-phenotypes and major variants of NOD2 gene were investigated.

Results

The SNPs tagging the TNFSF15, NKX2-3, ZNF365, and PTPN2 genes were associated with CD (P values ranging from 0.037 to 7×10⁻⁶). The SNPs tagging the PTGER4, NKX2-3, ZNF365, IFNG, PSMG1, and HLA area were associated with UC (P values 0.047 to 4×10⁻⁵). In the pediatric cohort the associations of TNFSF15, NKX2-3 with CD, and PTGER4, NKX2-3, ZNF365, IFNG, PSMG1 with UC, were confirmed. Association with TNFSF15 and pediatric UC was also reported. A correlation with NKX2-3 and need for surgery (P  =  0.038), and with HLA and steroid-responsiveness (P  =  0.024) in UC patients was observed. Moreover, significant association in our CD cohort with TNFSF15 SNP and colonic involvement (P  =  0.021), and with ZNF365 and ileal location (P  =  0.024) was demonstrated.

Conclusions

We confirmed in a large Italian cohort the associations with CD and UC of newly identified genes, both in adult and pediatric cohort of patients, with some influence on sub-phenotypes.     

Chemokine and cytokine levels in inflammatory bowel disease patients

Crohn’s disease (CD) and ulcerative colitis (UC), two forms of inflammatory bowel disease (IBD), are chronic, relapsing, and tissue destructive lesions that are accompanied by the uncontrolled activation of effector immune cells in the mucosa. Recent estimates indicate that there are 1.3 million annual cases of IBD in the United States, 50% of which consists of CD and 50% of UC. Chemokines and cytokines play a pivotal role in the regulation of mucosal inflammation by promoting leukocyte migration to sites of inflammation ultimately leading to tissue damage and destruction. In recent years, experimental studies in rodents have led to a better understanding of the role played by these inflammatory mediators in the development and progression of colitis. However, the clinical literature on IBD remains limited. Therefore, the aim of this study was to evaluate systemic concentrations of key chemokines and cytokines in forty-two IBD patients with a range of disease activity compared to levels found in ten healthy donors. We found a significant increase in an array of chemokines including macrophage migration factor (MIF), CCL25, CCL23, CXCL5, CXCL13, CXCL10, CXCL11, MCP1, and CCL21 in IBD patients as compared to normal healthy donors (P < 0.05). Further, we also report increases in the inflammatory cytokines IL-16, IFN-γ, IL-1β and TNF-α in IBD patients when compared to healthy donors (P < 0.05). These data clearly indicate an increase in circulating levels of specific chemokines and cytokines that are known to modulate systemic level through immune cells results in affecting local intestinal inflammation and tissue damage in IBD patients. Blockade of these inflammatory mediators should be explored as a mechanism to alleviate or even reverse symptoms of IBD.     

Extracellular release of free fatty acids by rat T lymphocytes is stimulus-dependent and is affected by dietary lipid manipulation

[(3)H]-Arachidonic acid-labelled rat T lymphocytes released radioactivity extracellularly when stimulated by the calcium ionophore A23187 or by monoclonal antibodies to some cell surface structures (CD2, CD5, CD11a, CD18, CD54, T-cell receptor) but not to others (CD49d, CD62L); release was greater with the calcium ionophore. Almost all of the radioactivity released from anti-CD2-stimulated lymphocytes was recovered in the free fatty acid fraction, whereas only about 50 per cent of that released after A23187 stimulation was recovered in this fraction. A23187 stimulation resulted in release of arachidonic acid from a variety of phospholipids (phosphatidylinositol, phosphatidylcholine and perhaps phosphatidylethanolamine), while the monoclonal antibody stimulation released arachidonic acid from phosphatidylinositol and perhaps phosphatidylcholine. Unstimulated lymphocytes released a range of fatty acids extracellularly, with palmitic acid accounting for 35-40 per cent and arachidonic acid for 5 per cent of released fatty acid. Stimulation of lymphocytes with either anti-CD2 or A23187 increased total fatty acid release 1.5- to 1.8-fold. In both cases palmitic acid remained the most predominant fatty acid released but the contribution of arachidonic acid increased. The type of lipid fed to the rats significantly influenced the amount and type of fatty acid released. Fish oil feeding significantly reduced extracellular fatty acid release by stimulated lymphocytes.     

Hydrocarbons,free fatty acids,and amino acids of sclerotia of Sclerotinia sclerotiorum

Summary Sclerotia of Sclerotinia sclerotiorum (Lib.) D By. were obtained from commercial pea-and bean-cleaning operations or grown on potato-dextrose agar and synthetic glucose-and sucrose-salts agar media. The crude fat (ether extract) content of sclerotia varied from 0.8 to 1.5%. Extraction and fractionation of the lipids followed by gas chromatographic analysis showed that sclerotia from pea cleanings contained one predominant hydrocarbon which was absent from sclerotia produced in the laboratory. Sclerotia from natural sources and grown in the laboratory contained a similar distribution of C₁₈ unsaturated free fatty acids, however, quantitative differences were noted. Palmitic, oleic and linoleic were the major free fatty acids of the laboratory-grown sclerotia while a high proportion of linoleic acid was also found in sclerotia from natural sources. Sclerotia were fractionated into water-soluble and water-insoluble fractions. After acid hydrolysis of the waterinsoluble fraction, both fractions were analyzed for amino acids. Twenty-one compounds, including 2 unknowns, were detected in the soluble fraction. The hydrolyzates contained 19 amino acids, including the same 2 unknowns. Two compounds tentatively identified as ornithine and -aminobutyric acid were found only in the water-soluble fraction. The relative amino acid composition of the water-insoluble fraction of sclerotia from various sources was fairly constant but the arginine content decreased on the synthetic media.     

Protein and free amino acids in wheat grain as affected by soil types and salinity levels in irrigation water

Summary The effects of four lysimeter soil series under three salinity levels were evaluated for grain yield, wt/1000 seeds, protein, and amino acids in Mexican dwarf wheat (Triticum aestivum L. var. Cajeme 71). The soil series consisted of: Holtville clay loam, Greenfield sandy loam, San Emigdio sandy loam, and Altamont clay loam. The irrigation water salinity levels were designated: low –2.2 mmho, medium –4.2 mmho, and high –7.1 mmho.No significant differences were found in the amount of grain harvested or wt/1000 seeds in the 1976 crop produced on the differential soil series. The yield of the 1977 crop was significantly affected by the soil types.Effects of soil type on the protein amino acids in the grain in both years were similar. Significantly higher protein amino acid levels of histidine, arginine, aspartic acid, threonine, serine, glutamic acid, glycine, alanine, cystine, valine, methionine, isoleucine, leucine, tyrosine, and phenylalanine were found in the grain grown on Altamont clay loam soil than the other types.The free amino acids in grain from the 1976 and 1977 crops were similarly affected by the soil types, except that the quantitative values of the free amino acids were substantially lower in 1977 than in 1976. The free amino acids significantly influenced by soil types were tryptophane, lysine, arginine, aspartic acid, threonine, serine, glycine, alanine, valine, isoleucine, tyrosine, and phenylalanine. In both years' crops, the sum of the free amino acid fractions was significantly higher in the grain produced on the Altamont soil than on the other soils.Salinity level in the irrigation water did not affect the 1976 crop yield or wt/1000 seeds. Although yields of the 1977 crop were significantly reduced by salinity, the wt/1000 seeds was not. The sum of protein amino acids was significantly higher in the 1976 and 1977 grain crops irrigated with high salinity water than in low salinity irrigated crops.An increased salinity irrigation water significantly reduced the sum of free amino acid fractions in the 1976 grain crop. Since some of the free amino acids in the 1977 grain crop increased while the others decreased due to the salinity level in the irrigation water, the sum of the free amino acid fractions was not significantly influenced.Significant interactions were found between soil types and salinity levels on free arginine, threonine, serine, glutamic acid, and alanine, and also on the sum of the free amino acids in the 1976 wheat grain. In the 1977 wheat grain, there were significant interactions between soil types and salinity levels on the free glutamic acid, valine, leucine, tyrosine, and phenylalanine, and on protein serine, glutamic acid, glycine, alanine, and the sum of the protein amino acids.The amounts of essential amino acids expressed as mg of amino acid/g of protein were not affected by the soil types or salinity levels. With the exception of lysine, and possibly threonine and methionine plus cystine, the essential amino acids were present in the grain at concentrations equal to or greater than recommended by WHO and FAO.     

Mesenchymal stromal cell function is not affected by drugs used in the treatment of inflammatory bowel disease

Background and aimsMesenchymal stromal cells (MSC) have both multilineage differentiation capacity and immunosuppressive properties. Promising results with MSC administration have been obtained in experimental colitis. Clinical application of MSC for the treatment of inflammatory bowel disease (IBD) is currently under investigation in phase I–III trials in patients with past or concurrent immunomodulating therapy. However, little is known about MSC interactions with these immunosuppressive drugs. To address this issue we studied the combined effect of MSC and IBD drugs in in vitro functionality assaysMethodsThe effects of azathioprine, methotrexate, 6-mercaptopurine and anti-tumor necrosis factor (TNF)-α on MSC phenotype, survival, differentiation capacity and immunosuppressive capacity were studiedResultsMSC exposed to physiologically relevant concentrations of IBD drugs displayed a normal morphology and fulfilled phenotypic and functional criteria for MSC. Differentiation into adipocyte and osteocyte lineages was not affected and cells exhibited normal survival after exposure to the various drugs. MSC suppression of peripheral blood mononuclear cell (PBMC) proliferation in vitro was not hampered by IBD drugs. In fact, in the presence of 6-mercaptopurine and anti-TNF-α antibodies, the inhibitory effect of this drug alone was enhanced, suggesting an additive effect of pharmacotherapy and MSC treatmentConclusionsThis study demonstrates that, in vitro, MSC phenotype and function are not affected by therapeutic concentrations of drugs commonly used in the treatment of IBD. These findings are important for the potential clinical use of MSC in combination with immunomodulating drugs and anti-TNF-α therapy.     

Sterols, fatty acids and free amino acids from two Helvella species

Sterols, fatty acids and free amino acids of Helvella crispa and H. monachella were investigated. They contained traces of ergosterol and a high amount of ergosta 5.22-dien-3β-ol. Linoleic acid and l-Dopa are the most abundant fatty acid and free amino acid, respectively. The aqueous extract of H. monachella inhibits prostaglandin release by rat peritoneal leucocyres in vitro.     

Purine,kynurenine, neopterin and lipid peroxidation levels in inflammatory bowel disease

The kynurenine metabolites of tryptophan may be involved in the regulation of neuronal activity and thus gut motility and secretion. We have now performed a pilot study to measure serum concentrations of purines and kynurenines in patients with mild inflammatory bowel disease, as well as in sex- and age-matched control subjects. For some analyses, the patients were subdivided into subgroups of those with Crohn's disease and those with ulcerative colitis. The analyses indicated an increased activity in one branch of the kynurenine pathway. While there was no demonstrable difference in neopterin levels in either of the patient groups compared with controls, indicating that the disorders were in an inactive quiescent phase, both groups showed significantly higher levels of lipid peroxidation products. This suggests the presence of increased oxidative stress even during relative disease inactivity. The increased level of kynurenic acid may represent either a compensatory response to elevated activation of enteric neurones or a primary abnormality which induces a compensatory increase in gut activity. In either case, the data may indicate a role for kynurenine modulation of glutamate receptors in the symptoms of inflammatory bowel disease.     

Free radical-mediated oxidation of free amino acids and amino acid residues in proteins

Summary. We summarize here results of studies designed to elucidate basic mechanisms of reactive oxygen (ROS)-mediated oxidation of proteins and free amino acids. These studies have shown that oxidation of proteins can lead to hydroxylation of aromatic groups and aliphatic amino acid side chains, nitration of aromatic amino acid residues, nitrosylation of sulfhydryl groups, sulfoxidation of methionine residues, chlorination of aromatic groups and primary amino groups, and to conversion of some amino acid residues to carbonyl derivatives. Oxidation can lead also to cleavage of the polypeptide chain and to formation of cross-linked protein aggregates. Furthermore, functional groups of proteins can react with oxidation products of polyunsaturated fatty acids and with carbohydrate derivatives (glycation/glycoxidation) to produce inactive derivatives. Highly specific methods have been developed for the detection and assay of the various kinds of protein modifications. Because the generation of carbonyl derivatives occurs by many different mechanisms, the level of carbonyl groups in proteins is widely used as a marker of oxidative protein damage. The level of oxidized proteins increases with aging and in a number of age-related diseases. However, the accumulation of oxidized protein is a complex function of the rates of ROS formation, antioxidant levels, and the ability to proteolytically eliminate oxidized forms of proteins. Thus, the accumulation of oxidized proteins is also dependent upon genetic factors and individual life styles. It is noteworthy that surface-exposed methionine and cysteine residues of proteins are particularly sensitive to oxidation by almost all forms of ROS; however, unlike other kinds of oxidation the oxidation of these sulfur-containing amino acid residues is reversible. It is thus evident that the cyclic oxidation and reduction of the sulfur-containing amino acids may serve as an important antioxidant mechanism, and also that these reversible oxidations may provide an important mechanism for the regulation of some enzyme functions.     

Serotoninergic neurotransmission is affected by n-3 polyunsaturated fatty acids in the rat

We explored the effects of chronic alpha-linolenic acid dietary deficiency on serotoninergic neurotransmission. In vivo synaptic serotonin (5-HT) levels were studied in basal and pharmacologically stimulated conditions using intracerebral microdialysis in the hippocampus of awake 2-month-old rats. We also studied the effects of reversion of the deficient diet on fatty acid composition and serotoninergic neurotransmission. A balanced (control) diet was supplied to deficient rats at different stages of development, i.e. from birth, 7, 14 or 21 days of age. We demonstrated that chronic n-3 polyunsaturated fatty acid dietary deficiency induced changes in the synaptic levels of 5-HT both in basal conditions and after pharmacological stimulation with fenfluramine. Higher levels of basal 5-HT release and lower levels of 5-HT-stimulated release were found in deficient than in control rats. These neurochemical modifications were reversed by supply of the balanced diet provided at birth or during the first 2 weeks of life through the maternal milk, whereas they persisted if the balanced diet was given from weaning (at 3 weeks of age). This suggests that provision of essential fatty acids is durably able to affect brain function and that this is related to the developmental stage during which the deficiency occurs.     

Fatty acid transport in adipocytes monitored by imaging intracellular free fatty acid levels

Transport of free fatty acids (FFA) across the adipocyte plasma membrane is critical for maintaining homeostasis. To determine the membrane's role in regulating transport we describe here the first measurements of the intracellular (unbound) FFA concentration ([FFA(i)]) and their use in monitoring transport of FFA across 3T3F442A adipocytes. [FFA(i)] was measured by microinjecting cells with ADIFAB, a fluorescently labeled fatty acid-binding protein that is used to measure unbound FFA levels. We used ratio fluorescence microscopy of intracellular ADIFAB to image unbound FFA levels and determined the time course of [FFA(i)] in response to changing the extracellular unbound FFA concentration ([FFA(o)]). [FFA(o)] was clamped at defined levels using complexes of FFA and bovine serum albumin. We show that FFA influx is slow, requiring about 300 s to reach steady state (rate constant approximately 0.02 s(-1)) and saturable (K(o) approximately 200 nm). Efflux is twice as fast as influx, for zero [FFA(o)], but decreases with increasing [FFA(o)]. Surprisingly, at steady state [FFA(i)] is 2-5-fold (average 2-fold) greater than [FFA(o)] and this [FFA(i)]/[FFA(o)] gradient is abolished by depleting cellular ATP. Our results indicate that FFA transport across adipocyte membranes is highly regulated, involving an ATP-driven pump and a mechanism for gating efflux that is sensitive to [FFA(o)]. These characteristics are well described by a membrane carrier model but are not consistent with FFA transport across the membrane's lipid phase. We suggest that these characteristics are important in regulating circulating FFA levels by the adipocyte.     

Volatile fatty and dissolved free amino acids in Organic Lake,Vestfold Hills,East Antarctica

The distributions of volatile fatty acids (C1–C5) (VFAs) and dissolved free amino acids (DFAAs) in Organic Lake, a shallow hypersaline meromictic lake in the Vestfold Hills, East Antarctica, were determined during the winter and spring of 1987. The concentrations of the two VFAs detected, formic and acetic acid, were low (<10 M) in the oxic upper waters of the lake, but increased dramatically (up to 250M) beneath the oxycline. The distribution of DFAAs was similar, with a total concentration of 280M in the anoxic water early in the study. The amino acids occurring at the highest concentrations were alanine, glutamic acid, leucine and valine. Total VFA concentrations in the anoxic zone increased during the study period, but the total amino acid concentration dropped significantly to 64 M by the end of the study. The high concentrations beneath the oxycline were probably the result of bacterial utilisation of these substrates that was slow due to the high salinity and low temperature of the lake water.