基质金属蛋白酶-1、基质金属蛋白酶-2和基质金属蛋白酶-3在溃疡性结肠炎患者肠黏膜的表达

目的了解部分基质金属蛋白酶(MMPs)在溃疡性结肠炎(UC)患者肠黏膜的表达,为明确MMPs在UC发病机制的作用奠定基础。方法肠镜下取25例UC患者的病变肠黏膜及20例对照组肠黏膜,采用逆转录多聚酶链反应(RT-PCR)检测基质金属蛋白酶-1(MMP-1)、基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-3(MMP-3)和金属蛋白酶抑制剂(TIMP-1)mRNAs的表达量。结果UC患者肠黏膜的MMP-1、MMP-2和MMP-3 mRNAs的表达量分别为0.37±0.19、0.40±0.16和0.24±0.13,而对照组肠黏膜的表达分别为0.18±0.15、0.16±0.13和0.09±0.13,二组比较均有显著性差异(P<0.05)。结论溃疡性结肠炎的炎症性病变肠黏膜中,基质金属蛋白酶尤其是MMP-1和MMP-3的表达均明显高于正常的肠黏膜,提示MMPs在UC的发病机制中起重要作用。     

MicroRNA exhibit altered expression in the inflamed colonic mucosa of ulcerative colitis patients

AIM To investigate the miRNA expression in colonic mucosal biopsies from endoscopically inflamed and non inflamed regions of ulcerative colitis(UC) patients. METHODS Colonic mucosal pinch biopsies were analyzed from the inflamed and non inflamed regions of same UC patient. Total RNA was isolated and differential miRNA profiling was done using microarray platform. Quantitative Real Time PCR was performed in colonic biopsies from inflamed(n = 8) and non-inflamed(n = 8) regions of UC and controls(n = 8) to validate the differential expression of miRNA. Potential targets of dysregulated miRNA were identified by using in silico prediction tools and probable role of these miRNA in inflammatory pathways were predicted.RESULTS The miRNA profile of inflamed colonic mucosa differs significantly from the non-inflamed. Real time PCR analysis showed that some of the miRNA were differentially expressed in the inflamed mucosa as compared to non inflamed mucosa and controls(mi R-125 b, mi R-223, miR-138, and mi R-155), while(miR-200a) did not show any significant changes. In contrast to microarray, where mi R-378 d showed downregulation in the inflamed mucosa, q RT-PCR showed a significant upregulation in the inflamed mucosa as compared to the non inflamed. The in silico prediction analysis revealed that the genes targeted by these mi RNAs play role in the major signaling pathways like MAPK pathway, NF-κB signaling pathway, cell adhesion molecules which are all assciated with UC.CONCLUSION The present study reports disease specific alteration in the expression of mi R-125 b, mi R-155, mi R-223 and mi R-138 in UC patients and also predict their biological significance.     

Telomere shortening in the colonic mucosa of patients with ulcerative colitis

Telomere length in human somatic cells gradually decreases with the number of cell divisions and is regarded as a marker of somatic cell turnover. Mucosal cells of the affected colon show rapid turnover in individuals with active ulcerative colitis (UC). Telomere length was determined by Southern blot analysis of terminal restriction fragments (TRFs) from the colonic mucosa of 17 patients with UC in remission, two of whom showed dysplasia, and 17 control subjects without colitis. For each individual, mean TRF length was compared between rectal mucosa and unaffected cecal mucosa. The mean TRF length of the rectal mucosa was significantly less than that of cecal mucosa in UC patients (7.87 ± 0.36 kb versus 8.77 ± 0.21 kb; P = 0.0015, Wilcoxon signed rank test), whereas no significant difference was detected in the control subjects. The extent of telomere shortening was 10.6 ± 3.35% in UC patients, compared with 0.8 ± 0.64% in noncolitis controls (P = 0.0024, Mann-Whitney U-test). Four UC patients, two of whom had dysplasia, showed telomere shortening of more than 20% in the rectal mucosa. These observations suggest that telomere shortening in the colonic mucosa of individuals with UC may represent the history of mucosal inflammation during disease of long duration, and that it may contribute to aneuploidy in UC. (Received May 6, 1997; accepted Sept. 26, 1997)     

Urocortin 1 in colonic mucosa in patients with ulcerative colitis

Ulcerative colitis (UC) is characterized by a long-standing chronic inflammation of the bowel with intermittent periods of exacerbation and remission. Its acute exacerbation appears to be related to various stresses. Urocortin 1 (Ucn1) may play important roles in integrated local responses to stress. We therefore examined local production of Ucn1 in patients with UC by immunohistochemistry and mRNA in situ hybridization. Ucn1 immunoreactivity was predominantly detected in lamina propria plasma cells and enterochromaffin cells. In UC patients without glucocorticoid treatment, Ucn1-positive cells and plasma cells increased in proportion to the severity of inflammation (P < 0.0001). Ucn1-positive cells significantly increased in UC patients with advanced inflammatory grades, compared with a control group (P < 0.0001) and nonspecific colitis group (P < 0.0001). In glucocorticoid-treated patients, Ucn1-positive cells were significantly lower in number, compared with the nonglucocorticoid-treated group. Ucn1 mRNA was expressed in lamina propria plasma cells, and both corticotropin-releasing factor(1) and corticotropin-releasing factor(2(a)) mRNAs were also partially coexpressed in these cells and macrophages. The present study showed that Ucn1-positive cells were correlated with the severity of inflammation in colonic mucosa with UC, and glucocorticoid treatment decreased these cells. Ucn1 therefore may act as a possible local immune-inflammatory mediator in UC.     

溃疡性结肠炎结肠黏膜脱氧核糖核酸甲基转移酶3a表达及作用研究

目的 探讨溃疡性结肠炎(UC)患者结肠黏膜固有层内单个核细胞中脱氧核糖核酸甲基转移酶3a(DNMT3a)与白细胞介素-4(IL-4)、γ-干扰素(IFN-γ)表达的相关性.方法 选取2009年11月至2010年3月天津医科大学总医院60例结肠内镜活检标本,其中活动期UC和正常对照组各30例.应用免疫组织化学染色法(S...     

Early changes of ileoanal pouch mucosa in patients with ulcerative colitis

BACKGROUND/AIMS: Following closure of the protective ileostomy the mucosa of ileoanal pouches undergoes characteristic changes, which are thought to be caused by the new luminal environment Conventional histopathology and immunohistochemical markers were compared in serial pouch biopsies from patients with ulcerative colitis (UC) in the early period of function. METHODS: Biopsy specimens were obtained from nine patients during ileostomy closure, 24 h after the first bowel action, then 6 days, 6 weeks and 3 months postoperatively. Cryostat sections were stained with haematoxylin and eosin (H&E) for assessment of mucosal inflammation and morphometry, and for lymphocyte subtypes (CD3, CD4, CD8), macrophages (CD68), common leucocyte antigen (CD45), and HLA-DR, using a three-stage immunoperoxidase reaction. RESULTS: Within 5 days of pouch function the index for mucosal atrophy (villous height/total mucosal thickness ratio) decreased significantly from a median (range) of 0.66 (0.39-0.69) to 0.52 (0.37-0.61)(P = 0.02). Intraepithelial lymphocyte counts diminished from 10.1 (5.6-21.1) to 7.0 (2.2-8.2)(P = 0.007) per 100 epithelial cells and remained so. In the lamina propria, apart from a transient increase in CD3 positive cells at 5 days (from 92.7 (58-165) to 100.5 (57-234) per unit area; P = 0.038), no significant changes were observed. H&E grading and HLA-DR expression remained unchanged. CONCLUSIONS: While there was no significant change in mucosal morphology and mucosal leucocyte subpopulations after 24 h of pouch function, the ileal pouch mucosa in UC patients had responded significantly within 5 days. Further studies into cell function and signalling are required.     

Low detoxification capacity in the ileal pouch mucosa of patients with ulcerative colitis

BACKGROUND: An ileal pouch-anal anastomosis has become the most widely accepted procedure for surgical treatment of patients with ulcerative colitis (UC). The primary function of the ileum within the pouch changes from absorption to storage. Malignancies have been described in the pouch mucosa. The detoxifying glutathione S-transferase (GST) enzymes are involved in the mucosal protection against toxins and carcinogens. Levels of GSTs are much higher in the ileum as compared with the colon. The adaptation of the ileal pouch mucosa into a more colon-like phenotype possibly influences the activity and levels of GST. This study compares the detoxification capacity of GST of the afferent ileal limb mucosa with the ileal pouch mucosa of patients with UC. METHODS: Biopsies from normal-appearing mucosa from the ileal pouch and the ileal afferent limb were obtained from 18 patients with UC. GST isoforms were quantified by immunoblotting. GST activity was measured spectrophotometrically, and glutathione and cysteine levels were determined by high-performance liquid chromatography. RESULTS: The GST activity and GSTA1+A2 levels were significantly lower in the pouch compared with the afferent ileal limb of patients with UC, whereas the GSTP1 levels were higher in the pouch. No differences were observed in the levels of GSTM1, GSTT1, glutathione, or cysteine. CONCLUSIONS: The lower GST detoxification activity in the pouch mucosa of patients with UC may result in higher levels of toxins and carcinogens and thus partly contribute to the risk of developing malignancies in the pouch.     

溃疡性结肠炎患者核因子-κB活化与细胞因子基因表达

目的　探讨溃疡性结肠炎 (UC)患者肠黏膜活检组织细胞因子mRNA的表达及其与NF κB活化的关系 ,以及抗炎药物 (柳氮磺吡啶和糖皮质激素 )对其的影响。方法　 31例来自四川大学华西医院的UC患者 (符合 1993年太原会议制定的UC诊断标准 )被纳入本研究。其中 17例使用过药物 (柳氮磺吡啶或柳氮磺吡啶 +糖皮质激素 )治疗 ,14例未用过任何与UC治疗相关的药物 ,11例同期结肠癌患者 (取其癌旁正常组织 )作为对照。采用 :(1)凝胶电泳迁移率改变分析检测核因子 (NF) κBDNA结合活性 ;(2 )逆转录聚合酶链反应检测白细胞介素 (IL) 1βmRNA和IL 8mRNA的表达。 结果　 (1)UC患者肠黏膜活检组织IL 1βmRNA和IL 8mRNA表达与对照组相比明显升高 (P <0 0 5 ) ,且与NF κBDNA结合活性呈显著正相关 (IL 1β :r=0 836 3,P <0 0 5 ;IL 8:r=0 6 0 2 4 ,P <0 0 5 )。 (2 )糖皮质激素和柳氮磺吡啶明显抑制NF κB的活性 ,降低IL 1βmRNA和IL 8mRNA的表达。结论　 (1)NF κB是UC细胞因子释放的关键调控因素 ,在UC的发生和发展中起着十分重要的作用。 (2 )糖皮质激素和柳氮磺吡啶可能通过抑制NF κB的活性 ,减少细胞因子的表达而起到抗炎作用。     

PPAR-γ在溃疡性结肠炎粘膜中的表达

目的了解过氧化物酶增生激活受体（ＰＰＡＲ－γ）在溃疡性结肠炎（ＵＣ）患者结肠活检粘膜中的表达及其与疾病严重度的关系。方法　采用免疫组化法测定３１例ＵＣ患者和１２例对照组的结肠活检粘膜中ＰＰＡＲ－γ的表达情况。结果　ＵＣ患者受累及相对正常的结肠上皮ＰＰＡＲ－γ的表达均较正常对照组下降，三组分别为２４．７％±１．４９％、４３．８％±２．０３％、７０．２％±３．７５％（Ｐ＜０．０５），且病变越重，表达越低。结论ＵＣ患者内镜下活检粘膜中ＰＰＡＲ－γ的表达下降，并与疾病严重程度成负相关，提示其可能与ＵＣ的发病有关。     

Corticotropin releasing hormone in colonic mucosa in patients with ulcerative colitis.

Corticotropin releasing hormone (CRH) is a key hormone in integrated response to stress, acting as the major regulator of the hypothalamic-pituitary-adrenal axis. Recently, local production of CRH has been detected in normal human colonic enterochromaffin cells. CRH is locally secreted in granulomatous and arthritic tissues in rats and humans, where it seems to act as a local proinflammatory agent. To find out if CRH is present in colonic tissues of patients with ulcerative colitis, this study examined the expression of this peptide in the large bowel of patients with ulcerative colitis. Colonic tissues of patients with ulcerative colitis obtained by endoscopic biopsy were immunostained with anti-CRH antibody. CRH messenger (m) RNA was also examined in biopsy specimens of ulcerative colitis by the reverse transcribed polymerase chain reaction method and by in situ hybridisation. Considerably enhanced expression of immunoreactive CRH was found in mucosal inflammatory cells. Intense staining with anti-CRH antibody was also shown in mucosal macrophages. CRH mRNA was expressed in mucosal epithelial cells. The expression of immunoreactive CRH in colonic mucosal epithelial cells of ulcerative colitis slightly increased, but not significantly, compared with normal colonic mucosal epithelial cells. These results suggest that CRH may play a part in the modulation of intestinal immune and inflammatory system, and as a modulator in the pathogenesis of ulcerative colitis.     

Endoscopic finding of spontaneous hemorrhage correlates with tumor necrosis factor alpha expression in colonic mucosa of patients with ulcerative colitis

Background

Ulcerative colitis (UC) is an intractable colonic disease, and it shows several endoscopic findings. Recently, it was reported that the expression level of mucosal tumor necrosis factor alpha (TNF-α) was useful for predicting patient response to infliximab. However, no data regarding the value of endoscopic findings to predict treatment efficacy or cytokine expression level exist.

Objective

We investigated the expression of leukocyte adhesion-related molecules and cytokines in colonic mucosa and compared it to endoscopic findings.

Methods

One hundred and fifty-nine patients were enrolled. Tissue samples were obtained by colonic biopsy from patients with UC. Colitis activity was determined by Matts’ criteria. The degree of mRNA expression of TNF-α, interferon gamma (IFN-γ), interleukin (IL)-8, IL-17A, and mucosal vascular addressin adhesion molecule-1 (MAdCAM-1) in mucosal samples was determined by real-time quantitative polymerase chain reaction. These expression levels were compared with the degree of Matts’ grade and individual endoscopic findings.

Results

The expression of TNF-α, IFN-γ, IL-8, IL-17A, and MAdCAM-1 mRNA significantly increased as Matts’ endoscopic grade elevated. Actively inflamed mucosa with spontaneous hemorrhage revealed a significantly increased expression level of TNF-α mRNA than that without spontaneous hemorrhage. No other individual endoscopic parameter was significantly correlated with the expression level of TNF-α mRNA.

Conclusions

Inflamed mucosa with spontaneous hemorrhage may suggest increased expression of TNF-α mRNA levels in colonic mucosa of UC patients, which could predict a lower response to infliximab treatment and more aggressive induction regime or change to other therapy should be taken into account.     

感染后肠易激综合征与溃疡性结肠炎缓解期患者肠黏膜细胞因子表达的相关性

目的:探讨感染后肠易激综合征(PI-IBS)与溃疡性结肠炎(UC)缓解期患者结肠黏膜细胞因子表达的相关性.方法:PI-IBS组26例,UC组45例及对照组30例,结肠镜下活检降结肠和直肠黏膜标本,采用免疫组化SABC法检测其肠黏膜P物质(SP)与IL-2,IFN-γ的表达情况.结果:PI-IBS组降结肠和直肠黏膜IFN-γ和IL-2阳性率表达、SP强度均值和面积高于对照组(IFN-γ:χ2=13.781,14.012,P＜0.01;IL-2:χ2=13.890,13.931,P＜0.01;SP强度:t=3.623,3.722,P＜0.01;SP面积:t=3.454,3.561,P＜0.01),但与UC组患者无显著差异.降结肠、直肠黏膜IFN-γ,IL-2阳性表达的PI-IBS患者,SP强度均值(t=2.202,2.220,P＜0.05)、面积高于对照组(t=2.301,2.252,P＜0.05),与UC组患者也无显著差异.结论:PI-IBS和UC缓解期患者细胞因子表达无显著差异.从神经-免疫机制上分析认为IBS与炎症性肠病(IBD)之间存在某种相关性,IBS可能是轻微的IBD.     

Differential expression of CCR5 and CRTH2 on infiltrated cells in colonic mucosa of patients with ulcerative colitis

BACKGROUND AND AIM: The pathogenesis of ulcerative colitis (UC) is unclear, but abnormal infiltration of T lymphocytes in the colonic mucosa has been implicated in the mucosal tissue damage. The abnormal cytokine production because of a T helper (h)1/Th2 imbalance may play an important role in continuing inflammation in the colonic mucosa. In the present study, the expression of chemokine receptor 5 (CCR5) as a Th1 marker and a chemoattractant receptor-homologs molecule expressed on Th2 cells (CRTH2) were investigated in order to analyze impaired Th1/Th2 responses in the colonic mucosa of UC patients. METHODS: Tissue samples were obtained by colonic biopsies from patients with UC or colonic polyps, with informed consent. Immunohistochemical analysis was performed on periodate, lysine-paraformaldehyde-fixed serial cryostat sections using the labeled streptavidin biotin method. Monoclonal antibodies against CD4, CCR5 or CRTH2 were used as primary antibodies. The number of cells expressing CD4, CCR5 or CRTH2 per unit area was calculated by using an image analyzer. RESULTS: In the patients with UC, the numbers of CD4- and CCR5-positive cells were significantly increased in inflamed mucosa, and appeared to be correlated with the disease activity. The infiltration of CRTH2-positive cells was predominantly observed in the mildly inflamed or the margin of inflamed mucosa of UC patients. CONCLUSION: There is a possibility that Th1 responses significantly occur in colonic mucosa with severe inflammation, while Th2 responses mainly occur with mild inflammation in UC patients. The Th1/Th2 imbalance in colonic mucosa may be related to the disease progression of UC.     

Changes in pulmonary function in patients with ulcerative colitis

OBJECTIVES: Information on the occurrence and frequency of pulmonary involvement in patients with ulcerative colitis (UC) is inconsistent. Some authors reported pulmonary impairment with UC by standard pulmonary function tests (PFTs) and documented a reduced diffusing capacity for carbon monoxide (DLCO) especially in patients with active disease, whereas others could not detect differences in routine PFTs between UC patients and controls. AIM: The aim of this prospective study was to determine the frequency and type of pulmonary dysfunction in patients with UC with respect to disease activity. Furthermore, to evaluate the influence of smoking, nutritional status, sputum cytology and sulphasalazine therapy on PFT parameters. PATIENTS AND METHODS: Twenty-six patients with UC (20 with active disease, 6 inactive) and 16 age and sex matched healthy controls were investigated with respect to the following pulmonary function tests, forced vital capacity (FVC), forced expiratory volume in the 1s (FEV(1)%) and their ratio (FEV(1)/FVC) and forced expiratory flow 25-75% (FEF25-75%) as well as oxygen saturation. For UC patients, colonoscopy and biopsy were done. Disease activity was assessed by Truelove index for UC. Induced sputum was sampled for cytology. Smoking habit, body mass index (BMI) and medications were recorded. RESULTS: Fifteen out of 26 patients with UC (57.6%) exhibited at least one pathological pulmonary function test (<80% of predicted value). Small airway obstruction was reported in the 15 patients, restrictive dysfunction in 30.7% and obstructive dysfunction in 11.5%. The impairment of PFTs was significant and more pronounced in patients with active disease, FVC (-14% of predicted), FEV(1) (-9% of predicted) and FEF25-75% (-32% of predicted), P<0.01, 0.05 and 0.01, respectively. There was no significant influence of smoking and medications on PFTs. CONCLUSIONS: UC patients show significantly decreased lung function tests in comparison to healthy controls. The impairment in active disease exceeded that during the remission. Early recognition is important, as they can be strikingly steroid responsive.     

双歧杆菌影响溃疡性结肠炎患者肠黏膜的研究

目的　研究双歧杆菌对溃疡性结肠炎 (UC)患者肠黏膜内核因子κB (NF κB)的表达和激活的影响及肿瘤坏死因子 (TNF) α、白细胞介素 (IL) 1β、IL 10mRNA表达的影响。 方法　 30例UC患者经治疗后 ,应用Western蛋白印迹分析检测NF κBp6 5表达情况 ;用凝胶电泳迁移率改变分析检测NF κB与DNA结合活性 ;逆转录 PCR检测肠黏膜内细胞因子的表达。结果　应用双歧杆菌后 3例复发 ,对照组 14例复发 ,与对照组相比 ,NF κB与DNA结合活性明显降低 ,蛋白表达量明显下降 (治疗前 0 82± 0 0 5 ,治疗后 0 31± 0 0 5 ,P <0 0 1) ;双歧杆菌明显抑制了TNF α、IL 1β的表达 ;提高IL 10的表达。结论　双歧杆菌的作用可能与抑制NF κB的激活、降低TNF α、IL 1β等致炎因子的表达 ,提高IL 10等抑炎因子的表达有关