小剂量白细胞介素-18联合白细胞介素-10在早期Ⅱ型胶原诱导性关节炎中的作用研究

目的 研究不同细胞因子微环境下,白细胞介素(IL)-18对早期Ⅱ型胶原诱导性关节炎(CIA)的作用.方法 CIA小鼠,发病前给予每只重组小鼠腹腔注射IL-18(0.2 μg/d)/IL-10(0.1 μg/d),IL-18(0.2 μg/d)IL-4(0.1 μg/d),IL-18(0.2 μg/d)IL-12(0.1 μg/d),连用5 d,用关节炎评分评估关节炎病情的进展,用反转录聚合酶链反虚(RT-PCR)测定CIA小鼠髌骨及邻近滑膜组织的细胞因子表达水平;用组织学染色评价膝关节的软骨和骨破坏.采用Wilcoxon rank检验. 结果 IL-18/IL-4组关节炎评分轻度降低.IL-18/IL-10联合治疗有效抑制了C1A小鼠的关节炎症反应,免疫第38天治疗组关节炎评分(0.12±0.20)显著低于对照组(0.29±0.19,P<0.05),阻止了滑膜部位的炎细胞浸润,防止了软骨的破坏.滑膜和其附近软骨的Th1型细胞因子[IL-18(0.22±0.06),IL-12(0.14±0.05)]与炎症性细胞因子[IL-6(0.22±0.11)]水平与对照组相比显著降低(P<0.05),Th2型细胞因子[IL-10(6.35±0.12),IL-4(3.57±0.13)]水平与对照组相比明显升高(P<05). IL-18R(0.40±0.15)水平明显降低(P<0.05).T-bet水平降低,转录因子(GATA)-3水平(5.7l±0.11)显著升高(P<0.05).结论 IL-18/IL-10免疫干预尚未发病的CIA小鼠,可以抑制Th1型免疫反应,使免疫反应向Th2逆转,其机制可能为下调了IL-18/IL-18R水平,诱导了转录因子GATA-3的表达.     

小剂量白细胞介素-18对胶原诱导性关节炎作用的研究

目的 研究小剂量白细胞介素(IL)-18对早晚期Ⅱ型胶原诱导性关节炎(CIA)的作用.方法 重组小鼠IL-18(0.2 μg/小鼠)每日1次腹腔注射发病前或发病晚期的CIA小鼠,连用5 d,用关节炎评分评估关节炎病情的进展,用反转录聚合酶链反应(RT-PCR)测定CIA小鼠髌骨及邻近滑膜组织的细胞因子表达水平;用组织学染色评价膝关节的软骨和骨破坏.结果 小剂量IL-18早期免疫干预CIA小鼠,不能降低CIA小鼠关节炎发病率,无法延缓CIA小鼠膝关节病理学进展;滑膜和软骨附近的Th1型细胞因子和Th2型细胞因子与对照组相比差异无统计学意义.小剂量IL-18晚期免疫干预CIA小鼠,加剧了病理学进展,免疫第43天,治疗组关节炎评分(0.33±0.11)显著高于对照组(0.25±0.09)(P<0.05);髌骨及邻近滑膜组织的细胞因子IL-10与IL-18结合蛋白(IL-18BP)与对照组相比显著降低(P<0.05).结论 IL-18对早期Ⅱ型胶原诱导的关节炎没有疗效.IL-18晚期免疫干预CIA小鼠,恶化了关节炎病情,其机制可能为IL-18抑制了IL-10和IL-18BP的表达.     

2型糖尿病患者IL-10、IL-18与微量白蛋白尿的相关性

目的探讨2型糖尿病（T2DM）患者24h尿微量白蛋白排泄率（UAER）与IL-10、IL-18的相关性。方法检测正常对照组、单纯T2DM组（DM组）及伴微量白蛋白尿的T2DM组（DM-I-UAER组）的IL-10、IL-18水平,并分析其与UAER的关系。结果①DM组及DM＋UAER组的IL-18水平明显高于正常对照组,IL-10水平明显低于正常对照组（P均〈0．05）。②DM组与DM＋UAER组的IL-18水平比较有统计学差异（P〈0．05）,IL-10水平元统计学差异（P〉0．05）。③HbA,c、SBP、年龄、IL-18与UAER均呈显著正相关（P〈0．05）。结论IL-18与微量白蛋白尿的形成有关,因此可作为糖尿病肾病的早期预测指标;IL-10可作为T2DM的早期筛查指标。     

血清IL-10、IL-18与急性冠脉综合征的相关性研究

对45例急性冠脉综合征（ACS）患者（ACS组）、40例稳定型冠心病患者（SCHD组）及30例正常对照者的血清白细胞介素（IL-10）、IL-18水平进行测定,结果ACS组血清IL-10水平显著低于SCHD组和对照组（P均〈0．01）,血清IL-18水平显著高于SCHD组和对照组（P均〈0．01）;ACS患者血清IL-10与L-18呈显著负相关（P〈0．01）。提示血清IL-10、IL-18与不稳定粥样斑块破裂密切相关,两者共同作用促使ACS发生。     

亚健康状态老年人血白细胞介素18和白细胞介素10水平变化及意义

目的探讨白细胞介素18（IL-18）、白细胞介素10（IL-10）在老年亚健康状态发生发展中的作用及相互关系。方法应用酶联免疫吸附法测定45例亚健康状态老年人（≥60岁）血浆IL-18和血清IL-10水平,以42例亚健康状态老年前期（50～59岁）和30例健康老年人（≥60岁）作比较,分析它们之间的关系。结果亚健康状态老年人血浆IL-18水平显著高于健康老年人（P〈0.01）,以及亚健康老年前期组（P〈0.05）,血清IL-10水平则显著低于对照组（P〈0.01）和老年前期组（P〈0.05）;血浆IL-18水平与血清IL-10水平之间呈负相关（r=-0.882,P〈0.01）。结论 IL-18和IL-10可能共同构成一局部网络,并可能参与亚健康状态的发生发展过程。     

Polymorphisms in interleukin-10 gene according to mutations of NOD2/CARD15 gene and relation to phenotype in Spanish patients with Crohn＇s disease

AIM:To examine the contribution of interleukin-10(IL-10)gene polymorphisms to Crohn's disease(CD)phenotype,and the possible genetic epistasis betweenIL-10 gene polymorphisms and CARD15/NOD2 genemutations.METHODS:A cohort of 205 Spanish unrelated patientswith Crohn's disease recruited from a single centerwas studied.All patients were rigorously phenotypedand followed-up for at least 3 years(mean time,12.5years).The clinical phenotype was established prior togenotyping.RESULTS:The correlat on of genotype-Viennaclassification groups showed that the ileocolonic locationwas significantly associated with the-1082G allele in theNOD2/CARD15 mutation-positive patients(RR=1.52,95%CI,1.21 to 1.91,P=0.008).The multivariate analysisdemonstrated that the IL-10 G14 microsatellite allelein the NOD2/CARD15 mutation positive patients wasassociated with two risk factors,history of appendectomy(RR=2.15,95%CI=1.1-4.30,P=0.001)and smokinghabit at diagnosis(RR=1.29,95%CI=1.04-4.3,P=0.04).CONCLUSION:In Spanish population from Madrid,inCD patients carrying at least one NOD2/CARD15 mutation,the-1082G allele is associated with ileocolonic disease and the IL-10G14 microsatellite allele is associated withprevious history of appendectomy and smoking habit atdiagnosis.These data provide further molecular evidencefor a genetic basis of the clinical heterogeneity of CD.     

冠心病患者血浆组织因子、组织因子途径抑制物、白介素18、白介素10水平的变化及其关系

目的观察冠心病患者血浆组织因子(TF)、组织因子途径抑制物(TFPI)、白介素18(IL-18)、白介素10(IL-10)的变化,探讨其在冠心病(CHD)中的意义及其相互关系。方法 121例患者分为对照组(33例)与CHD组(96例),CHD组又分为急性心肌梗死(AMI)(33例),不稳定性心绞痛(UAP)(33例)和稳定性心绞痛(SAP)(30例)三个小组。采用酶联免疫吸附法分别测定血浆TF、TFPI、IL-18、IL-10的含量。结果 CHD组患者血浆TF、TFPI、TF/TFPI高于对照组,AMI小组患者血浆TF、TFPI、TF/TFP显著高于UAP及SAP小组患者,UAP小组患者高于SAP小组患者;CHD组患者血浆IL-18、IL-10、IL-18/IL-10高于对照组,AMI小组患者血浆IL-18、IL-18/IL-10显著高于UAP及SAP小组患者,UAP小组患者高于SAP小组患者;AMI和UAP小组患者血浆IL-10水平低于SAP小组患者;CHD组患者血浆TF与IL-18呈显著正相关(r=0.753,P=0.03),TF/TFPI与IL-18/IL-10呈正相关(r=0.496,P=0.01)。结论 CHD患者促凝因素和促炎症因子均显著升高,且与病情平行。促凝因子和促炎因子呈正相关,提示凝血因子和炎症因子在CHD发病中起重要作用,并可作为病情严重程度评价的参考指标。     

中药加味玉屏风颗粒对ACD小鼠IL-10和IL-18表达的影响

目的 探讨中药加味玉屏风颗粒对变应性接触性皮炎(ACD)小鼠IL-10和IL-18表达的影响.方法 建立小鼠ACD模型,分组给药,采用ELISA技术对各组外周血清IL-10和IL-18的表达水平进行检测.结果 与生理盐水组比较,加味玉屏风颗粒中高剂量组外周血IL-10、IL-18水平均明显下降(P<0.05).结论 中药加味玉屏风颗粒抑制小鼠ACD的作用机制可能与其下调机体的IL-10及IL-18相关.     

冠心病患者血浆IL-10,IL-17,IL-18和C反应蛋白的水平检测

目的:探讨血浆白细胞介素(IL)-10,IL-17,IL-18和C反应蛋白(CRP)在冠心病发病过程中的作用及其临床意义。方法:采用酶联免疫双抗体夹心法(ELISA)对160例冠心病患者(急性心肌梗死50例,不稳定型心绞痛68例,稳定型心绞痛42例)和40例正常人血浆中IL-10,IL-17,IL-18和CRP的水平进行检测。结果:冠心病患者血浆IL-10,IL-17,IL-18和CRP的水平均显著高于正常对照组(P<0.05)。结论:IL-10作为冠心病的保护因子,不足以阻止其发生、发展,IL-17,IL-18和CRP的水平与冠心病的发生与发展相关,其水平的检测对冠心病诊断及病情判断有重要意义。     

Analysis for expression and significance of interleukin-17, RORγt in CD4+ T cells from patients with ankylosing spondylitis

Objective To study the expression of IL-17, RORγt in CD4+T cells from patients with ankylosing spondylitis (AS). Methods The specimens of venous blood PBMC were collected from 28 patients with AS and 15 healthy subjects. Intracellular flow cytometry detection of IL-17 was established after isolation of human CD4+ T cells from PBMC. The expression level of IL-17, RORγt mRNA in CD4+ T cells was determined from 28 AS patients and 15 healthy controls by real-time fluorescence quantitative RT-PCR using Anti -CD3/Anti -CD28 as stimulators or not. Analysis of variance and Pearson correlation were selected. Results The isolation of human CD4+ T cells from PBMC was effective and its purity reached 90%. The percentage of intracellular IL-17 in CD4+ T cells from AS pati-ents in the AS active group was higher than that of the AS stable group and healthy control group (P<0.01). The expression level of IL-17, RORγt mRNA in CD4+ T cells was significantly higher in patients with AS than in controls. After stimulated with anti-CD3/ anti-CD28 stimulation, the percentage of IL-17, RORγt mRNA was increased significantly (P<0.01). The percentage of IL-17, RORγt mRNA in the 12 h group was higher than that of the 24 h group, while both of them were higher than those without stimulation (P<0.05). Conclusion There is an abnormal expression of IL-17, RORγt in human CD4+ T cells from AS patients. Our results indicate that the abnormal expression of IL-17 might play a role in the development and progression of AS.     

Analysis for expression and significance of interleukin-17, RORγt in CD4+ T cells from patients with ankylosing spondylitis

Objective To study the expression of IL-17, RORγt in CD4+T cells from patients with ankylosing spondylitis (AS). Methods The specimens of venous blood PBMC were collected from 28 patients with AS and 15 healthy subjects. Intracellular flow cytometry detection of IL-17 was established after isolation of human CD4+ T cells from PBMC. The expression level of IL-17, RORγt mRNA in CD4+ T cells was determined from 28 AS patients and 15 healthy controls by real-time fluorescence quantitative RT-PCR using Anti -CD3/Anti -CD28 as stimulators or not. Analysis of variance and Pearson correlation were selected. Results The isolation of human CD4+ T cells from PBMC was effective and its purity reached 90%. The percentage of intracellular IL-17 in CD4+ T cells from AS pati-ents in the AS active group was higher than that of the AS stable group and healthy control group (P<0.01). The expression level of IL-17, RORγt mRNA in CD4+ T cells was significantly higher in patients with AS than in controls. After stimulated with anti-CD3/ anti-CD28 stimulation, the percentage of IL-17, RORγt mRNA was increased significantly (P<0.01). The percentage of IL-17, RORγt mRNA in the 12 h group was higher than that of the 24 h group, while both of them were higher than those without stimulation (P<0.05). Conclusion There is an abnormal expression of IL-17, RORγt in human CD4+ T cells from AS patients. Our results indicate that the abnormal expression of IL-17 might play a role in the development and progression of AS.     

急性冠状动脉综合征患者血清白细胞介素-10、18及肿瘤坏死因子α水平及其临床意义

目的:评价血清白细胞介素(IL)-18、肿瘤坏死因子α(TNF-α)和IL-10水平与急性冠状动脉综合征(ACS)的关系及其临床意义。方法:冠心病(CHD)患者91例,分为ACS组、稳定型心绞痛(SAP)组;非CHD患者55例作为对照组,采用ELISA法测定血清IL-18、TNF-α和IL-10浓度并分析。结果:CHD患者血清IL-18和TNF-α水平明显高于对照组(P<0.01),而血清IL-10明显低于对照组(P<0.01);与SAP组相比,ACS组血清IL-18和TNF-α水平明显升高(P<0.01),而血清IL-10水平则显著降低(P<0.01)。且随着冠状动脉狭窄程度的增加,IL-18的水平逐渐增高。结论:IL-18、TNF-α和IL-10可以反映动脉粥样硬化斑块的严重程度和稳定状态,可作为监测病情的临床生化指标。     

葡萄籽原花青素对肾血管性高血压大鼠动态血压、白细胞介素-18和白细胞介素-10水平的影响

目的探讨葡萄籽原花青素(GSP)对肾血管性高血压(RH)大鼠动态血压、血清白细胞介素(IL)-18和IL-10水平和腹主动脉、肾皮质和海马组织中IL-10蛋白表达的影响。方法采用两肾一夹(2K1C)法,建立RH大鼠模型,并设立假手术组(n=7)。术后2 w,选取鼠尾动脉收缩压(SBP)超过130 mm Hg的大鼠28只为RH大鼠,并随机分为4组(n=7):模型组;低剂量GSP组(50 mg·kg-1·d-1);高剂量GSP组(200 mg·kg-1·d-1)及卡托普利组(30 mg·kg-1·d-1)。观察各组大鼠尾动脉SBP的动态变化,治疗6 w后,采用ELISA法检测各组大鼠血清中IL-18和IL-10含量,Western印迹法检测各组大鼠腹主动脉、肾皮质及海马组织中IL-10蛋白表达水平。结果与模型组相比,用GSP治疗2 w即可使RH大鼠尾动脉SBP降低(P0.05,P0.01),用药6 w后降压效果更明显(P0.05,P0.01)。同时亦能显著增高RH大鼠血清IL-10含量和腹主动脉、肾皮质及海马组织IL-10蛋白表达水平,降低RH大鼠血清IL-18含量,其中以高剂量GSP组作用尤为明显(P0.01),与卡托普利组的作用相当(P0.05)。结论 GSP可以部分通过增加RH大鼠体内抗炎因子IL-10的生成,减少促炎因子IL-18的产生而发挥抗炎和降压作用。     

白细胞介素-15、18和10与乙型肝炎相关性研究

乙型肝炎发病机制十分复杂,至今未完全阐明。各种细胞因子共同参与并相互影响,形成一个复杂而庞大的网络系统。本研究对血清白细胞介素（IL）-15、IL-18和IL-10水平与乙型肝炎相关性进行探讨。     

Serum interleukin-1 receptor antagonist is an early indicator of colitis onset in Gαi2-deficient mice

AIM: To study the serum concentration of IL-1beta, IL-1 receptor antagonist (IL-1Ra) and IL-18 in Galphai2-deficient mice at the age of 6 (healthy), 12 (pre-colitic) and 24 wk (colitic) and in healthy control mice. METHODS: At the time of killing, serum samples were collected and IL-1beta, IL-1Ra and IL-18 levels were measured using enzyme-linked immunosorbent assays. RESULTS: Serum concentration of IL-1Ra was significantly increased in pre-colitic (median: 524 ng/L; P=0.02) and colitic (450 ng/L; P=0.01), but not in healthy (196 ng/L) Galphai2-deficient mice as compared with controls (217 ng/L). Serum concentrations of IL-1beta did not differ between Galphai2-deficient mice and their controls, irrespective of age, IL-18 was significantly increased in colitic, but not in pre-colitic mice compared with controls (510 ng/L vs 190 ng/L; P=0.05). CONCLUSION: The increased serum concentrations of IL-18 and IL-1Ra in established diseases are suggested as markers of ongoing colitis. Interestingly, the significantly increased serum concentration of IL-1Ra in pre-colitic mice is found to be an early marker of disease progression.     

Oral and nasal administration of chicken type Ⅱ collagen suppresses adjuvant arthritis in rats with intestinal lesions induced by meloxicam

AIM: To investigate the curative effects of oral and nasal administration of chicken type Ⅱ collagen (CⅡ) on adjuvant arthritis (AA) in rats with meloxicam-induced intestinal lesions.METHODS: AA model in Sprague-Dawley (SD) rats with or without intestinal lesions induced by meloxicam was established and those rats were divided randomly into six groups which included AA model, AA model+meloxicam, AA model+oral CⅡ, AA model+nasal CⅡ, AA model+meloxicam+oral C Ⅱ and AA model+meloxicam+nasal CⅡ(n = 12). Rats was treated with meloxicam intragastrically for 7 d from d 14 after immunization with complete Freund‘s adjuvant (CFA), and then treated with chicken CⅡ intragastrically or nasally for 7 d. Histological changes of right hind knees were examined. Hind paw secondary swelling and intestinal lesions were evaluated. Synoviocyte proliferation was measured by 3-(4,5-dimethylthiazol-2-thiazolyl)-2,5-diphenyl-2H tetrazolium bromide (MTT) method. Activities of myeloperoxidase (MPO) and diamine oxidase (DAO) from supernatants of intestinal homogenates were assayed by spectrophotometric analysis.RESULTS: Intragastrical administration of meloxicam (1.5 mg/kg) induced multiple intestinal lesions in AA rats. There was a significant decrease of intestinal DAO activities in AA+meloxicam group (P&lt;0.01) and AA model group (P&lt;0.01) compared with normal group. DAO activities of intestinal homogenates in AA+meloxicam group were significantly less than those in AA rats (P&lt;0.01). There was a significant increase of intestinal MPO activities in AA+meloxicam group compared with normal control (P&lt;0.01). Oral or nasal administration of CⅡ (20 μg/kg) could suppress the secondary hind paw swelling(P&lt;0.05 for oral CⅡ; P&lt;0.01 for nasal CⅡ), synoviocyte proliferation (P&lt;0.01) and histopathological degradation in AA rats, but they had no significant effects on DAO and MPO changes. However, oral administration of CⅡ (20 μg/kg) showed the limited efficacy on arthritis in AA+meloxicam model and the curative effects of nasal CⅡ (20 μg/kg) were shown to be more efficient than that of oral CⅡ (20μg/kg) both in AA model and in AA+meloxicam model (P&lt;0.05). CONCLUSION: Oral administration of CⅡ shows the limited efficacy on arthritis in AA rats with intestinal lesions, and nasal administration of CⅡ is more efficient than oral administration of CⅡ to induce mucosal tolerance in AA rats.     

N-acetyl-L-cysteine combined with mesalamine in the treatment of ulcerative colitis： Randomized,placebo-controlled pilot study

AIM： To evaluate the effectiveness and safety of oral N-acetyl-L-cysteine （NAC） co-administration with mesalamine in ulcerative colitis （UC） patients.
METHODS： Thirty seven patients with mild to moderate UC were randomized to receive a four-wk course of oral mesalamine （2.4 g/d） plus N-acetyl-L-cysteine （0.8 g/d） （group A） or mesalamine plus placebo （group B）. Patients were monitored using the Modified Truelove-Witts Severity Index （MTWSI）. The primary endpoint was clinical remission （MTWSI ≤ 2） at 4 wk. Secondary endpoints were clinical response （defined as a reduction from baseline in the MTWSI of ≥ 2 points） and drug safety. The serum TNF-α, interleukin-6, interleukin-8 and MCP-1 were evaluated at baseline and at 4 wk of treatment. RESULTS： Analysis per-protocol criteria showed clinical remission rates of 63% and 50% after 4 wk treatment with mesalamine plus N-acetyl-L-cysteine （group A） and mesalamine plus placebo （group B） respectively （OR = 1.71; 95% CI： 0.46 to 6.36; P = 0.19; NNT = 7.7）. Analysis of variance （ANOVA） of data indicated a significant reduction of MTWSI in group A （P = 0.046） with respect to basal condition without significant changes in the group B （P = 0.735） during treatment. Clinical responses were 66% （group A） vs 44% （group B） after 4 wk of treatment （OR = 2.5; 95% CI： 0.64 to 9.65; P = 0.11; NNT = 4.5）. Clinical improvement in group A correlated with a decrease of IL-8 and MCP-1. Rates of adverse events did not differ significantly between both groups.
CONCLUSION： In group A （oral NAC combined with mesalamine） contrarily to group B （mesalamine alone）, the clinical improvement correlates with a decrease of chemokines such as MCP-1 and IL-8. NAC addition not produced any side effects.     

Selective decrease in colonic CD56^＋ T and CD161^＋ T cells in the inflamed mucosa of patients with ulcerative colitis

AIM： To investigate the role of local colonic mucosal NK receptor-positive T （NKR＋ T） cells in the regulation of intestinal inflammation, we analyzed the population and function of these cells in ulcerative colitis （UC）. METHODS： Colonic mucosal tissues were obtained from colonoscopic biopsies of the descending colon from 96 patients with UC （51 endoscopically uninflamed, 45 inflamed） and 18 normal controls. Endoscopic appearance and histologic score at the biopsied site were determined by MaLts＇ classification. A single cell suspension was prepared from each biopsy by collagenase digestion. Two NKR^＋ T cell subsets, CD56^＋ （CD56^＋CD3^＋） T cells and CD161＋ （CD161^＋CD3^＋） T cells, were detected by flow cytometric analysis. Intracellular cytokine analysis for anti-inflammatory cytokine interleukin-10 （IL-10） was performed by in vitro stimulation with phorbol-myristateacetate （PMA） and ionomycin. RESULTS： CD56^＋ T cells and CD161^＋ T cells are present in the normal human colon and account for 6.7% and 21.3% of all mononuclear cells, respectively. The populations of both CD56＋ T cells and CD161^＋ T cells were decreased significantly in the inflamed mucosa of UC. In contrast, the frequency of conventional T cells （CD56 CD3^＋ cells and CD161CD3^＋ cells） was similar among the patient and control groups. The populations of NKR^＋ T cells were correlated inversely with the severity of inflammation, which was classified according to the endoscopic and histologic Marts＇ criteria. Interestingly, approximately 4% of mucosal NKR＋ T cells expressing IL-10 were detected by in vitro stimulation with PMA and ionomycin.CONCLUSION： Selective reduction in the population of colonic mucosal NKR＋T cells may contribute to the development of intestinal inflammation in UC.