IL-1α、IL-1β和TNF-α mRNA 在实验性鼠根尖周炎中的表达

目的　检测IL-1α、IL-1β和TNF-αmRNA在鼠根尖周炎组织中的表达,了解上述细胞因子与根尖周炎的关系。方法　16只SD大鼠第一、二磨牙开髓,分别于术后3d、7d、14d、28d取磨牙根尖周组织,提取总RNA,用RT-PCR方法测定IL-1α、IL-1β和TNF-αmRNA的表达,用看家基因β-Actin作内参照进行半定量分析。结果　术后7d鼠根尖周组织中即有IL-1α、TNF-α、IL-1βmRNA的表达,14d达高峰,28d有所下降,但IL-1α和TNF-α的表达量大于IL-1β表达量,相差非常显著（P＜0.01）。结论　IL-1α和TNF-α可能是导致鼠根尖周炎的主要细胞因子。     

Expression of IL-6, IL-10, IL-17 and IL-8 in the peri-implant crevicular fluid of patients with peri-implantitis

Objectives

The aim of this study was to compare the levels of cytokines IL-6, IL-10 and IL-17 and the chemokine IL-8 in the peri-implant crevicular fluid (PCF) between the group of patients with peri-implantitis (PP) and peri-implantar healthy patients (HP).

Design

The PCF was collected from 40 implants regarding 25 patients, being 14 patients with PP and 11 HP totalizing 20 implants from each group. The PCF samples collected from each patient were quantified for IL-6, IL-17, IL-8 and IL-10 using the enzymatic immunosorbent assay (ELISA).

Results

The expression of IL-17 was significantly higher in the PP group when compared to HP (p < 0.05). There was no significant difference when comparing the levels of IL-6, IL-8 and IL-10 between both groups HP and PP. Also, there was a significant positive correlation between levels of IL-6 and IL-8 in the PP group.

Conclusions

This study demonstrates that in patients with peri-implantitis there is an increase of IL-17 which may induce the production of other inflammatory cytokines, contributing to the pathogenesis of bone loss in peri-implantitis.     

Expression of IL-6, IL-10, IL-17 and IL-33 in the peri-implant crevicular fluid of patients with peri-implant mucositis and peri-implantitis

ObjectivesThe aim of this study was to compare the levels of IL-6, IL-10, IL-17 and IL-33 in the peri-implantar crevicular fluid (PICF) and in parotid gland saliva (PGS) of healthy patients, and peri-implantitis and peri-implant mucositis patients.Materials and methodsThe PICF was collected from 40 implants as follows: 10 peri-implant mucositis patients, 20 peri-implantitis patients and 10 healthy patients. The PICF and PGS samples collected from each patient were quantified for IL-6, IL-10, IL-17 and IL-33 by enzymatic immunosorbent assay (ELISA).ResultsIL-6, IL-17 and IL-33 levels on PIFC were significantly higher in peri-implantitis group when compared to healthy group. IL-17 and IL-33 levels in PIFC were significantly higher in peri-implant mucositis group than in healthy group. There was no significant difference when comparing IL-6, IL-10, IL-17 and IL-33 levels in PGS among healthy, peri-implant mucositis and peri-implantitis groups.ConclusionsTherefore, as in patients with peri-implantitis there were significantly higher levels of IL-6, IL-17 and IL-33 in PICF, we believe that these cytokines were intensifying local inflammatory process, and contributing to clinical aspects such as increased marginal bleeding and probing depth found in patients with peri-implantitis. Furthermore, as IL-17 and IL-33 were increased in patients with peri-implant mucositis, hypothesized that these cytokines were also contributing to the inflammatory process observed in this disease.     

Gingival fluid IL-1 and IL-6 levels in refractory periodontitis

Abstract Selected gingival bacteria and cytokine profiles associated with patients who did not respond to conventional periodontal therapy (refractory) were evaluated. 10 subjects with a high incidence of post-active treatment clinical attachment loss (>2% sites/year lost ≥ 3 mm) were compared to 10 age-, race-, and supragingival plaque-matched patients with low post-treatment clinical attachment loss (<0.5% sites/year) relative to the following parameters at 2 sites/patient with the deepest probing depths: (1) presence of 3 selected periodontal pathogens (Actinobacillus antinomycetemcomitans, Porphyromonas gingivalis. Eikenella corrodens) in subgingival plaque as determined by selective culturing, and (2) gingival crevicular fluid (GCF) levels of 3 cytokines associated with bone resorption (IL-1 alpha, IL-1 beta, IL-6) as determined by two-site ELISA. Results indicated no significant differences in any clinical measurement (except incidence of clinical attachment loss), in the presence of any bacterial pathogen, or in GCF cytokine levels between refractory subject sites versus stable subject sites. However, when sites producing the greatest total GCF cytokine/patient were compared, sites from refractory patients produced significantly more IL-6 (30.1 ± 4.0 versus 15.4 ± 2.8 nM, p<0.01). The subgingival presence of each of the 3 bacterial pathogens was associated with elevated GCF IL-1 concentrations. These data suggest that gingival IL-1 and IL-6 production is different in response to local and systemic factors associated with periodontitis, and that IL-6 may play a role in the identification and mechanisms of refractory periodontitis.     

Force-induced IL-8 from periodontal ligament cells requires IL-1beta

During orthodontic tooth movement, mechanical stresses induce inflammatory reactions in the periodontal ligament (PDL). We hypothesized that chemokines released from PDL cells under mechanical stress regulate osteoclastogenesis, and investigated the profiles and mechanisms of chemokine expression by human PDL cells in response to mechanical stress. In vitro, shear stress and pressure force rapidly increased the gene and protein expressions of IL-8/CXCL8 by PDL cells. Consistently, amounts of IL-8 in the gingival crevicular fluid of healthy individuals increased within 2 to 4 days of orthodontic force application. The PDL cells constitutively expressed low levels of IL-1beta, which were not further increased by mechanical stress. Interestingly, neutralization of IL-1beta abolished IL-8 induction by mechanical stresses, indicating that IL-1beta is essential for IL-8 induction, presumably though autocrine or paracrine mechanisms. Finally, experiments with signal-specific inhibitors indicated that MAP kinase activation is essential for IL-8 induction.     

Simvastatin decreases IL-6 and IL-8 production in epithelial cells

Many cardiovascular studies have suggested that 3-hydroxy-3-methylglutaryl co-enzyme A reductase inhibitors (statins) have anti-inflammatory effects independent of cholesterol lowering. As a chronic inflammatory disease, periodontitis shares some mechanisms with atherosclerosis. Since oral epithelial cells participate importantly in periodontal inflammation, we measured simvastatin effects on interleukin-6 and interleukin-8 production by cultured human epithelial cell line (KB cells) in response to interleukin-1alpha. Simvastatin decreased production, an effect reversed by adding mevalonate or geranylgeranyl pyrophosphate, but not farnesyl pyrophosphate. Simvastatin was found to reduce NF-kappaB and AP-1 promoter activity in KB cells. Dominant-negative Rac1 severely inhibited interleukin-1alpha-induced NF-kappaB and AP-1 promoter activity. Our results may indicate an anti-inflammatory effect of simvastatin on human oral epithelial cells, apparently involving Rac1 GTPase inhibition.     

IL-1alpha, IL-1ra and IL-8 are differentially induced by Candida in experimental oral candidiasis

OBJECTIVE: To investigate the expression of interleukin-1alpha (IL-1alpha), IL-1ra and IL-8 by the oral epithelium challenged by various Candida species. MATERIALS AND METHODS: In vitro candidiasis was induced by C. albicans wild type SC5314, its EFG1, CPH1 and secretory aspartyl proteinase (SAP) mutants and, ATCC isolates of C. albicans, C. tropicalis and C. dubliniensis using a reconstituted human oral epithelium (RHOE) model. IL-1alpha, IL-1ra and IL-8 levels in culture media were quantified by an enzyme-linked immunosorbent assay at 12, 24 and 48 h. Fungal invasion and IL-1ra expression in RHOE were detected by periodic acid-Schiff staining and immunohistochemistry. RESULTS: Overall, the invasive Candida induced relatively higher levels of IL-1alpha, IL-1ra and IL-8 in the culture media than the noninvasive isolates. IL-1alpha and IL-1ra levels induced by Candida with hyphal invasion were significantly higher (P < 0.05) than those induced by the isolates without hyphal invasion at 12, 24 and 48 h. Candida albicans SC5314 induced IL-1ra expression in RHOE at 12 and 24 h but not at 48 h consistent with its hyphal invasion; while the noninvasive mutants and non-albicans Candida induced IL-1ra expression at 48 h. CONCLUSIONS: The cytokine expression profiles in experimental oral candidiasis may be associated with the invasive potential of Candida.     

IL-6、IL-33、IL-10在牙周炎和类风湿关节炎中的表达和相关性分析

目的: 检测牙周炎和类风湿关节炎患者龈沟液中白介素6（interleukin-6,IL-6）、白介素33（interleukin-33,IL-33）、白介素10（interleukin-10,IL-10）的含量,并探讨IL-6、IL-33、IL-10与牙周炎和类风湿关节炎2种疾病的相关关系。方法: 按纳入标准,选取就诊于中国医科大学附属盛京医院口腔科及风湿免疫科的患者,其中牙周炎组21例（PD组）,类风湿关节炎组21例（RA组）,类风湿关节炎合并牙周炎组24例（PD + RA组）,健康对照者24例（H组）。记录4组受试者一般信息、牙周探诊深度（periodontal probing depth,PPD）、临床附着丧失（clinical attachment loss,CAL）、龈沟出血指数（sulcus bleeding index, SBI）。收集各组受试者龈沟液样本,应用酶联免疫吸附试验测定IL-6、IL-33、IL-10的含量,并对4组受试者IL-6、IL-33、IL-10含量与牙周指标进行相关性分析。采用SPSS 20.0 软件包对数据进行统计学分析。结果: IL-6在PD+RA组的表达水平显著高于H组、PD组和RA组（P<0.05）; PD组、RA组、PD+RA组IL-33的含量显著高于H组（P<0.05）,PD+RA组IL-33的含量显著高于RA组(P<0.05);RA组IL-10的表达水平显著高于H组、PD组和PD+RA组(P<0.05)。PD组PPD与IL-6、IL-33的含量呈正相关（r=0.62, 0.43）,PD+RA组SBI、PPD、CAL与IL-33含量呈正相关（r=0.69,0.58,0.55）。结论: 牙周炎伴发类风湿关节炎患者龈沟液中IL-6、IL-33水平显著升高,IL-10含量显著降低,推测IL-6、IL-33和IL-10在牙周炎和类风湿关节炎的发生、发展中具有重要作用。     

IL-18与牙周病

IL-18是一种促炎症反应和肿瘤抑制因子[1],根据其结构、受体家族及信号转导途径,它属于IL-1家族细胞因子,可以诱导Th1和Th2的分化[2].尽管对其进行了一些研究,但究其与牙周病间的相关性研究还有待进一步探索.     

IL-4- and IL-6-producing cells in human periodontal disease tissue

IL-4- and IL-6-producing cells in human periodontal disease tissues were investigated using immunohistochemical and in situ hybridization techniques. Immunohistochemical analysis demonstrated the presence of IL-4-producing cells within the CD45RO⁺ subset and the percentage of IL-4+ cells was significantly higher in periodontal lesions than in gingivitis tissues (p<0.01). The percentage of IL-6-producing memory cells was higher in periodontal lesions compared with gingivitis tissues, although it was not statistically significant (p<0.05). A reverse tendency in IL-4- and IL-6-positive cells was observed in a few individual cases. No IL-4 mRNA could be detected using the in situ hybridization technique. However, high levels of IL-6 mRNA were present in clinically healthy tissues, with a further increase in both epithelium and connective tissues affected by gingivitis, although only the former was significant (p< 0.025). There was a significant decrease in IL-6 mRNA in both the connective tissue (p<0.025) and epithelium (p<0.01) in periodontitis tissues compared with levels in gingivitis tissues. However, the levels of IL-6 mRNA in periodontal tissues were high compared with those of IL-1 mRNA, which was used in this study as a positive control. These results suggest that Th2–type cells may accumulate in periodontal lesions.     

IL-1beta and TNF-alpha regulate IL-6-type cytokines in gingival fibroblasts

Interleukin-6 (IL-6)-type cytokines are pleiotropic molecules capable of stimulating bone resorption and expressed by numerous cell types. In the present study, we tested the hypothesis that gingival fibroblasts may exert local osteotropic effects through production of IL-6 and related cytokines. IL-6-type cytokine expression and regulation by IL-1beta and tumor necrosis factor-alpha (TNF-alpha) were studied in fibroblasts from the non-inflamed gingiva of healthy individuals. Constitutive mRNA expression of IL-6, IL-11, and leukemia inhibitory factor (LIF), but not of oncostatin M (OSM), was demonstrated, as was concentration-dependent stimulation of IL-6 and LIF mRNA and of protein by IL-1beta and TNF-alpha. IL-11 mRNA and protein were concentration-dependently stimulated by IL-1beta. The signaling pathway involved in IL-6 and LIF mRNA stimulation involved MAP kinases, but not NF-kappaB. The findings support the view that resident cells may influence the pathogenesis of periodontal disease through osteotropic IL-6-type cytokine production mediated by activation of MAP kinases. Abbreviations: IL-1alpha (interleukin-1alpha); IL-1beta (interleukin-1beta); IL-6 (interleukin-6); IL-11 (interleukin-11); LIF (leukemia inhibitory factor); OSM (oncostatin M); alpha(1)-coll. I (alpha(1)-collagen I); ALP (alkaline phosphatase); BMP-2 (bone morphogenetic protein-2); OC (osteocalcin); BSP (bone sialoprotein); TNFR I (tumor necrosis factor receptor I); TNFR II (tumor necrosis factor receptor II); IL-1R1 (interleukin-1 receptor 1); GAPDH (glyceraldehyde-3-phosphate dehydrogenase); RPL13A (ribosomal protein L13A); mRNA (messenger ribonucleic acid); cDNA (complementary deoxyribonucleic acid); PCR (polymerase chain-reaction); BCA (bicinchoninic acid); ELISA (enzyme-linked immunosorbent assay); alpha-MEM (alpha modification of Minimum Essential Medium); and FCS (fetal calf serum).     

Gingival fluid IL-1β and IL-6 levels in menopause

Abstract Menopause and oophorectomy without estrogen therapy (ED) have been associated with increased production of bone-active cytokines by peripheral blood mononuclear cells. The current study extended evaluation to gingival crevicular fluid (GCF) levels of interleukin (IL)-1β and IL–6 in such subjects compared to premenopausal and postmenopausal estrogen-treated females (ES). 13 ED and 13 ES Caucasians with a history of moderate-severe adult periodontitis provided GCF from 1–3 clinically identical sites each (5–6 mm probing depth, 5–7 mm clinical attachment loss, bleeding on probing). 30 s GCF samples were obtained and evaluated for IL-1β and IL-6 levels using two-site enzyme-linked immunosorbent assays (ELISAs). The frequency of GCF IL-1β-positive subjects was elevated in ED versus ES (92% versus 23%; p < 0.0004, X²; analysis). IL-6 was detected more frequently in ED subjects (23% versus 8%; not significant); however, the frequency of IL-6 detection was low in both groups due to short sampling times. These data support the concept that clinical conditions causing low estrogen environments allow increased local production of the bone-active cytokine IL-β, and perhaps IL-6.     

Investigation of functional gene polymorphisms IL-1beta, IL-6, IL-10 and TNF-alpha in individuals with recurrent aphthous stomatitis

Recurrent aphthous stomatitis (RAS) is characterized by recurrent episodes of oral ulceration in an otherwise healthy individual. Some reports in the literature indicate that RAS may have immunological, psychological, genetic and microbiological bases. OBJECTIVE: The purpose of the present study was to investigate, using binary logistic regression analyses, a possible association between the functional IL-1beta +3954 (C/T), IL-6 -174 (G/C), IL-10 -1082 (G/A) and TNF-alpha -308 (G/A) genetic polymorphism and RAS in a sample of Brazilian patients, using a multivariate statistical analysis. DESIGN: Sixty-four consecutive subjects affected by minor and major forms of RAS and 64 healthy volunteers were genotyped. To investigate the association between the single nucleotide polymorphisms and risk of RAS, binary logistic regression models were fitted. The associations were expressed by odd ratios (ORs) and adjusted for age and gender, with the corresponding 95% CIs. P-values less than 0.05 were considered significant. RESULTS: A significant increase in the IL-1beta and TNF-alpha heterozygous genotypes were associated with an increased risk of RAS development (OR 2.40 and 3.07, respectively), in the multivariate model. CONCLUSION: Our findings demonstrate that polymorphisms of high IL-1beta and TNF-alpha production were associated with an increased risk of RAS development. Our findings also give additional support to a genetic basis for RAS pathogenesis.     

IL-1 beta, IL-1 receptor antagonist and soluble type II IL-1 receptor in synovial fluid of patients with temporomandibular disorders

Among the members of the interleukin-1 (IL-1) family, IL-1 beta, which is a major agonist, has been detected in synovial fluid (SF) of the temporomandibular joint (TMJ) of patients with temporomandibular joint disorders (TMD). However, there is little knowledge regarding suppressive molecules, such as IL-1 receptor antagonist (IL-1ra) and the soluble form of type II IL-1 receptor (sIL-1RII), in TMD patients. The aim of this study was to investigate the levels of IL-1 beta, IL-1ra and sIL-1RII in the TMJ SF of patients with TMD and their relationship. Fifty-two SF samples from TMD patients and nine samples from asymptomatic volunteers were examined. Detected levels of IL-1 beta and sIL-1RII were significantly higher in the TMD group compared with the volunteer group. There was no significant difference in IL-1ra levels between the TMD and volunteer groups. The IL-1 beta/IL-1ra ratio in the TMD group, however, was higher than that in the volunteer group. In the TMD group, positive correlations were found between IL-1 beta and IL-1ra, IL-1ra and sIL-1RII, and IL-1 beta and sIL-1RII. In addition to increased IL-1 beta, development of TMD may also lead to decreased IL-1ra and increased sIL-1RII in response to increasing IL-1.     

实验性种植体周围炎龈沟液中IL-10和IL-17的分析

目的：分析实验性种植体周围炎不同时期龈沟液（PISF）中IL-10和IL-17的变化,探讨其在种植体周围炎发生发展过程中的作用。方法：采用丝线结扎法建立狗的实验性种植体周围炎模型,种植体随机分为实验组A（丝线持续结扎17w）和实验组B（结扎8w后去除丝线并进行洁治）,以口腔内另一侧不进行丝线结扎的种植体为对照。各组种植体分别在丝线结扎时（基点）和基点后1、4、8、9、13、17周时记录牙周袋深度（PPD）、菌斑指数（PI）和改良龈沟出血指数（MBI）,采集种植体周围龈沟液,检测PISF的量以及其中IL-10和IL-17的含量。分析不同时间点和不同组别种植体各项临床指标、PISF量以及其中IL-10和IL-17的差异。结果：与对照组相比,丝线结扎后种植体周围PPD、PI和MBI均出现改变。在临床指标出现改变前,PISF的量以及其中IL-10和IL-17的含量开始变化。PISF的量随时间增加,8周后保持比较稳定的水平,洁治后PISF量降低,13周后与对照组无差异。IL-10的含量随时间降低,8周后保持比较稳定的水平,洁治后开始升高。IL-17的含量随时间增加,洁治后降低。结论：IL-10和IL-17与种植体周围炎的发生发展有密切关系,其含量可在一定程度上反映种植体周围组织的炎症状况,IL-17可能作为早期诊断和预测种植体周围炎的指标。     

Single-nucleotide polymorphisms in the IL-4 and IL-13 promoter region in aggressive periodontitis

INTRODUCTION: IL-4 and IL-13 polymorphisms have been shown to influence the susceptibility to systemic diseases. In this study, possible associations between the IL-4 -590 C-->T, IL-4 -34 C-->T, IL-13 -1112 C-->T and IL-13 -1512 A-->C promoter polymorphisms were investigated in subjects with generalized aggressive periodontitis (AgP) compared with healthy individuals. MATERIAL AND METHODS: Fifty-eight patients with diagnosis of generalized AgP and 51 matched healthy controls participated in the study. Blood samples were collected and DNA isolated. Molecular analyses were performed by PCR-RFLP in a blind fashion. Genotype and allele frequencies among study groups were compared using Fisher's exact test (alpha value: 0.05). Pearson's chi(2) test was used for analysis of Hardy-Weinberg equilibrium. RESULTS: The frequency of the IL-4 -590 T/T and IL-4 -34 T/T genotypes differed significantly between groups (p=0.05, 0.02, respectively), although the allele frequencies were similar. There was a higher frequency of the IL-4 -590 T/T and IL-4 -34 T/T genotypes in patients with AgP compared with controls. The genotype and allele frequencies of the IL-13 polymorphisms did not differ between groups. CONCLUSIONS: This study demonstrated an association between the IL-4 -590 T/T and IL-4 -34 T/T genotypes and AgP. Further research is necessary to prove if there is an association of these polymorphisms with AgP, and if the polymorphisms have a functional effect.     

Prevalence of IL-1A and IL-1B polymorphisms in a Greek population

OBJECTIVES: Given the possible ethnic diversity in distribution of genetic variants, aim of the present study was to estimate the prevalence of the polymorphisms of IL-1A+4845 and IL-1B+3954 in a Greek population of unknown periodontal status and to compare this prevalence with one from a group of patients with chronic (adult) periodontitis. MATERIAL AND METHODS: 110 healthy subjects of unknown periodontal status and 45 patients with chronic periodontitis were genotyped, using a PCR-based method and primers described in the literature. The differences in genotype, allele frequencies, allele carriage rate and presence of positive composite genotype as described by Kornman et al. (1997) were analyzed using Fisher's exact test and calculating two-sided p-value, odds ratios and confidence intervals. RESULTS: No differences were observed in any of the parameters tested between the two groups. CONCLUSIONS: Given the high prevalences of these polymorphisms observed in the general population,findings of the present study do not support a possible predictive value of the presence of allele 2 at IL-1A+4845 and IL-1B+3954 or the positive composite genotype for presence or absence of periodontal disease,in a Greek population.