IL—1家族与牙周炎研究的新进展

ＩＬ－１家族包括ＩＬ－１α，ＩＬ－１β和ＩＬ－１受体拮抗剂（ＩＬ－１ｒａ）。本文综述了ＩＬ－１家族各成员与牙周炎发生，发展关系的研究进展。并描述了ＩＬ－１ｒａ的临床前期动物模型研究成果，指出了ＩＬ－１ｒａ成为一种治疗牙周炎的非激素类抗炎新药有关的问题。     

白细胞介素—1与牙周膜的改建

白细胞介素－１（ＩＬ－１）是一种具有多种这活性的细胞因子，它亦是牙周莫成纤维细胞的重要调节因子，由于口腔这处特殊的环境，牙周膜总是承受着力的作用。本文就ＩＬ－１对牙周膜成纤维细胞的调节以及ＩＬ－１、力与牙周膜改建的关系及机理作一综述。     

内毒素诱导单核—巨噬细胞产生细胞因子的作用机理

内毒素刺激巨噬细胞产生，ＴＮＦ－α，ＩＬ－１，ＩＬ－６，ＩＬ－８，补体Ｃ３前列腺素（ＰＧ）等生物活性物质，在机体内发挥多种生物学，免疫学效应，细胞膜蛋白家族成人员之一的ＣＤ１４在内毒素诱导单核－巨噬分泌细胞因子的过程中起着重要作用，ＣＤ１４与ＬＰＳ－ＬＰＳ结合蛋白复合体（ＬＰＳ－ＬＢＰ）的结合发挥受体功能，抗ＣＤ１４抗体能够阻断ＬＰＳ－ＬＢＰ与单核细胞膜表面的结合，从而抑制细胞因子的产生。     

白细胞介素1β,肿瘤坏死因α对牙周组织的作用

目的：探讨白细胞介素１β（ｉｎｔｅｒｌｅｕｋｉｎ－１βＩＬ－１β）和肿瘤坏死因子α（ｔｕｍｏｒ ｎｅｃｒｓｏｓ ｆａｃｔｏｒ，α，ＴＮＦα）对牙周组织的作用。方法：将ＩＬ－１β，ＴＮＦα注入到兔的牙周组织，以生理盐水为对照，不同时期取材做组织学观察和评价。结果：实验组１５天后牙龈组织血管扩张充血，牙槽骨改建活跃，有大量的骨吸怍隐窝及破骨细胞，并有新骨形成和破骨细胞出现；     

破骨细胞活化因子及其在牙周炎发病中的作用

破骨细胞活化因子是一组具有诱导破骨细胞形成、刺激其活性、引起破骨吸收的生物活性因子，包括ＩＬ－１β，ＩＬ－１α，ＴＮＦα和ＴＮＦβ等。本文简述了这些细胞因子的产生及生物活性，重点讨论了这些细胞因子对骨吸收的调节作用，并就其与牙周病的关系作一论述。     

IL—1ra调节人牙周膜成纤维细胞ALP活性的实验研究

目的：观察人重组ＩＬ－１β对人牙周膜成纤维细胞（ＨｕｍａｎＰｅｒｉｏｄｏｎｔａｌＬｉｇａｍｅｎｔａｌｆｉｂｒｏｂｌａｓｔｓ，ＨＰＬＦ）的碱性磷酸酶（ａｌｋａｌｉｎｅｐｈｏｓｐｈａｔａｓｅ，ＡＬＰ）活性的影响以及ＩＬ－１ｒａ（Ｉｎｔｅｒｌｅｕｋｉｎ－１ｒｅｃｅｐｔｏｒａｎｔａｇｏ－ｎｉｓｔ，ＩＬ－１ｒａ）对ＩＬ－１β这一生物活性的阻断作用。方法：用碱性磷酸酶测定法。结果：经ＩＬ－１β作用后，ＨＰＬＦ的ＡＬＰ活性降低，经一定浓度的ＩＬ－１ｒａ作用后，ＩＬ－１β的这一作用相对减弱，ＡＬＰ活性增高。结论：ＩＬ－１ｒａ能够有效地阻断ＩＬ－１β对降低ＨＰＬＦ的ＡＬＰ活性的作用，进而有利于促进组织修复     

牙周炎患者龈沟液中IL—1活性检测

目的：通过对牙周炎龈沟液（ｇｉｎｇｉｖａｌｃｒｅｖｉｃｕｌａｒｆｌｕｉｄ，ＧＣＦ）中ＩＬ－１活性进行检测，初步探讨ＩＬ－１在牙周炎症中的作用。方法：以成人牙周炎为研究对象，通过细胞检测法对患牙ＧＣＦ中ＩＬ－１活性进行检测。结果：在患牙ＧＣＦ中可测到具有较高活性的ＩＬ－１，而在健康ＧＣＦ中未能测到。结论：牙周炎时局部有活性ＩＬ－１渗出，ＩＬ－１是参与牙周炎症反应的重要细胞因子。     

白细胞介素1受体拮抗剂的研究进展

白细胞介素－１受体拮抗剂是白细胞介素－１基因家族成员，它能与ＩＬ－１Ｒ结合。竞争性地阻断ＩＬ－１的活性。所以在ＩＬ－１介导的休克，炎症，白血症等疾病中有显著的疗效，对于研究ＩＬ－１病理生理作用和寻找新的临床生物调节剂具有重要意义。     

应用细胞因子单克隆抗体对人TMJ滑膜细胞的免疫组织化学研究

目的：了解人ＴＭＪ滑膜细胞正常状态下参与细胞因子合成能力及ＩＬ－１对这方面的影响和调控作用。方法：采用体外培养人ＴＭＪ滑膜细胞（ＳＭＣ）及４种细胞因子和抗ＩＬ－２ｒ抗体，以ＡＢＣ法，对正常状态下ＳＭＣ的培养细胞和经ＩＬ－１作用４ｄ后的细胞进行免疫组织化学染色，观察细胞浆、膜上阳性物质分布情况。结果：正常状态ＳＭＣ对ＩＬ－１、ＩＬ－２ｒ（白介素２受体）、ＩＬ－６、ＴＮＦ表达均呈阳性或弱阳性，主要分布在胞浆内，但经ＩＬ－１作用后，几种抗体染色的阳性细胞比例大幅增加，阳性颗粒密度也增加，整个视野内增加一倍以上。结论：正常状态下，ＳＭＣ就有少量细胞参加ＩＬ－１、ＩＬ－６、ＴＮＦ的合成，ＩＬ－１对这种合成能力有正相调节作用     

正常及炎症牙髓组织中白细胞介素1的活性检测

用细菌内毒素建立豚鼠的牙髓炎模型，借助体外培养的Ｌ９２９细胞，在培养液中加入正常和炎症的牙髓组织上清液，采用成纤维细胞增殖法测定正常和炎症牙髓组织中白细胞介素１（ＩＬ－１）的活性。结果发现正常牙髓组织无ＩＬ－１的活性，炎症牙髓组织产生明显的ＩＬ－１活性，并随机内毒素诱导时间的延长而增强，ＩＬ－１为牙髓炎的主要介质之一，介导了牙髓炎的发生和发展。     

Measurement of interleukin-1α and -1β in gingival crevicular fluid: Implications for the pathogenesis of periodontal disease

Samples of gingival crevicular fluid (GCF) were harvested from sites manifesting features characteristic of active disease including inflammation, periodontal attachment loss, and radiographic signs of alveolar bone destruction in untreated patients with advanced periodontitis. The presence and concentrations of interleukin-1 alpha (IL-1 alpha) and interleukin-1 beta (IL-1 beta) were measured using ELISAs specific for these cytokine molecules. IL-1 alpha and/or IL-1 beta were identified in the GCF of 15 of 15 patients having untreated periodontitis. Ninety percent (71 of 79) of the sites tested contained measureable amounts of IL-1, with IL-1 beta as the more frequently occurring form. IL-1 alpha levels ranged from 0.23 nM to 13.9 nM in the GCFs. IL-1 beta levels were between 0.04 nM and 5.28 nM. Marked reductions of total IL-1 levels were observed following effective treatment. Both forms of IL-1 messenger RNA (mRNA) were detected in 17 of 17 gingival tissue samples from 6 patients. These results demonstrate that IL-1 is produced and released locally in periodontal disease at concentrations sufficient to mediate tissue inflammation and bone resorption. IL-1 may serve as a marker of periodontal tissue destruction.     

Study of cytokine production in inflamed human gingival tissues in periodontitis. Interleukin-1 (IL-1 alpha, beta) and tumor necrosis factor (TNF alpha)

It seems to be generally agreed that periodontal disease is a local manifestation of a systemic immune response. Interleukin-1 (IL-1), which has multiple biologic activities, is detected in the gingival sulcus fluid of periodontitis sites. Recent investigations have revealed that IL-1 and tumor necrosis factor (TNF) are analogous to osteoclast activating factor and promote bone resorption. These findings have suggested the possibility that IL-1 and TNF may play a significant role in the initiation and development of periodontal disease. However, it remains to be determined whether these cytokines influence periodontal tissue breakdown in periodontitis. To elucidate the mechanisms of tissue breakdown in periodontitis, we examined cytokine production by human periodontitis gingival tissue. Twelve periodontitis patients were included in this study. Control subjects with healthy periodontium consisted of nine individuals. Gingival samples were biopsied from inflamed or healthy gingival tissues. Biopsy specimens were dissected into fragments 3 mm in diameter and plated onto 24 well culture plates with RPMI 1640 medium. IL-1 activity was measured by a growth inhibition assay using melanoma cell line A 375. An enzyme-linked immunosorbent assay (ELIZA) was used for measuring levels of human IL-1 alpha, IL-1 beta. TNF alpha activity was measured by a growth inhibition assay using cell line LM2D6. IL-1 activity was detected in significantly (p less than 0.001) higher levels in culture supernatants from gingival tissues in periodontitis (48.0 +/- 23.3 units/ml) than in control tissues (2.3 +/- 0.6 units/ml), however, levels of IL-1 activity were not associated with periodontal pocket depth or extent of alveolar bone resorption in periodontitis.(ABSTRACT TRUNCATED AT 250 WORDS)     

The influence of interleukin gene polymorphism on expression of interleukin-1beta and tumor necrosis factor-alpha in periodontal tissue and gingival crevicular fluid.

BACKGROUND: A specific composite genotype of the polymorphic interleukin-1 (IL-1) gene cluster has recently been associated with severe periodontitis. One polymorphism of the composite periodontitis-associated genotype (PAG) has been functionally linked with expression of high levels of IL-1. The purpose of this study was to test whether gingival crevicular fluid (GCF) levels of IL-1beta and tumor necrosis factor-alpha (TNFalpha), and gingival tissue levels of IL-1alpha, IL-1beta, and TNFalpha correlate with PAG, and to examine the effect of conservative periodontal therapy on these levels. METHODS: Twenty-two adults with moderate to advanced periodontal disease were enrolled. Polymerase chain reaction amplification and restriction enzymes were used to identify specific polymorphisms from peripheral blood samples. GCF samples were collected at baseline and 3 weeks following conservative treatment and analyzed by ELISA for IL-1beta and TNFalpha. An interproximal gingival biopsy was collected at baseline and follow-up and analyzed for IL-1alpha, IL-1beta, and TNFalpha by ELISA. RESULTS: The genotyping identified 7 as PAG(+) and 15 as PAG(-). The 2 groups were comparable in terms of existing periodontitis and age. In shallow sites (<4 mm), total IL-1beta in GCF was 2.5 times higher for PAG(+) patients prior to treatment (P=0.03), and 2.2 times higher after treatment (P=0.04), while differences were less apparent in deeper sites. Following treatment, a reduction in IL-1beta concentration in GCF was seen for PAG(-) but not for PAG(+) patients. While not statistically significant, a trend was observed in mean tissue levels of IL-1beta which were 3.6 times higher in PAG(+) versus PAG(-) patients (P=0.09). CONCLUSIONS: These data suggest that PAG(+) patients may demonstrate phenotypic differences as indicated by elevated levels of IL-1beta in GCF.     

Immunohistochemical evaluation of inflammatory mediators in failing implants

It was hypothesized that peri-implant tissue around loosening dental implants may contain cytokines with a potential to regulate osteoclasts. Peri-implant and/or gingival samples from loosened implants, chronic periodontitis (CP), and normal controls (n = 10 samples in each group) were analyzed using immunohistochemical staining to observe tumor necrosis factor alpha (TNF-alpha), interleukin 1-alpha (IL-1alpha), IL-6, platelet-derived growth factor A (PDGF-A), and transforming growth factor alpha (TGF-alpha). These cytokines were found in foreign-body giant cells, macrophages, fibroblasts, and epithelial cells. TNF-alpha, IL-1alpha, and IL-6 were increased (P < .05; unpaired t test) in peri-implantitis and CP, whereas PDGF-A and TGF-alpha were not. In conclusion, cytokines with a potential to activate osteoclasts were found in both peri-implantitis and CP, but the cytokine profiles differed in that IL-1alpha was the most prevalent cytokine in the former and TNF-alpha was the most common in the latter. These cytokines may contribute to peri-implant bone loss/loosening by stimulating formation and activity of osteoclasts and might be an amenable target for local therapies with cytokine modulators.     

牙周炎患者自身组织核酸内NLRP3 mRNA表达水平测定

目的:通过对牙周炎患者自身组织核酸内炎性小体复合物NLRP3,NLRC4及其相关基因IL-18,IL-1β,Caspase-1等mRNA表达水平的检测,明确炎性小体NLRP3在牙周炎中具有调控作用。方法:采集翻瓣术中慢性牙周炎患者炎性牙周组织以及正畸患者健康牙拔除术获取的健康牙周组织,提取总RNA逆转录cDNA ,-20 ℃冻存备用。Real-time PCR检测NLRP3、NLRC4、IL-18、IL-1β、Caspase-1 mRNA的表达。结果:与健康组牙周组织相比较,牙周炎患者自身组织核酸内NLRP3、IL-18、IL-1β、Caspase-1 mRNA的表达水平明显增高;而NLRC4 mRNA的表达在两组间差异不显著。结论:NLRP3及其相关基因IL-18、IL-1β、Caspase-1 mRNA在牙周炎患者自身组织核酸内的高表达,提示NLRP3在牙周炎的发病机制中具有影响作用。     

Relationship between IL-1A polymorphisms and gingival overgrowth in renal transplant recipients receiving Cyclosporin A

AIM: Levels of interleukin-1alpha (IL-1alpha) are elevated in periodontal inflammation. IL-1A gene polymorphisms are associated with inflammatory diseases. This study aimed to investigate IL-1A gene polymorphism in Cyclosporin A (CsA)-treated renal transplant patients and investigate the association between this polymorphism and gingival crevicular fluid (GCF) levels of several cytokines. MATERIALS AND METHODS: Fifty-one renal transplant patients on CsA treatment (25 with and 26 without gingival overgrowth) and 29 healthy controls were recruited for the study. Demographic, pharmacological and periodontal parameters were recorded and gingival overgrowth was assessed. RESULTS: Multiple regression analysis showed that genotype was significantly associated with gingival overgrowth (p=0.02). Carriage of the IL-1A (-889) T allele was strongly protective [95% confidence interval (CI): 0.046-0.77], although not significantly associated with IL-1alpha protein levels in GCF. IL-1alpha, IL-1beta and IL-8, but not IL-6, were detected in GCF of CsA-treated patients, but none of them was significantly associated with gingival overgrowth. CONCLUSIONS: This study is the first to associate a gene polymorphism as a risk factor for CsA-induced gingival overgrowth in renal transplant patients, demonstrating that IL-1A polymorphism might alter individual susceptibility to CsA. However, there was no association between GCF cytokine levels and the presence of gingival overgrowth or patient IL-1A genotype.     

The effect of IL-1alpha and nifedipine on cell proliferation and DNA synthesis in cultured human gingival fibroblasts

The effect of nifedipine and interleukin-alpha (IL-1alpha) on the cell proliferation and DNA synthesis was studied in human gingival fibroblasts derived from 5 patients who developed gingival overgrowth (nifedipine responders) and 5 patients who did not develop gingival overgrowth (nifedipine non-responders) in response to nifedipine. Epidermal growth factor was used as a positive control. The fibroblasts derived from nifedipine responders tended to have a numerically greater rate of cell proliferation and DNA synthesis (3H-thymidine incorporation) than those from nifedipine nonresponders in the presence of nifedipine and IL-lalpha. Fibroblasts derived from nifedipine responders showed significantly higher cell proliferation rate in the presence of nifedipine and IL-1alpha, than nifedipine or IL-lalpha alone on both the second and the fourth day of incubation (P < 0.05). A combination of IL-1alpha and epidermal growth factor also showed significantly greater cell proliferation than IL-lalpha alone on the second day (P < 0.05). The DNA synthesis rate with a combination of nifedipine and IL-1alpha was higher than that for nifedipine alone on the second day (P < 0.01), and IL-1alpha alone on the fourth day (P < 0.05) in gingival fibroblasts originating from nifedipine responders. These results suggest that the interaction between nifedipine and gingival inflammation might play an important role in the pathogenesis of nifedipine-induced gingival overgrowth.