IL-34在慢性根尖周病损组织中的表达及临床意义

目的:比较慢性根尖周病损组织和健康牙周膜组织中白细胞介素34(Interleukin-34,IL-34)的表达水平,探讨IL-34在慢性根尖周炎发病中的作用.方法:收集25例诊断为慢性根尖周炎患牙的根尖周组织作为病例组,22例因正畸拔除的健康牙的牙周膜作为对照组,应用实时荧光定量PCR(real-time PCR,RT-PCR)检测IL-34mRNA的表达:应用免疫组织化学法检测IL-34蛋白的表达,采用图像分析软件检测病例组中IL-34蛋白的表达水平.采用SPSS13.0软件包对数据进行统计学分析.结果:IL-34mRNA在慢性根尖周炎中的表达水平(3.53±3.07)显著高于对照组(1.07±0.76),IL-34蛋白在慢性根尖周病损组织中阳性表达,主要定位于淋巴细胞、浆细胞及巨噬细胞中,且病例组中IL-34蛋白表达水平显著高于对照组(P＜0.01).结论:IL-34可能与慢性根尖周炎症密切相关.     

IL-10mRNA及其蛋白在慢性牙周炎牙龈组织中的表达

目的检测IL-10 mRNA及其蛋白在慢性牙周炎牙龈组织中的表达及其组织细胞来源.方法随机选择12例慢性牙周炎翻瓣术患者作为牙周炎组,10例龈切术患者作为牙龈炎组,6例阻生牙拔除术患者作为健康对照组.分别采用原位杂交和免疫组化检测技术,检测各组牙龈标本中IL-10的表达.每组IL-10 mRNA及蛋白两种水平间的比较采用秩和检验;各组间数据的两两比较采用单因素方差分析.结果IL-10 mRNA及其蛋白在牙周局部牙龈组织均有表达,表达细胞类型有淋巴细胞、浆细胞、巨噬细胞及成纤维细胞等.IL-10在mRNA水平及蛋白水平表达无显著差异(P＞0.05)(牙龈炎组P＜0.05).牙周炎组IL-10表达强度显著高于健康对照组和牙龈炎组(P＜0.01),牙周炎组IL-10 mRNA表达强度显著高于健康对照组(P＜0.01),但与牙龈炎组比较差异无显著性(P＞0.05).结论IL-10在牙周组织中存在局部分泌机制.     

IL-1β与IL-10mRNA在牙龈组织中的表达

目的:检测IL-1β与IL-10mRNA在正常牙龈及牙周炎患者牙龈组织中的表达,探讨二者与炎症的关系及内源性抗炎因子IL-10对致炎因子IL-1β是否有拮抗作用。方法:采集14例成人牙周炎患者炎症区牙龈组织,6例正常牙龈组织,利用逆转录-聚合酶链反应检测其中IL-1β与IL-10mRNA的表达及其强弱程度。结果:成人牙周炎组牙龈组织IL-1βmRNA的阳性表达率为92.86%,IL-10mRNA阳性表达率为71.43%,二者与正常对照组相比均有显著性差异;两者之间无明显相关性;两者与临床指标间亦无明显相关性。结论:致炎性和抗炎性细胞因子均可在牙龈组织中表达;机体自身IL-10水平不足以完全拮抗IL-1β活性;IL-10对致炎性细胞因子IL-1β的调控作用有待进一步研究。     

牙周炎和健康人牙周组织中细胞焦亡水平的比较研究

目的:研究慢性牙周炎和健康人的牙周组织中细胞焦亡水平是否存在差异。方法:分别纳入全身健康、无牙周病史、牙列完整的患者9例作为健康对照组，慢性牙周炎III～IV期患者9例作为牙周炎组。收集患者的牙周组织，免疫组化染色对牙龈组织中消皮素D(GSDMD)、半胱天冬酶-1(caspase-1)、NLRP3炎症小体、白细胞介素-1β(IL-1β)、白细胞介素-18(IL-18)的表达水平及分布进行定性分析；实时定量PCR检测牙龈组织和牙周膜组织中GSDMD、caspase-1、NLRP3、IL-1β及IL-18 mRNA的表达差异。结果:免疫组化染色显示牙周炎组的牙龈组织中GSDMD、caspase-1、NLRP3、IL-1β、IL-18的阳染色面积均大于健康组(P<0.05)。实时定量PCR检测显示，牙周炎组牙龈组织和牙周膜组织中GSDMD、caspase-1、NLRP3、IL-1β、IL-18 mRNA的表达量明显高于健康组(P<0.05)。结论:慢性牙周炎牙周组织的细胞焦亡水平高于健康人，提示细胞焦亡可能参与了牙周炎的发生发展。     

牙周炎患者牙龈组织中IL-21表达的研究

目的：研究不同类型牙周炎患者牙龈组织中IL-21基因的表达,探讨其在牙周炎发病中的作用。方法：选择慢性牙周炎患者12例,侵袭性牙周炎8例,健康对照组8例,采用实时定量PCR方法定量检测IL-21 mRNA在牙龈组织表达情况。结果：慢性牙周炎、侵袭性牙周炎、健康对照组牙龈组织中均有IL-21 mRNA的表达,慢性牙周炎组和侵袭性牙周炎组IL-21 mRNA相对表达量分别为0.000534±0.000504和0.00602±0.000137,显著高于健康对照组0.000161±0.000352,差异有统计学意义(P<0.05)。慢性牙周炎和侵袭性牙周炎牙龈组织IL-21mRNA表达没有显著性差异（P>0.05）。结论：IL-21在慢性牙周炎发病机制中可能发挥重要作用。     

IL-1βmRNA、TNF-αmRNA在成人牙周炎患者牙龈组织中表达的研究

目的:检测白细胞介素1B(IL-1B)mRNA、肿瘤坏死因子A( TNF-A)mRNA在成人牙周炎患者牙龈组织中表达, 探讨IL-1B和TNF-A与牙周炎致病机理的关系。方法:采用RT- PCR方法检测IL-1BmRNA、TNF-AmRNA在19例成人牙周炎患者炎症牙龈和9例牙周健康者牙龈组织中表达。结果:IL-1BmRNA表达强度在成人牙周炎病变牙龈 (0.819?01045)显著高于正常牙龈(0.306?0.087)(t=3.71,P<0.01)。TNF-AmRNA表达强度在成人牙周炎病变牙龈(0.696?0.098)显著高于正常牙龈(0)(t=7.09,P<0.01)。结论:IL-1B和TNF-A作为炎症和骨吸收介质可能在牙周炎的发生发展中起一定的作用。     

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目的    从转录水平探讨牙龈组织中白细胞介素-8（IL-8）与慢性牙周炎的关系。方法    选择2000年3—9月四川大学华西口腔医学院门诊就诊的全身无系统性疾病的患者32例，分为牙周健康组和慢性牙周炎组，各16例。采用原位杂交法检测32例患者牙龈组织中IL-8 mRNA表达，分析IL-8 mRNA的检出率及IL-8分泌细胞在牙龈组织中的分布。结果    慢性牙周炎组IL-8 mRNA检出率高于牙周健康组，IL-8阳性细胞分布于牙龈上皮和结缔组织中。结论    IL-8 mRNA不仅存在于牙周炎组织中，在正常组织中也有检出，IL-8 mRNA致炎的具体机制尚有待于进一步研究。     

慢性牙周炎患者IL-17mRNA的表达及意义

目的：观察TL-17mRNA基因在慢性牙刷炎和健康牙龈组织中的表达水平变化,探讨TL-17在慢性牙周炎发生发展巾的作用。方法：选择慢性牙周炎患者23例,正常对照组15例,记录才周临床指标,采用Real—time PCR方法定量检测TL-17mRNA在牙龈组织中的表达。结果：慢性牙周炎组TL-17mRNA相对表达晕（0．00147＋0．00055）显著高于正常牙龈组（O．00047±0．00019）（P〈0．01）,并且慢性牙周炎组TL-17mRNA表达水平与牙龈指数（r=0．58,P〈0．01）、牙周袋探诊深度（r=0．57,P〈0．01）、附着丧失水平（r=0．49,P〈0．05）呈正相关。结论：Th17相关细胞凶子IL—17呵能住慢性牙周炎发病机制中发挥致炎作用。     

牙周治疗前后龈沟液中白细胞介素-6水平的变化

目的：探讨牙周洁利治对患牙龈沟液中IL-6水平的影响。方法：选取12例成人牙周炎患者的重度牙周炎换牙12颗,采集治疗前患牙的龈沟液并记录相关的临床指标。然后对患牙进行龈上洁治和龈下刮治,两周后,再次采集患牙的龈沟液并记录相关的临床指标。采用双抗体夹心ELISA法对牙周炎患牙龈沟液中IL-6水平进行检测。比较牙周洁刮治前后龈沟液中IL-6水平的差异。结果：经过牙周洁刮治,患牙龈沟中IL-6的水平明显降低,同时患牙的牙周临床指标也有明显的改善。结论：牙周基础治疗在缓解牙周炎患牙局部炎症的同时,也对患牙局部的IL-6水平产生明显影响。     

慢性牙周炎病变部位白细胞介素-8表达的初步研究

目的探讨牙龈组织中自细胞介素-8(IL-8)与慢性牙周炎的关系。方法采用免疫组化法检测32例牙龈组织样本(16例健康者，16例慢性牙周炎患者)，分析IL-8的检出率及IL-8分泌细胞在牙龈组织中的分布。结果慢性牙周炎患者中的IL-8的检出率均高于健康者，IL-8阳性细胞分布于牙龈的上皮和结缔组织中。结论研究结果提示IL-8参与了牙周炎的病理过程。     

Shh蛋白与慢性牙周炎炎症程度的相关性研究

目的:探讨Sonic Hedgehog(Shh)信号通路中Shh蛋白与慢性牙周炎炎症程度的相关性。方法:选择健康对照20例(健康对照组),轻度牙周炎患者20例(轻度牙周炎组),中重度牙周炎患者20例(中重度牙周炎组),收集其龈沟液样本。采用ELISA法检测龈沟液中Shh蛋白、白细胞介素-6(IL-6)及白细胞介素-10(IL-10)的水平。结果:Shh蛋白、IL-6在慢性牙周炎组的水平均高于健康对照组(P<0.05),且中重度慢性牙周炎组高于轻度慢性牙周炎组(P<0.05);IL-10在轻度慢性牙周炎组的水平高于健康对照组及中重度慢性牙周炎组(P<0.05),且健康对照组高于中重度慢性牙周炎组(P<0.05);Shh蛋白、IL-6水平与出血指数(BI)、探诊深度(PD)均呈正相关(P<0.05)。结论:Shh信号通路可能参与了慢性牙周炎的炎症反应过程。     

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。     

慢性牙周炎患者IL-8和弹性蛋白酶水平的研究

目的：探讨IL-8、弹性蛋白酶在慢性牙周炎患者龈沟液中的表达。方法：采用ELISA检测慢性牙周炎患者30名和健康对照组20名龈沟液中IL-8的总量和浓度，采用底物法检测相同人群龈沟液中细胞外弹性蛋白酶、总弹性蛋白酶的浓度和总量。采用SPSS11．0软件分析其差异性及相关性。结果：慢性牙周炎患者龈沟液中的IL-8浓度明显低于健康对照组（P〈0．01），而IL-8总量略高于健康对照组但无显著性差异（P〉0．05）；IL-8和细胞外弹性蛋白酶的浓度和总量在重度牙周炎组明显高于轻、中度牙周炎组（P〈0．01）。结论：不同浓度的IL-8和弹性蛋白酶参与了慢性牙周炎的炎症过程，二者的协同作用将加重慢性牙周炎的病情。     

Expression of HMGB1 and HMGN2 in gingival tissues, GCF and PICF of periodontitis patients and peri-implantitis

Background and objective

High mobility group chromosomal protein B1 (HMGB1) and N2 (HMGN2), two members of high mobility group (HMG) family, play important role in inflammation. The purpose of this study was to investigate the expression of HMGB1 and HMGN2 in periodontistis.

Materials and methods

The expression of HMGB1 and HMGN2 mRNA in gingival tissues and gingival crevicular fluid (GCF) in chronic periodontitis (CP), generalised aggressive periodontitis (G-AgP) patients and healthy subjects was detected by real-time PCR. The protein level of HMGB1 and HMGN2 in peri-implant crevicular fluid (PICF), peri-implant crevicular fluid of peri-implantitis (PI-PICF) and normal patients was determined by Western blotting. Furthermore, IL-1β, IL-6, IL-8, TNF-α and HMGB1 levels in GCF, PI-PICF and healthy-PICF samples from different groups were determined by ELISA.

Results

HMGN2 expression was increased in inflamed gingival tissues and GCF from CP and G-ApG groups compared to control group. HMGB1 expression was the highest in the gingival tissues and GCF from CP patients and was accompanied by increased concentrations of IL-1β, IL-6, IL-8 proinflammaory cytokines.

Conclusion

To our knowledge, this is the first study reporting that the expression of HMGB1 and HMGN2 was increased in the gingival tissues and GCF in CP and G-AgP and the PICF in PICF. Our data suggest that HMGB1 may be a potential target for the therapy of periodontitis and PI.     

Porphyromonas gingivalis antigen preferentially stimulates T cells to express IL-17 but not receptor activator of NF-kappaB ligand in vitro

Although T cells have been implicated in the pathogenesis and are considered to be central to both their progression and control of chronic inflammatory periodontal diseases, the precise contribution of T cells to tissue destruction has not been fully clarified. Recently, interleukin (IL)-17 and receptor activator of Nuclear factor kappaB NF-kappaB ligand (RANKL) have received much attention as a result of their proinflammatory and bone metabolic roles, respectively. We therefore investigated the effect of outer membrane protein (OMP) from Porphyromonas gingivalis (P. gingivalis) on the expression of IL-17 and RANKL in peripheral blood mononuclear cells (PBMCs) and compared these between gingivitis and periodontitis, which are representative of stable and progressive lesions, respectively. The in situ expression of these molecules was also examined. P. gingivalis OMP stimulated PBMCs to express IL-17 at both the mRNA and protein level. Although the mean expression of mRNA was not different between the two groups, the mean level of IL-17 in the culture supernatants was higher in gingivitis patients than in periodontitis patients. However, the frequency of IL-17-positive samples was higher in the periodontitis patients. This stimulatory effect was not evident for RANKL expression in either periodontitis or gingivitis patients. In gingival tissue samples, IL-17 mRNA was detected in gingivitis more frequently than in periodontitis. The expression of RANKL mRNA was much lower than that of IL-17 in terms of both level and frequency. These results suggest that IL-17 but not RANKL may be involved in the pathogenesis of periodontal diseases. However, there may be negative regulatory mechanisms for IL-17 in gingivitis.     

DNA methylation status of the IL8 gene promoter in oral cells of smokers and non-smokers with chronic periodontitis

Aim: This study analysed the status of DNA methylation in the promoter region of the IL8 gene in oral mucosa cells from healthy, smoker and non-smoker subjects with chronic periodontitis and compared these findings among groups with mRNA levels.
Material and Methods: Genomic DNA from epithelial oral cells of 41 healthy subjects, 30 smokers with chronic periodontitis and 40 non-smokers with chronic periodontitis were purified and modified by sodium bisulphite. Genomic DNA from blood leucocytes and gingival cells from biopsies of 13 subjects of each group were also purified and modified by sodium bisulphite. Modified DNA was submitted by methylation-specific polymerase chain reaction (PCR) (MSP), electrophoresed on 10% polyacrylamide gels and stained with SYBR Gold. Total RNA from gingival cells was also isolated using the TRIzol reagent, and real-time PCR performance was used to detect the levels of interleukin-8 mRNA.
Results: Our results indicate that individuals with chronic periodontitis, independent of smoking habit, have a higher percentage of hipomethylation of the IL8 gene than those controls in epithelial oral cells ( p <0.0001), and expression of higher levels of interleukin-8 (IL-8) mRNA than controls in gingival cells ( p= 0.007). No significant differences among groups were observed in gingival cells and blood cells.
Conclusion: We conclude that inflammation in the oral mucosa might lead to changes in the DNA methylation status of the IL8 gene in epithelial oral cells.