内质网应激诱导的细胞凋亡在慢性牙周炎中的作用初探

利用光学显微镜和透射电镜观察牙周组织中的凋亡细胞,免疫组化染色法检测慢性牙周炎患者及牙周健康者牙周组织中GRP78和CHOP的分布及含量变化。结果:慢性牙周炎组内浆细胞呈现凋亡状态,且GRP78和CHOP在慢性牙周炎组中的阳性表达明显高于正常组(P<0.05)。结论:     

内质网应激诱导的细胞凋亡在慢性牙周炎中的作用初探

目的:初步探讨内质网应激诱导的细胞凋亡在慢性牙周炎中的作用。方法:利用光学显微镜和透射电镜观察牙周组织中的凋亡细胞,免疫组化染色法检测慢性牙周炎患者及牙周健康者牙周组织中GRP78和CHOP的分布及含量变化。结果:慢性牙周炎组内浆细胞呈现凋亡状态,且GRP78和CHOP在慢性牙周炎组中的阳性表达明显高于正常组(P<0.05)。结论:内质网应激参与了慢性牙周炎的病理生理过程。     

实验性糖尿病牙周炎诱导骨细胞凋亡的初步研究

目的初步探讨糖尿病牙周炎条件下骨细胞的凋亡情况。方法选用6wk雄性SD大鼠62只,随机分为糖尿病牙周炎组（DP,n=22）、牙周炎（P,n=20）以及正常对照组（N,n=20）。采用一次性腹腔注射STZ（55mg/kg）的方法建立大鼠糖尿病模型,注射STZ后1wk检测血糖,血糖≥16.65mmol/L者定为糖尿病大鼠。采用丝线结扎大鼠上颌第二磨牙联合口内接种细菌的方法建立牙周炎模型。动物分别于丝线结扎后3wk和6wk分批处死,进行HE染色和原位细胞凋亡检测。观察指标包括：牙槽骨丧失（ABL）,骨细胞计数,骨细胞凋亡百分率。资料采用单因素方差分析统计学处理。结果丝线结扎后3周和6周,大鼠牙槽骨丧失在N组与P组、N组与DP组、P组与DP组不同,组间两两比较均有统计学意义（P〈0.05）,牙槽骨丧DP组〉P组〉N组。与N组比较,P组和DP组单位面积骨细胞数均减少,与P组比较,DP组单位面积骨细胞数亦显著减少（P〈0.05）。在丝线结扎后3周和6周,糖尿病牙周炎组（DP）骨细胞凋亡百分率均达到牙周炎组（P）的2倍左右。结论糖尿病条件下牙周炎骨丧失明显增加,糖尿病可加强牙周炎条件下牙周组织中骨细胞的凋亡,降低骨细胞的数量。     

糖尿病对大鼠牙周组织及诱导型一氧化氮合酶分布的影响

目的：观察糖尿病对大鼠牙周炎牙周组织形态结构的影响及诱导型一氧化氮合酶（iNOS）在牙龈组织中的分布。方法：建立大鼠糖尿病牙周炎模型,制作组织切片,进行HE及免疫组织化学染色,观察牙周组织形态结构的破坏情况及iNOS在牙龈组织中的分布。结果：糖尿病牙周炎组的结缔组织附着丧失量及牙槽骨高度丧失量均明显高于牙周炎组、糖尿病组及正常组。差异有显著性（P〈0．05）。糖尿病牙周炎组和糖尿病组的免疫组化染色均为阳性或强阳性。结论：糖尿病加重了牙周炎牙周组织的破坏程度。     

2型糖尿病对大鼠牙周组织及TNF-α、IL-6和PAI-1水平影响的研究

目的观察2型糖尿病伴牙周炎大鼠牙周组织及血清肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)及纤溶酶原活化物抑制因子-1(PAI-1)水平的变化,探讨TNF-α、IL-6及PAI-1的表达对2型糖尿病牙周炎大鼠牙周组织改建过程中组织变化的影响。方法清洁级8~9周龄Wistar大鼠60只,分为4组:(1)伴牙周炎糖尿病组(n=15);(2)不伴牙周炎糖尿病组(n=15);(3)伴牙周炎非糖尿病组(n=15);(4)不伴牙周炎非糖尿病组(正常对照组,n=15)。采用腹腔内注射链脲佐菌素(streptozotocin,STZ)法建立大鼠糖尿病模型。给药后每周1次,连续4周测定大鼠尾静脉血糖水平,以判断大鼠糖尿病建模情况。用浸有牙龈卟啉单胞菌株的丝线结扎左侧上颌第二磨牙牙颈部,复制实验性牙周炎模型。实验性牙周炎建模20周后,采用腹腔注射水合氯醛法在全麻状态下处死大鼠。从腹主动脉采集血液,ELISA法检测血清TNF-α、IL-6及PAI-1水平。处死前测量术区的牙周附着情况。制作牙体牙周联合切片,观察牙周组织学改变。结果糖尿病组大鼠表现为血糖和体质量水平显著升高(P<0.05);糖尿病组大鼠血清胰岛素水平和胰岛素抵抗指数得分均显著升高(P<0.05);用浸有牙龈卟啉单胞菌株的丝线结扎后,牙周炎组表现为典型的牙周炎临床特征;与不伴牙周炎糖尿病组相比,伴牙周炎糖尿病组大鼠血清TNF-α、IL-6及PAI-1水平显著升高(P<0.05)。结论糖尿病通过促进血清炎性因子的表达,降低牙周组织对局部致病因子的抵抗力,从而加重、加速牙周炎的进展。     

不同牙周状态正畸力诱导破骨细胞分化因子RANK在牙周组织的表达

目的 研究不同牙周状态正畸力作用下破骨细胞分化因子RANK在牙周组织的表达变化,为牙周病正畸治疗提供参考.方法 建立大鼠牙周炎静止期、牙周炎活动期模型,以50克力移动牙周正常组、牙周炎静止期组、牙周炎活动期组的上颌磨牙.分别于牙齿移动的第3和第7天处死大鼠.采用免疫组化和实时荧光定量PCR定性定量分析各组RANK在牙周组织的表达.结果 正常牙周移动组RANKmRNA的表达高于正常对照组(P＜0.05).静止期牙齿移动组RANKmRNA在牙周组织的表达高于正常牙周移动组(P＜0.01).较活动期移动组显著减小(P＜0.01).牙周炎活动期牙齿移动组RANKmRNA的表达均高于其他各组(P＜0.01).结论 RANK参与调节正常牙周及牙周炎正畸牙齿移动.牙周炎静止期正畸力可诱导RANK的表达增加,但显著小于炎症活动期.此时要密切控制牙周易感因素.     

Etanercept作用下糖尿病Wistar大鼠牙周炎变化的实验研究

[摘要] 目的 观察肿瘤坏死因子α抑制剂Etanercept对实验性糖尿病Wistar大鼠牙周炎的影响。方法 12周龄Wistar大鼠45只,腹腔内一次性注射STZ诱导糖尿病。结扎糖尿病大鼠牙颈部建立牙周炎模型。将实验糖尿病大鼠分为糖尿病组,糖尿病牙周炎组和糖尿病牙周炎+Etanercept组。观察大鼠的牙龈指数、牙周袋深度、松动度。免疫组化观察IL-1β、IL-6在牙周组织中的表达。结果 糖尿病组大鼠牙龈指数、牙周袋深度、松动度与糖尿病牙周炎组和糖尿病牙周炎+Etanercept组有差别(P<0.05);糖尿病牙周炎组和糖尿病牙周炎+Etanercept组相比有差别(P<0.05)。糖尿病牙周炎组中IL-1β、IL-6表达高于糖尿病组(P<0.05),各时间段糖尿病牙周炎+Etanercept组牙周组织中IL-1β、IL-6表达低于糖尿病牙周炎组(P<0.05)。结论 肿瘤坏死因子α抑制剂Etanercept对实验性糖尿病Wistar大鼠牙周炎有抑制作用。     

釉基质衍生物对牙周炎症组织中凋亡发生的影响

目的：探讨釉基质衍生物（EMD）对牙龈卟啉单胞菌脂多糖（Pg—LPS）龈沟内注射形成的牙周炎症组织中细胞凋亡的影响。方法：隔日向SpragueDawley大鼠双侧上颌第一及第二磨牙间腭侧龈沟内注射Pg—LPS2wk建立牙周炎模型;牙周炎形成后,龈沟内注射EMD或磷酸盐缓冲液;截取上颌骨标本,行HE染色观察牙周组织破坏情况,TUNEL染色观察牙周组织中细胞凋亡,免疫组织化学染色观察凋亡酶caspase--3和波形蛋白的表达。结果：在大鼠龈沟内注射Pg—LPS后,牙周组织形成明显的炎症和附着丧失;TUNEL染色显示EMD可以抑制Pg—LPS所致牙周组织中的细胞凋亡;免疫组织化学染色显示EMD降低凋亡酶caspase--3的表达,并且促进牙周组织中波形蛋白的表达。结论：EMD可以抑制Pg—LPS所致牙周组织中的细胞凋亡,从而促进牙周组织再生。     

实验性糖尿病牙周炎诱导成骨细胞凋亡的研究

目的观察糖尿病牙周炎条件下成骨细胞的凋亡情况。方法选用SD大鼠62只,随机分为糖尿病牙周炎组（DP）、单纯牙周炎组（P）以及空白对照组（N）。采用一次性腹腔注射链脲佐菌素（STZ）的方法诱导大鼠糖尿病模型,采用丝线结扎联合口内接种细菌的方法建立牙周炎模型。各组动物分别于DP组、P组丝线结扎后第3、6周时分批处死,取标本进行组织学检测,并计算各组成骨细胞凋亡百分率。结果丝线结扎后3、6周时,成骨细胞凋亡百分率由高至低依次为DP组、P组、N组,组间两两比较均有统计学意义（P＜0.05）。在丝线结扎后3周和6周时,DP组成骨细胞凋亡百分率均达到P组的2倍。结论糖尿病可促进牙周炎条件下牙周组织中成骨细胞的凋亡。     

盐酸小檗碱对大鼠牙周组织中骨钙素、白细胞介素-6表达的影响

目的：探讨盐酸小檗碱对大鼠实验性牙周炎模型的牙周组织中骨钙素（bone gla protein,BGP）和白细胞介素-6（interleukin-6,IL-6）表达的影响。方法：在成功建立大鼠实验性牙周炎模型的基础上,实验动物分别灌服盐酸小檗碱后第1、2、3、4周末处死,取牙周组织标本作切片,常规HE染色,观察牙周组织病理变化状况,并采用免疫组化SABC法检测BGP、IL-6水平变化。结果：牙周炎组牙周组织中BGP表达明显低于正常组,IL-6表达明显高于正常组（P〈0.05）;治疗组BGP表达明显高于牙周炎组,IL-6表达明显低于牙周炎组（P〈0.05）。结论：盐酸小檗碱能促进大鼠实验性牙周炎模型牙周组织中BGP的表达,同时抑制IL-6的表达。     

Inhibition of SFRP1 reduces severity of periodontitis

Porphyromonas gingivalis (P. gingivalis) is implicated as a major pathogen in periodontitis, a common infectious disease characterized by the inflammation and destruction of periodontal tissues. Secreted frizzled-related protein 1 (SFRP1) modulates apoptosis in different cell types. To characterize the roles of SFRP1 in periodontitis, we used a P. gingivalis-induced murine periodontitis model. Inflammatory responses were measured by morphometric and histomorphometric analysis, apoptosis assay, and immunohistochemistry. We found that P. gingivalis-infected mouse periodontal tissues expressed significantly more SFRP1 compared with those of control mice. Also, in P. gingivalis-infected animals, more apoptosis of inflammatory cells, fibroblasts, and bone-lining cells was observed compared with controls. Antibody experiments aimed at inhibiting SFRP1 expression in periodontitis resulted in a reduction of periodontal breakdown, inflammatory cell infiltrate, osteoclastogenesis, and apoptosis of inflammatory cells and fibroblasts. The results of our studies suggest that SFRP1 may be involved in the development of periodontitis, since inhibiting SFRP1 resulted in reduced periodontal breakdown.     

Apoptosis: an underlying factor for accelerated periodontal disease associated with diabetes in rats

Objectives

Diabetes mellitus (DM) is well-established risk factor for periodontal disease. DM can also lead to changes in the number of apoptotic cells in periodontal tissues. The goal of this study was to evaluate apoptosis, depending on DM, in healthy and diseased periodontal soft tissues.

Material and Methods

A total of 43 adult male Sprague–Dawley rats were used in this study. Experimental periodontitis was created by placing silk ligatures around the cervices of the first mandibular molars. Experimental diabetes was induced by intraperitoneal injection of the diabetogenic agent streptozotocin (STZ). Following the induction of both experimental diseases, the animals were divided into four groups: (1) The healthy group (H) (n?=?10); (2) The diabetes group (D) (n?=?10); (3) The periodontitis group (P) (n?=?11); and (4) The diabetes and periodontitis group (DP) (n?=?12). Apoptotic cells were determined by immunohistochemistry, and the frequency of apoptotic cells was evaluated by apoptotic index score.

Results

It was observed that there was less apoptosis in both the epithelial and gingival connective tissue cells of healthy diabetic tissues than in healthy tissues without diabetes. When periodontal disease existed, apoptosis increased in both the epithelial and gingival connective tissues of diabetic and non-diabetic animals.

Conclusions

There may be differences in the apoptotic mechanisms in the periodontal soft tissues of diabetic and non-diabetic animals.

Clinical relevance

Apoptosis may be one of the underlying factors in increased risk for periodontal disease that is associated with diabetes.     

Periodontal disease in juvenile and adult diabetic patients: a review of the literature

Epidemiologic studies have suggested that the severity of periodontitis is greater in juvenile and adult onset diabetes. In juvenile diabetic patients, the periodontal disease seems to be initiated around puberty and progresses by age. Reviewing the medical literature indicates a similar age of onset for known systemic complications resulting from diabetes. Angiopathy, abnormal collagen metabolism, abnormal PMN function, and altered sulcular microbial flora have been found to be closely associated with the severity of periodontitis in diabetic patients. The association between abnormal neutrophil function and severity of periodontal disease in diabetic patients provides an opportunity for examining the role of neutrophil in periodontal disease. Future investigation in the function of sulcular PMN may shed light on the complex mechanism of periodontal disease.     

Diabetes mellitus and periodontal disease in an Irish population

The prevalence of periodontitis was studied in a population of 157 insulin dependent diabetes mellitus patients aged 8-78 years attending the outpatients diabetic clinic of a large general hospital in Cork, Ireland. Every third diabetic patient attending the clinic was selected for examination. The dental parameters measured were plaque index (PI), gingivitis index (GI), periodontal pocket depth (PD) and periodontal attachment loss (PAL). Diabetic control was measured by estimating percentage haemoglobin glycolysation (% Hb Alc) known duration of diabetes (KDD) and insulin dependence. It was found that none of the diabetic measurements showed any consistent pattern in relation to any of the periodontal measurements. The findings are in agreement with other studies which suggest that no significant correlation between diabetic parameters and periodontal disease can be demonstrated. When the diabetic patient suffered periodontitis it was due to factors (such as genetic predisposition) other than impaired glucose metabolism.     

Tumor necrosis factor-alpha expression and detection of apoptosis at the site of chronic periodontitis in AIDS patients

BACKGROUND: Apoptosis, or programmed cell death, is associated with the regulation of the life cell cycle of leukocytes in healthy and diseased states. OBJECTIVES: In the present study, we investigated the presence of apoptosis of mononuclear inflammatory cells in the periodontal lesion from adult periodontitis in healthy control patients and AIDS patients. MATERIALS AND METHODS: Tissue samples adjacent to a 5-6 mm gingival sulcus, measured with a periodontal probe, were obtained during routine periodontal surgical procedures. The direct immuno-peroxidase of digoxigenin-labeled genomic DNA method was used for in situ detection of apoptosis in gingival tissues. RESULTS: Many tumor necrosis factor-alpha (TNFalpha)-positive cells, detected by immunohistochemistry method, were observed in gingival samples of both groups of patients. In addition, a significant lower number ( p < 0.05) of mononuclear apoptotic cells were observed in AIDS patients when compared with healthy control patients. CONCLUSION: These data suggested an important role of the apoptosis of mononuclear cells in the pathogenesis of chronic adult periodontitis in AIDS patients.     

牙龈卟啉单胞菌感染宿主细胞后对其凋亡调控研究进展

牙龈卟啉单胞菌（Porphyromonas gingivalis）是证据充分的重要的牙周致病菌之一,参与牙周炎的发生、发展及牙周组织的破坏过程。以往研究表明,牙周感染与全身健康及系统疾病均有着密切关系。P. gingivalis感染多种宿主细胞后,通过干扰相关信号传导及蛋白表达,对宿主细胞凋亡进行调控。现就P. gingivalis感染宿主细胞后对其凋亡调控研究进展做一综述。     

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??Porphyromonas gingivalis?? a gram-negative obligate anaerobic bacterium??has been proved to be one of the main periodontal pathogens?? participating in development of periodontitis and process of periodontal tissue destruction. Previous studies have demonstrated the close relationship between periodontal infection and systemic health and diseases. P. gingivalis infection on host cells mediated cell apoptosis by interfering with cell signal transduction and expression of related proteins. This article reviews research progress of P. gingivalis infection on mediation of apoptosis of host cells.     

Apoptosis in chronic adult periodontitis analyzed by in situ DNA breaks, electron microscopy, and immunohistochemistry

BACKGROUND: Apoptosis is an evolutionary form of physiological cell death. Previous studies suggest that apoptosis is involved in the pathogenesis of periodontal diseases. Therefore, we studied the apoptotic events in the gingival tissue of chronic adult periodontitis patients. METHODS: Gingival tissue biopsies from 22 patients with chronic adult periodontitis and from 11 healthy controls were obtained. Criteria for patient inclusion in the periodontitis group were a minimum of 14 natural teeth, excluding third molars, with at least 10 posterior teeth; 5 to 6 sites with probing depth > or = 5 mm; attachment loss > or = 3 mm; and extensive radiographic bone loss. The control group included healthy subjects with no prior history of periodontal disease. Apoptosis was determined using the terminal TdT-mediated dUTP-biotin nick end labeling (TUNEL) technique; electron microscopic analysis; and expression of Caspase-3, Fas, FasL, Bcl-2, and p53 by immunohistochemistry. RESULTS: TUNEL-positive cells and cells exhibiting chromatin condensation by electron microscopy were observed in the inflammatory infiltrate of biopsies obtained from periodontitis patients. Most of the TUNEL-positive cells belonged to neutrophil cell populations as they were stained with anti-myeloperoxidase. Positive staining for active-caspase 3, Fas, FasL, and p53 was only observed in the inflammatory infiltrate from periodontitis biopsies, whereas Bcl-2 cells were present in both periodontitis patients and healthy controls. CONCLUSIONS: Our findings establish that apoptosis is induced in the periodontal tissue by host and microbial factors and support the hypothesis that apoptotic mechanisms could be implicated in the inflammatory process associated with gingival tissue destruction observed in adult periodontitis patients.     

2007年辽宁省2型糖尿病患者牙周炎患病情况调查

目的 调查2007年辽宁省2型糖尿病患者牙周炎患病率,分析此人群牙周炎发病的影响因素,以期为牙周病的预防和研究提供依据.方法 选取辽宁省2007年糖尿病流行病学研究普查出的2型糖尿病患者182例,进行问卷调查,并检查6颗指数牙的牙周探诊深度(probing depth,PD)、附着丧失(attachment loss,AL)、龈沟出血指数(sulcus bleeding index,SBI)、简化口腔卫生指数(simplified oral hygiene index,OHI-S)的情况.体格和生化指标检查:测量身高、体质量、血压、心电图等.患者检查前24 h内禁食油腻食物,取上午8:00到10:00的空腹静脉血及口服75 g葡萄糖后2 h的静脉血,全自动血生化仪分析血清生化指标.结果 182例2型糖尿病患者牙周炎患病率为96.7%(176/182),其中轻度牙周炎20例,占11.0%,中、重度牙周炎156例,占85.7%.182例患者共检测6552个位点,平均PD为(2.92±0.67)mm,平均AL为(2.87±1.31)mm.男性患者的平均AL、SBI明显大于女性,口腔卫生状况较女性差;城市患者的SBI重于农村患者,农村患者的口腔卫生状况较城市患者差.57.1%(104/182)的患者至少有1颗牙脱落(不包括第三磨牙).年龄、性别、吸烟、城乡差异、糖耐量实验2 h血糖与牙周指标具有相关性(P＜0.05或P＜0.01).结论 2007年辽宁省2型糖尿病患者牙周炎患病率高且牙周破坏严重,口腔卫生教育及防病、治病意识亟待加强.