吸烟者和正常牙龈上皮细胞凋亡的对比研究(1)
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【摘要】 目的 对比吸烟者和未吸烟者的牙龈上皮细胞凋亡的情况不同,评价吸烟对牙龈上皮细胞凋亡的影响并探讨其可能机制。方法 选择21例行冠延长术的男性患者,年龄为30~45岁,无相关系统疾病和明显的牙龈疾病。无吸烟史10人作对照,切取的牙龈上皮经新鲜配置4%多聚甲醛固定、 脱水、石蜡包埋,TUNEL(TdT- mediated - dUTP nick end labeling, TUNEL) 法原位细胞凋亡检测牙龈上皮的细胞凋亡情况。Newman-Keuls法计算各吸烟组与对照组及各吸烟组之间的差异,取α=0.05水准。SP免疫组化法对凋亡抑制基因Bcl-2的表达进行组织定位。结果 各吸烟组与对照组都存在显著差异(P<0.01);轻度吸烟组和重度吸烟组间也存在显著性差异(P<0.01)。Bcl-2在吸烟者的牙龈上皮的表达较对照组减弱。结论 吸烟促进牙龈上皮细胞凋亡的发生,干扰牙龈上皮的正常代谢。【关键词】 吸烟;细胞凋亡;原位细胞凋亡检测;牙龈上皮
Comparison between smokers and non-smokers on apoptosis in gingival epithelium
XUE Lan-de,XU Dong-mei,LU ji-kun,et al.
Department of Periodontology,School of Stomatology, Shandong University. Jinan 250012,China
【Abstract】 Objective To compare the different apoptosis in gingival epithelium between smokers and non-smokers. Methods 21 healthy male patients of crown lengthening surgery without systemic or distinct gingival diseases, the age ranged from 30 to 45years old. Ten normal gingival epithelium were chosen as control. Apoptosis was identified by the terminal deoxy-transferase (TdT)-mediated dUTP-biotin nick end labeling (TUNEL) method. Differences among the three groups were analyzed using Student-Newman-Keuls (SNK).The criteria for statistical significance were accepted at the probability level P<0.05. SP immunohistochemical method was used to detect the expression of Bcl-2 protein. Results Apoptosis in human gingival epithelium were significant difference between smokers and non-smokers (P<0. 01). The expression of Bcl-2 was weaker in gingival epithelium of smokers. Conclusion We conclude that exposure to cigarette smoke can increase apoptosis in gingival epithelium and interfere with normal metabolism.
【Key words】 Smoking; Apoptosis; TUNEL; Gingiva epithelium
DOI:10.3760/cma.j.issn 1673-8799.2010.11.18
作者单位:250001济南市口腔医院(薛兰德 徐冬梅 路吉坤 于西佼);山东大学口腔医学院(李纾)
大量研究证实,吸烟状况与牙周和口腔黏膜疾病的发生存在明显的相关性[1],吸烟状况可作为牙周和口腔黏膜疾病发生危险度的一个关键指标[2,3]。但对于吸烟和牙龈上皮细胞凋亡的关系少有报道,本研究采用TUNEL法检测不同吸烟情况者牙龈上皮细胞凋亡的情况,同时观察凋亡抑制基因Bcl-2的表达改变,探讨吸烟影响牙龈上皮细胞凋亡的可能机制。
1 材料和方法
1.1 病例选择和标本制备
选取21例具有不同吸烟史的无系统疾病的牙冠延长术男性患者,对患者的年龄30～45、吸烟史、日平均吸烟支数、口腔健康状况及牙位等详细记录,根据吸烟史和日平均吸烟支数将所有患者分为Ⅰ轻度(吸烟史<5年,日均支数<10)9例、Ⅱ重度(5年≤吸烟史,10 ≤日均支数)12例,选取正常的牙龈上皮组织10例作为对照组。所有组织标本均经新鲜配置4%多聚甲醛固定4℃2 h,梯度乙醇脱水,石蜡包埋,制备5μm切片 ......
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