高压氧对糖尿病型牙周炎牙龈细胞凋亡及凋亡因子表达的调控作用研究

Regulatory effects of hyperbaric oxygen on cell apoptosis and apoptosis factors expression in gingival cells of diabetes-associated periodontitis

目的探讨高压氧(hyperbaric oxygen,HBO)对糖尿病型牙周炎(diabetes-associated periodontitis,DAP)牙龈组织细胞凋亡及凋亡因子表达的调控作用及机制。方法选择DAP患者60例,其中20例予以0.25 MPa HBO治疗2周,20例予以牙周洁治和根面平整(scaling and root planning,SRP)治疗,20例予以HBO+SRP治疗,在治疗前和治疗后1个月测定PLI、SBI、GI、PD、AL,用HE染色、TUNNEL染色和透射电镜观察牙龈细胞凋亡,免疫组化(IHC)检测牙龈组织IL-1β、TNF-α、PGE2、Caspase3表达。结果治疗后1个月时HBO组、SRP组、HBO+SRP组SBI、GI、PD比治疗前明显降低;牙龈上皮层和固有层细胞凋亡数量和凋亡细胞阳性表达指数明显降低,牙龈组织IL-1β、TNF-α、PGE2及Caspase3表达明显降低。治疗后1个月时HBO+SRP组SBI、GI、PD、AL及牙龈细胞凋亡指数与HBO组和SRP组相比均有显著性差异;牙龈组织IL-1β、TNF-α、PGE2、Caspase3表达比HBO组和SRP组降低。IL-1β、TNF-α、PGE2主要阳性细胞为巨噬细胞和淋巴细胞,Caspase3阳性细胞为上皮基底细胞、棘细胞。结论HBO可显著降低DAP患者牙龈组织中IL-1β、TNF-α、PGE2含量,减少牙龈组织中Caspase3表达,抑制细胞凋亡,HBO联合SRP疗效更好,可维持1个月以上。

Objective To investigate the regulatory effects of hyperbaric oxygen(HBO)on cell apoptosis and apoptosis factors expression of diabetes-associated periodontitis(DAP).Methods Sixty cases of DAP were divided into 3 groups,20 cases in each group.The first group was treated with 0.25 MPa hyperbaric oxygen(HBO)for 2 weeks;the second group was treated with periodontal scaling and root planning(SRP);the third group was treated with HBO combining SRP.Periodontal clinical indicators(PLI,SBI,GI,PD,AL)in each group before and one month after treatment were measured.The apoptosis of gingival cells was observed by HE staining,TUNNEL staining and transmission electron microscopy.The IL-1β,TNF-α,PGE2 and Caspase3 expression in gingival tissues were detected by immunohistochemistry(IHC).Results The periodontal indices of SBI,GI,PD significantly decreased in HBO group,SRP group and HBO+SRP group at one month after treatment than pre-treatment.Cell apoptosis and percentage of cell apoptosis in gingival epithelium and lamina propria decreased significantly.The expressions of IL-1β,TNF-α,PGE2 and Caspase3 in gingival at one month after treatment significantly decreased than pre-treatment.There were significantly differences between the clinical indices(SBI,GI,PD,AL)and the percentage of apoptotic cells.The expressions of IL-1β,TNF-α,PGE2 and Caspase3 in gingival of HBO+SRP group were significantly lower than those of HBO group and SRP group.The positive expressions of IL-1β,TNF-α,PGE2 were macrophages and lymphocytes,and the positive expressions of Caspase3 were epithelial basal cells and spinous cells.Conclusion HBO could reduce the expression of IL-1β,TNF-α,PGE2 and Caspase3 in gingival to reduce cell apoptosis.HBO combined with SRP displays better efficacy which could maintain for more than one month.