M1型、M2型巨噬细胞极化相关因子在重度慢性牙周炎患者牙龈组织中的表达及其临床意义

Expression and clinical significance of factors related to M1 and M2 polarization of macrophages in gingival tissues of patients with severe chronic periodontitis

摘要:

目的 探讨M1型、M2型巨噬细胞极化相关因子在重度慢性牙周炎（CP）患者牙龈组织中的表达及其临床意义。方法 选取2018年10月—2021年6月柳州市人民医院收治的168例CP患者。根据病情严重程度分为轻度组、中度组和重度组，分别有27例、72例和69例。另选取同期该院53例牙龈组织健康者作为对照组。通过实时荧光定量聚合酶链反应检测研究对象牙龈组织中M1型巨噬细胞极化相关因子诱导型一氧化氮合酶（iNOS）、信号传导及转录激活因子1（STAT1）及M2型巨噬细胞极化相关因子精氨酸酶1（Arg1）、STAT6的表达。分析不同人群牙龈组织中M1型、M2型巨噬细胞极化相关因子mRNA相对表达量，分析CP患者病情严重程度的影响因素，分析M1型、M2型巨噬细胞极化相关因子对CP患者病情严重程度的诊断效能。结果 轻度组、中度组和重度组牙龈组织中iNOS、STATl、Arg1、STAT6 mRNA相对表达量较对照组高，中度组和重度组较轻度组高，重度组较中度组高。轻中度组与重度组性别、年龄、体质量指数、高血压、糖尿病、冠心病、高脂血症、吸烟史、饮酒史、偏侧咀嚼、巴氏刷牙、刷牙≥ 2次/d、刷牙≥ 3 min/次、夜磨牙症、血清钙、SBI、PLI比较，差异均无统计学意义（P >0.05）。重度组TNF-β、IL-10、IL-17及iNOS、STATl、Arg1、STAT6 mRNA相对表达量高于轻中度组。多因素Logistic逐步回归分析结果显示：IL-10 ［O^R=3.873（95% CI：1.594，9.412）］、IL-17 ［O^R=4.267（95% CI：1.756，10.371）］及iNOS ［O^R=4.092（95% CI：1.684，9.944）］、Arg1［O^R=3.651（95% CI：1.502，8.873）］、STAT6 ［O^R=3.644（95% CI：1.499，8.856）］mRNA相对表达量是影响CP患者病情严重程度的危险因素（P <0.05）。受试者工作特征曲线结果显示，iNOS、Arg1、STAT6 mRNA联合预测CP患者病情严重程度的敏感性、特异性和曲线下面积最高，分别为81.16%（95% CI：0.696，0.892）、90.91（95% CI：0.830，0.955）、0.908（95% CI：0.854，0.963）。结论 M1型、M2型巨噬细胞极化相关因子iNOS、Arg1、STAT6与CP患者病情严重程度有关，3者联合评估CP患者病情严重程度效能良好。

Abstract:

Objective To explore the expression and clinical significance of factors related to M1 and M2 polarization of macrophages in gingival tissues of patients with severe chronic periodontitis (CP).Methods A total of 168 CP patients admitted to the hospital from October 2018 to June 2021 were included, and 27, 72 and 69 patients were subdivided into mild CP group, moderate CP group, and severe CP group, respectively according to the disease severity. Another 53 individuals with healthy gingival tissues in the hospital during the same period were selected as the control group. Real-time fluorescent quantitative PCR was used to detect the expressions of factors related to M1 polarization of macrophages, including inducible nitric oxide synthase (iNOS) and signal transducer and activator of transcription 1 (STAT1), and those related to M2 polarization of macrophages, including arginase I (Arg1) and STAT6. The relative mRNA expressions of these factors in gingival tissues were determined among different populations, and factors affecting the severity of CP were analyzed. The diagnostic efficacy of factors related to M1 and M2 polarization of macrophages in the severity of CP was also analyzed.Results The relative mRNA expressions of iNOS, STAT1, Arg1 and STAT6 in the gingival tissues of the mild CP group, moderate CP group, and severe CP group were higher than those of the control group (P < 0.05). Besides, the relative mRNA expressions of iNOS, STAT1, Arg1 and STAT6 in the gingival tissues of the moderate and severe CP groups were higher than those of the mild CP group, while they were even higher in the gingival tissues of the severe CP group than those of the moderate CP group (P < 0.05). There was no difference in the gender, age, BMI, the presence of hypertension, diabetes mellitus, coronary heart disease, hyperlipidemia and bruxism, the history of smoking and alcohol, unilateral mastication, use of Bass technique of toothbrushing, the frequency of toothbrushing no less than twice a day, the duration of toothbrushing no less than 3 minutes per time, the serum level of calcium, SBI and PLI between the mild to moderate CP group and the severe group (P > 0.05). The levels of TNF-β, IL-10 and IL-17 as well as the relative mRNA expressions of iNOS, STAT1, Arg1 and STAT6 in the severe CP group were higher than those in the mild to moderate CP group (P < 0.05). Multivariable Logistic regression analysis showed that levels of IL-10 [O^R = 3.873, (95% CI: 1.594, 9.412) ] and IL-17 [O^R = 4.267, (95% CI: 1.756, 10.371) ], and the relative mRNA expressions of iNOS [O^R = 4.092, (95% CI: 1.684, 9.944) ], Arg1 [O^R = 3.651, (95% CI: 1.502, 8.873) ] and STAT6 [O^R = 3.644, (95% CI: 1.499, 8.856) ] were factors affecting the severity of CP patients (P < 0.05). The results of ROC curve analysis revealed that the combination of relative mRNA expressions of iNOS, Arg1 and STAT6 achieved the highest sensitivity, specificity and the AUC for predicting the severity of CP patients, being 81.16% (95% CI: 69.58%, 89.21%), 90.91% (95% CI: 83.01%, 95.50%), and 0.908 (95% CI: 0.854, 0.963), respectively.Conclusions Factors related to M1 and M2 polarization of macrophages including iNOS, Arg1 and STAT6 are associated with the severity of CP patients, and the combination of the three factors is effective for evaluating the severity of CP patients.