Th1/Th2、M1-M2与2型糖尿病慢性牙周炎病变程度及预后的关系

Relationship of Th1/Th2 and M1-M2 with severity and prognosis of patients with type 2 diabetes mellitus complicated with chronic periodontitis

目的探讨2型糖尿病(T2DM)合并慢性牙周炎(CP)患者辅助性T细胞1型/辅助性T细胞2型(Th1/Th2)失衡及巨噬细胞M1-M2极化,以及其与牙周炎病变程度和治疗预后的关系。方法采用前瞻性研究方法,纳入2022年1月至2023年6月在宝鸡市人民医院口腔科治疗的80例T2DM合并CP患者作为研究对象,根据CP严重程度分为轻度组(57例)和中重度组(23例),治疗后根据预后分为预后良好组(33例)和预后差组(47例)。轻度组男28例、女29例,年龄(48.00±6.03)岁,T2DM病程(3.54±1.15)年,CP病程(12.54±4.65)个月;中重度组男11例、女12例,年龄(50.48±6.65)岁,T2DM病程(3.79±1.55)年,CP病程(12.71±4.76)个月。采用流式细胞术评估Th1/Th2细胞比例,通过实时定量PCR分析牙周组织中M1型与M2型巨噬细胞标志物的表达。采用Pearson相关性分析Th1/Th2细胞比例、M1/M2极化状态与牙周炎临床参数及预后之间的相关性。统计学方法采用t检验、χ^(2)检验。结果中重度组Th1、Th1/Th2细胞比例及肿瘤坏死因子(TNF)-α、白细胞介素(IL)-2、IL-4、IL-6、IL-10、IL-1β、IL-12、IL-17、TNF-β、信号转导和转录激活因子1(STAT1)、诱导型一氧化氮合酶(iNOS)、信号转导和转录激活因子6(STAT6)mRNA、精氨酸酶1(Arg1)mRNA指标均高于轻度组[(2.62±0.53)%比(1.62±0.41)%、(2.47±0.53)比(1.21±0.27)、(51.83±6.25)ng/L比(36.74±5.47)ng/L、(27.54±50.00)μg/L比(14.58±4.65)μg/L、(72.63±8.95)ng/L比(46.68±6.57)ng/L、(41.85±2.61)ng/L比(37.08±3.54)ng/L、(4.76±1.13)ng/L比(3.87±1.16)ng/L、(23.10±5.86)μg/L比(16.78±2.54)μg/L、(11.76±5.37)ng/L比(9.16±2.16)ng/L、(12.78±2.14)μg/L比(10.54±1.63)μg/L、(1370.0±160.0)μg/L比(910.0±140.0)μg/L、(800.0±180.0)μg/L比(260.0±110.0)μg/L、(930.0±190.0)μg/L比(430.0±110.0)μg/L、(760.0±130.0)μg/L比(560.0±90.0)μg/L、(710.0±120.0)μg/L比(490.0±80.0)μg/L],Th2细胞比例及干扰素-γ(IFN-γ)、IL-23水平均低于轻度组[(1.06±0.37)%比(1.34±0.41)%、(4.21±0.65)ng/L比(4.85±0.67)ng/L、(6.84±0.83)ng/L比(14.65±1.81)ng/L],差异均有统计学意义(t=9.05、14.06、10.71、10.69、14.35、5.84、3.12、6.76、3.10、5.07、12.76、16.37、14.73、7.87、9.57、2.84、3.95、19.81,均P<0.05)。患者牙周炎病变程度与Th1、Th1/Th2、TNF-α、IL-2、IL-4、IL-6、IL-10、IL-1β、IL-12、IL-17、TNF-β、STAT1、iNOS、STAT6 mRNA、Arg1 mRNA均呈正相关(均P<0.01);与Th2、INF-γ、IL-23均呈负相关(均P<0.01)。预后良好组Th1、Th1/Th2细胞比例及TNF-α、IL-1β水平均低于预后差组[(1.31±0.21)%比(2.62±0.75)%、(0.94±0.21)比(2.70±0.48)、(34.81±4.51)ng/L比(55.23±7.31)ng/L、(15.84±2.89)μg/L比(24.56±4.74)μg/L],Th2细胞比例高于预后差组[(1.39±0.24)%比(0.97±0.37)%],差异均有统计学意义(t=9.75、19.74、14.24、9.40、5.72,均P<0.05)。结论T2DM合并CP患者Th1/Th2失衡及M1-M2极化状态变化与牙周病变程度和治疗预后密切相关,调节免疫反应对改善T2DM患者牙周病变和预后具有潜在的临床意义。

Objective To explore the imbalance of Th1/Th2 cell-mediated immunity and the changes in macrophage M1-M2 polarization in patients with type 2 diabetes mellitus(T2DM)complicated with chronic periodontitis(CP)and their relationship with the degree of periodontal lesions and treatment prognosis.Methods Eighty patients with T2DM and CP who were treated at Department of Stomatology,Baoji People's Hospital from January 2022 to June 2023 were enrolled in this prospective study.Based on the severity of CP,the patients were divided into a mild group(57 cases)and a moderate-severe group(23 cases).According to the prognosis after the treatment,they were divided into a good prognosis group(33 cases)and a poor prognosis group(47 cases).There were 28 males and 29 females in the mild group;they were(48.00±6.03)years old;their T2DM duration was(3.54±1.15)years;their CP duration was(12.54±4.65)months.There were 11 males and 12 females in the moderate-severe group;they were(50.48±6.65)years old;their T2DM duration was(3.79±1.55)years;their CP duration was(12.71±4.76)months.The flow cytometry was used to assess the ratio of Th1/Th2 cells.The real-time quantitative PCR was employed to analyze the expression of M1 and M2 macrophage markers in periodontal tissue.The Pearson correlation analysis was performed to evaluate the correlations of Th1/Th2 ratio and M1/M2 polarization with the clinical parameters of periodontitis and prognosis.t andχ^(2) tests were used.Results The proportion of Th1 cells,Th1/Th2,and levels of tumor necrosis factor(TNF)-α,interleukin(IL)-2,IL-4,IL-6,IL-10,IL-1β,IL-12,IL-17,TNF-β,inducible nitric oxide synthase(iNOS),signal transducer and activator of transcription 6(STAT6)mRNA,and arginase 1(Arg1)mRNA in the moderate-severe group were higher than those in the mild group[(2.62±0.53)%vs.(1.62±0.41)%,(2.47±0.53)vs.(1.21±0.27),(51.83±6.25)ng/L vs.(36.74±5.47)ng/L,(27.54±50.00)μg/L vs.(14.58±4.65)μg/L,(72.63±8.95)ng/L vs.(46.68±6.57)ng/L,(41.85±2.61)ng/L vs.(37.08±3.54)ng/L,(4.76±1.13)ng/L vs.(3.87±1.16)ng/L,(23.10±5.86)μg/L vs.(16.78±2.54)μg/L,(11.76±5.37)ng/L vs.(9.16±2.16)ng/L,(12.78±2.14)μg/L vs.(10.54±1.63)μg/L,(1370.0±160.0)μg/L vs.(910.0±140.0)μg/L,(800.0±180.0)μg/L vs.(260.0±110.0)μg/L,(930.0±190.0)μg/L vs.(430.0±110.0)μg/L,(760.0±130.0)μg/L vs.(560.0±90.0)μg/L,and(710.0±120.0)μg/L vs.(490.0±80.0)μg/L];the proportion of Th2 cells and the levels of interferonγ(IFN-γ)and IL-23 in the in the moderate-severe group were lower than those in the mild group[(1.06±0.37)%vs.(1.34±0.41)%,(4.21±0.65)ng/L vs.(4.85±0.67)ng/L,and(6.84±0.83)ng/L vs.(14.65±1.81)ng/L];there were statistical differences(t=9.05,14.06,10.71,10.69,14.35,5.84,3.12,6.76,3.10,5.07,12.76,16.37,14.73,7.87,9.57,2.84,3.95,and 19.81;all P<0.05).The severity of CP positively correlated with Th1,Th1/Th2,TNF-α,IL-2,IL-4,IL-6,IL-10,IL-1β,IL-12,IL-17,IL-23,TNF-β,STAT1,iNOS,STAT6 mRNA,and Arg1 mRNA(all P<0.01),and negatively with Th2,INF-γ,and IL-23(all P<0.01).The proportion of Th1 cells,Th1/Th2,and levels of TNF-αand IL-1βin the good prognosis group were lower than those in the poor prognosis group[(1.31±0.21)%vs.(2.62±0.75)%,(0.94±0.21)vs.(2.70±0.48),(34.81±4.51)ng/L vs.(55.23±7.31)ng/L,(15.84±2.89)μg/L vs.(24.56±4.74)μg/L];the proportion of Th2 cells in the good prognosis group was higher than that in the poor prognosis group[(1.39±0.24)%vs.(0.97±0.37)%];there were statistical differences(t=9.75,19.74,14.24,9.40,and 5.72;all P<0.05).Conclusion The imbalance of Th1/Th2 and changes in M1-M2 polarization status in patients with T2DM and CP are closely related to the degree of periodontal lesions and treatment prognosis,indicating that modulating immune responses may have potential clinical significance in improving periodontal lesions and prognosis of patients with T2DM.