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毛细支气管炎是婴幼儿时期常见的下呼吸道感染性疾病,可由多种病毒感染引起,如呼吸道合胞病毒(RSV)、鼻病毒、腺病毒等[1],未来可能发展为支气管哮喘。其中仅最常见的RSV感染就可造成每年3 310万次急性发病、320万次住院治疗和11.82万例患儿死亡[2],且缺乏有效的特异性治疗措施,危害极大。毛细支气管炎发病机制至今未明[3],目前研究最多的是免疫学发病机制,其中Th17/Treg失衡参与哮喘的发病,但是否参与关于毛细支气管炎的研究相对较少。调节性T细胞(Treg)是免疫抑制T细胞,可分泌白细胞介素(IL)-10和IL-35等免疫抑制因子,对多种免疫细胞发挥抑制作用。辅助T淋巴细胞(Th17)是T淋巴细胞亚群的一种,属于促炎细胞,能够分泌IL-17等发挥作用,现有研究发现Th17可能参与毛细支气管炎的发病机制和免疫过程[4]。本研究通过检测毛细支气管炎患儿体内Th17、Treg以及相关细胞因子的水平,探讨Th17/Treg及细胞因子在疾病发生发展中的作用,为临床病情预判和治疗寻找靶点提供依据。
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病例组各组Th17、Th17/Treg、IL-6、IL-17较对照组升高,而Treg、IL-6、IL-10较对照组降低,差异均有统计学意义(P<0.05);性别、年龄、白细胞计数、中性粒胞计数、淋巴细胞计数、嗜酸细胞计数、CRP、PCT、HBP各组差异均无统计学意义(P>0.05)(见表 1~2)。
分组 n 年龄(x±s)/月 男 女 白细胞计数(x±s)/(×109/L) 中性粒细胞计数/ (×109/L) 淋巴细胞计数(x±s)/(×109/L) 嗜酸细胞计数/ (×109/L) 对照组 30 4.38±2.84 22 8 8.86±2.49 3.18(1.98, 4.53) 4.16±1.82 0.04(0, 0.14) 轻症组 42 4.89±3.06 30 12 9.49±3.43 2.72(1.60, 4.22) 4.65±1.31 0.01(0, 0.07) 中症组 21 4.29±3.04 13 8 9.47±4.07 2.86(1.60, 6.08) 4.38±1.97 0.01(0, 0.03) 重症组 20 3.34±2.65 15 5 7.75±2.90 2.14(0.81, 3.32) 4.17±1.66 0.04(0, 0.07) F — 1.87 1.08* 1.46 5.48△ 0.65 5.79△ P — >0.05 >0.05 >0.05 >0.05 >0.05 >0.05 *示χ2值;△示uc值 表 1 对照组和病例组之间一般资料比较[M(P25, P75)]
分组 n Th17/% Treg/% Th17/Treg IL-17/(pg/mL) IL-6/(pg/mL) IL-10/(pg/mL) IL-35(x±s)/ (pg/mL) 对照组 30 1.00(0.72, 1.35) 6.33(5.21, 7.34) 0.17(0.13, 0.21) 17.41(14.22, 22.55) 2.12(1.51, 2.66) 4.96(4.19, 5.76) 79.38±12.58 轻症组 42 1.27(1.04, 1.44)* 5.27(4.87, 5.73)* 0.24(0.19, 0.27)* 20.60(19.23, 22.28)* 2.52(2.15, 2.97)* 4.52(3.68, 5.18)* 66.75±16.61* 中症组 21 1.23(0.83, 1.85)* 5.02(3.82, 5.86)* 0.27(0.17, 0.40)* 22.35(20.30, 27.35)* 2.36(1.86, 3.69)* 3.98(3.11, 6.18) 63.33±10.14* 重症组 20 1.92(1.58, 2.24)*△# 4.45(3.32, 5.23)*△ 0.40(0.33, 0.68)*△# 31.25(28.35, 35.41)*△# 3.98(3.23, 5.19)*△# 3.47(2.69, 4.36)*△ 57.08±15.92*△ uc — 28.43 22.04 42.29 47.95 29.89 14.76 10.83◇ P — < 0.01 < 0.01 < 0.01 < 0.01 <0.01 <0.01 <0.01 注:与对照组比较*P<0.05;与轻症组比较△P<0.05;与中症组比较#P<0.05。◇示F值 表 2 对照组和病例组之间各项实验指标比较[M(P25, P75)]
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重症组Th17、Th17/Treg、IL-6、IL-17较中症组和轻症组均增高,Treg、IL-10、IL-35较轻症组降低,差异均有统计学意义(P<0.05);中症组和轻症组间差异无统计学意义(P>0.05);CRP、PCT、HBP、住院时间、早产率、剖宫产率、喂养方式各组差异均无统计学意义(P>0.05)(见表 2~3)。
分组 n CRP/(mg/L) PCT/(ng/mL) HBP/(μg/L) 住院时间(x±s)/d 早产[n; 百分率(%)] 剖宫产[n; 百分率(%)] 喂养方式[n; 百分率(%)] 人工喂养 混合喂养 母乳喂养 轻症组 42 1(0.5, 3.63) 0.33(0.15, 0.73) 8.65(6.51, 14.05) 8.55±3.03 5(11.90) 22(52.38) 18(42.86) 8(19.05) 16(38.10) 中症组 21 1.5(0.61, 7.11) 0.24(0.14, 0.80) 8.71(6.05, 13.89) 8.10±2.47 0(0.00) 9(42.86) 7(33.33) 1(0.05) 13(61.90) 重症组 20 1.25(0.48, 5.00) 0.19(0.12, 0.70) 9.04(6.39, 17.35) 10.05±2.42 2(10.00) 7(35.00) 6(30.00) 5(25.00) 9(45.00) uc — 1.18 2.14 0.49 2.91* 2.65△ 1.67△ 1.15△ 3.28△ 3.20△ P — >0.05 >0.05 >0.05 >0.05 >0.05 >0.05 >0.05 >0.05 >0.05 *示F值;△示χ2值 表 3 病例组间临床资料和实验指标比较[M(P25, P75)]
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病例组Th17与Th17/Treg、IL-17、IL-6呈正相关,与IL-35呈负相关;Treg与IL-10、IL-35呈正相关,与Th17/Treg负相关; Th17/Treg与IL-17、IL-6之间呈正相关,与IL-10、IL-35负相关;IL-17与IL-6呈正相关,与IL-35呈负相关(P<0.05~P<0.01)(见表 4)。
变量 Th17 Treg Th17/Treg IL-17 IL-6 IL-10 Treg r -0.20 P >0.05 Th17/Treg r 0.82 -0.62 P < 0.01 < 0.01 IL-17 r 0.41 -0.13 0.35 P < 0.01 >0.05 <0.01 IL-6 r 0.30 -0.16 0.27 0.33 P < 0.01 >0.05 <0.05 <0.01 IL-10 r -0.14 0.23 -0.21 0.09 -0.09 P >0.05 <0.05 < 0.05 >0.05 >0.05 IL-35 r -0.33 0.36 -0.36 -0.22 -0.07 0.03 P <0.01 <0.01 <0.01 <0.05 >0.05 >0.05 表 4 Th17、Treg及相关细胞因子之间相关性分析
变量 B SE Waldsχ2 P OR(95%CI) Th17/Treg 3.511 1.640 4.59 <0.05 33.476(1.346~832.583) IL-17 6.264 2.267 7.64 <0.01 525.085(6.176~44 640.125) IL-6 3.163 1.558 4.12 <0.05 23.641(1.117~500.598) IL-10 -3.405 1.982 2.95 >0.05 0.033(0.001~1.616) 常量 -16.347 5.995 7.43 <0.01 0.000(-) 表 5 重症毛细支气管炎危险因素logistic回归分析
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以重症、非重症为因变量,Th17/Treg、IL-17、IL-6、IL-10、IL-35为自变量,进行logistic回归分析, 结果显示Th17/Treg、IL-17和IL-6是毛细支气管炎的重症危险因素(P<0.05~P<0.01)(见表 5)。
Th17/Treg失衡与毛细支气管炎发病关系研究
Imbalance of Th17/Treg in the pathogenesis of bronchiolitis
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摘要:
目的探讨毛细支气管炎患儿外周血辅助T淋巴细胞17(Th17)/调节性T淋巴细胞(Treg)和相关细胞因子的水平与本病发展的关系。 方法选取毛细支气管炎患儿83例为病例组,分为轻症组(42例)、中症组(21例)和重症组(20例),同期健康体检的儿童30名为对照组。采用流式细胞学检测外周血辅助T细胞17(Th17)和调节性T细胞(Treg)占CD4+T细胞的比值,应用ELISA方法检测血清白细胞介素6(IL-6)、IL-10、IL-17和IL-35的浓度,比较各组间上述指标差异。 结果病例组Th17细胞比值及其分泌的促炎因子IL-17、IL-6较对照组明显增高(P<0.05),而Treg细胞比值及其分泌的抑炎因子IL-10、IL-35则明显降低,重症组愈加显著(P<0.05)。Pearson相关分析显示Th17与Th17/Treg、IL-17、IL-6呈正相关;Treg与IL-10、IL-35呈正相关,与Th17/Treg负相关;Th17/Treg、Th17和IL-17与IL-35负相关(P<0.05~P<0.01)。logistic回归分析显示Th17/Treg、IL-7和IL-6是重症毛细支气管炎的危险因素(P<0.05~P<0.01)。 结论Th17/Treg失衡在毛细支气管炎发病中起着重要的作用,Th17/Treg、IL-17和IL-6是重症毛细支气管炎的危险因素。 Abstract:ObjectiveTo investigatethe the levels of helper T lymphocyte 17(Th17)/regulatory T lymphocyte(Treg) and related cytokines in children with bronchiolitis, and its correlation with the progress of disease. MethodsEighty-three children with bronchiolitis were divided into the light disease group(42 cases), medium disease group(21 cases) and severe disease group(20 cases), and 30 healthy physical examination children at the same time were set as the control group.The ratio of Th17 and Treg to CD4+T cells were detected using flow cytometry.The serum concentrations of interleukin(IL)-6, IL-10, IL-17 and IL-35 were detected using ELISA, and the differences of above indicators were analyzed among four groups. ResultsThe ratio of Th17 cells and levels of pro-inflammatory factors of IL-17 and IL-6 in case group were significantly higher than those in control group(P<0.05), while the ratio of Treg cells and production of anti-inflammatory factors of IL-10 and IL-35 in case group were significantly lower than those in control group(P<0.05), especially in the severe group.The results of Pearson correlation analysis showed that the level of Th17 was positively correlated with levels of Th17/Treg, IL-17 and IL-6, and the level of Treg was positively correlated with levels of IL-10 and IL-35, and negatively correlated with Th17/Treg.The levels of Th17/Treg, Th17 and IL-17 were negatively correlated with IL-35(P<0.05 to P<0.01).The results of logistic regression analysis showed that the odds ratio of severe bronchiolitis to Th17/Treg, IL-17 and IL-6 were statistically significant(P<0.05 to P<0.01). ConclusionsThe imbalance of Th17/Treg plays an important role in the pathogenesis of bronchiolitis.Th17/Treg, IL-7 and IL-6 are the risk factors of severe bronchiolitis. -
表 1 对照组和病例组之间一般资料比较[M(P25, P75)]
分组 n 年龄(x±s)/月 男 女 白细胞计数(x±s)/(×109/L) 中性粒细胞计数/ (×109/L) 淋巴细胞计数(x±s)/(×109/L) 嗜酸细胞计数/ (×109/L) 对照组 30 4.38±2.84 22 8 8.86±2.49 3.18(1.98, 4.53) 4.16±1.82 0.04(0, 0.14) 轻症组 42 4.89±3.06 30 12 9.49±3.43 2.72(1.60, 4.22) 4.65±1.31 0.01(0, 0.07) 中症组 21 4.29±3.04 13 8 9.47±4.07 2.86(1.60, 6.08) 4.38±1.97 0.01(0, 0.03) 重症组 20 3.34±2.65 15 5 7.75±2.90 2.14(0.81, 3.32) 4.17±1.66 0.04(0, 0.07) F — 1.87 1.08* 1.46 5.48△ 0.65 5.79△ P — >0.05 >0.05 >0.05 >0.05 >0.05 >0.05 *示χ2值;△示uc值 表 2 对照组和病例组之间各项实验指标比较[M(P25, P75)]
分组 n Th17/% Treg/% Th17/Treg IL-17/(pg/mL) IL-6/(pg/mL) IL-10/(pg/mL) IL-35(x±s)/ (pg/mL) 对照组 30 1.00(0.72, 1.35) 6.33(5.21, 7.34) 0.17(0.13, 0.21) 17.41(14.22, 22.55) 2.12(1.51, 2.66) 4.96(4.19, 5.76) 79.38±12.58 轻症组 42 1.27(1.04, 1.44)* 5.27(4.87, 5.73)* 0.24(0.19, 0.27)* 20.60(19.23, 22.28)* 2.52(2.15, 2.97)* 4.52(3.68, 5.18)* 66.75±16.61* 中症组 21 1.23(0.83, 1.85)* 5.02(3.82, 5.86)* 0.27(0.17, 0.40)* 22.35(20.30, 27.35)* 2.36(1.86, 3.69)* 3.98(3.11, 6.18) 63.33±10.14* 重症组 20 1.92(1.58, 2.24)*△# 4.45(3.32, 5.23)*△ 0.40(0.33, 0.68)*△# 31.25(28.35, 35.41)*△# 3.98(3.23, 5.19)*△# 3.47(2.69, 4.36)*△ 57.08±15.92*△ uc — 28.43 22.04 42.29 47.95 29.89 14.76 10.83◇ P — < 0.01 < 0.01 < 0.01 < 0.01 <0.01 <0.01 <0.01 注:与对照组比较*P<0.05;与轻症组比较△P<0.05;与中症组比较#P<0.05。◇示F值 表 3 病例组间临床资料和实验指标比较[M(P25, P75)]
分组 n CRP/(mg/L) PCT/(ng/mL) HBP/(μg/L) 住院时间(x±s)/d 早产[n; 百分率(%)] 剖宫产[n; 百分率(%)] 喂养方式[n; 百分率(%)] 人工喂养 混合喂养 母乳喂养 轻症组 42 1(0.5, 3.63) 0.33(0.15, 0.73) 8.65(6.51, 14.05) 8.55±3.03 5(11.90) 22(52.38) 18(42.86) 8(19.05) 16(38.10) 中症组 21 1.5(0.61, 7.11) 0.24(0.14, 0.80) 8.71(6.05, 13.89) 8.10±2.47 0(0.00) 9(42.86) 7(33.33) 1(0.05) 13(61.90) 重症组 20 1.25(0.48, 5.00) 0.19(0.12, 0.70) 9.04(6.39, 17.35) 10.05±2.42 2(10.00) 7(35.00) 6(30.00) 5(25.00) 9(45.00) uc — 1.18 2.14 0.49 2.91* 2.65△ 1.67△ 1.15△ 3.28△ 3.20△ P — >0.05 >0.05 >0.05 >0.05 >0.05 >0.05 >0.05 >0.05 >0.05 *示F值;△示χ2值 表 4 Th17、Treg及相关细胞因子之间相关性分析
变量 Th17 Treg Th17/Treg IL-17 IL-6 IL-10 Treg r -0.20 P >0.05 Th17/Treg r 0.82 -0.62 P < 0.01 < 0.01 IL-17 r 0.41 -0.13 0.35 P < 0.01 >0.05 <0.01 IL-6 r 0.30 -0.16 0.27 0.33 P < 0.01 >0.05 <0.05 <0.01 IL-10 r -0.14 0.23 -0.21 0.09 -0.09 P >0.05 <0.05 < 0.05 >0.05 >0.05 IL-35 r -0.33 0.36 -0.36 -0.22 -0.07 0.03 P <0.01 <0.01 <0.01 <0.05 >0.05 >0.05 表 5 重症毛细支气管炎危险因素logistic回归分析
变量 B SE Waldsχ2 P OR(95%CI) Th17/Treg 3.511 1.640 4.59 <0.05 33.476(1.346~832.583) IL-17 6.264 2.267 7.64 <0.01 525.085(6.176~44 640.125) IL-6 3.163 1.558 4.12 <0.05 23.641(1.117~500.598) IL-10 -3.405 1.982 2.95 >0.05 0.033(0.001~1.616) 常量 -16.347 5.995 7.43 <0.01 0.000(-) -
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