Toll受体及下游基因和T细胞亚群在新生儿感染性疾病中的变化及相关性
|
摘要
目的分析Toll受体及下游基因和T细胞亚群在新生儿感染性疾病中的变化及关系。方法选取2017年10月至2018年7月于该院诊治的108例新生儿感染患儿(感染组)和30例健康新生儿(健康对照组)作为研究对象,采用实时荧光定量PCR(qPCR)和流式细胞仪检测并比较入组时两组外周血单个核细胞Toll受体基因(TLR2、TLR9)及下游基因(MyD88、TRAF6)的mRNA及其编码蛋白表达情况,同时检测并比较两组外周血中T淋巴细胞亚群分布情况(CD3~+、CD4~+、CD8~+T淋巴细胞和CD4~+/CD8~+);分析患儿TLR2/TLR9mRNA和MyD88/TRAF6mRNA及编码蛋白的相关性,并比较以上mRNA和编码蛋白在新生儿不同感染中的差异。结果与健康对照组比较,感染组TLR2、TLR9、MyD88和TRAF6mRNA及其编码蛋白均显著升高;感染组CD3~+T淋巴细胞明显升高,而CD4~+和CD8~+T淋巴细胞及CD4~+/CD8~+比值则明显下降,以上差异均有统计学意义(P <0.05)。经Spearman分析得知,TLR2mRNA与TLR9mRNA、TLR2mRNA与TRAF6mRNA之间无明显相关性(r=0.430、0.418,P=0.214、0.229);TLR2mRNA与MyD88mRNA、TLR9mRNA与TRAF6mRNA均呈明显正相关(r=0.858、0.915,P=0.037、0.000)。不同类型感染性疾病患儿TLR2、TLR9、MyD88及TRAF6mRNA比较差异有统计学意义(P<0.05),且感染性肺炎和坏死性小肠结肠炎最高;不同病原菌感染性疾病患儿TLR2、TLR9、MyD88及TRAF6mRNA比较差异有统计学意义(P<0.05),且以细菌性感染最高。结论 TLR2和TLR9在新生儿感染中高表达,尤以细菌性感染、感染性肺炎和坏死性小肠结肠炎中最为明显,且与下游靶基因MyD88及TRAF6明显相关,有望成为辅助新生儿感染诊疗的有效指标,有一定推广价值。
Objective To explore the changes and relationship of Toll receptor,its downstream genes and T cell subsets in neonatal infectious diseases.Methods One hundred and eight cases of neonatal infection(infection group)and 30 healthy neonates in this hospital from October 2017 to July 2018 were selected as the research subjects.The expressions of Toll receptor gene(TLR2/TLR9),downstream gene(MyD88/TRAF6)mRNA and their coding proteins in peripheral blood mononuclear cells were detected by quantitative real-time polymerase chain reaction and flow cytometry and compared between the two groups.The distribution situation of T lymphocyte subsets(CD3~+,CD4~+,CD8~+ lymphocytes,CD4~+/CD8~+)in peripheral blood was also detected and compared between the two groups.The correlation among TLR2/TLR9 mRNA,MyD88/TRAF6 mRNA and coding protein was analyzed,and the differences of the above mRNA and coding protein were compared among different neonatal infections.Results Compared with the healthy control group,the levels of TLR2,TLR9,MyD88 and TRAF6 mRNA and their coding proteins in the infection group were significantly increased;CD3~+T lymphocytes were significantly increased in the infection group,while CD4~+ and CD8~+ Tlymphocytes and CD4~+/CD8~+ ratio were significantly decreased,the above differences were statistically significant(P<0.05).The Spearman analysis showed that there was no significant correlation between TLR2 mRNA with TLR9 mRNA,TLR2 mRNA and TRAF6 mRNA(r=0.430,0.418;P=0.214,0.229);TLR2 mRNA was significantly correlated with MyD88 mRNA,TLR9 mRNA and TRAF6 mRNA(r=0.858,0.915;P=0.037,0.000).TLR2,TLR9,MyD88 and TRAF6 mRNA had statistical difference among newborns with different types of infectious diseases(P<0.05),moreover which were the highest in infectious pneumonia and necrotizing enterocolitis.TLR2,TLR9,MyD88 and TRAF6 mRNA had statistical difference among newborns with different pathogenic bacterial infectious diseases(P<0.05),moreover which were the highest in bacterial infection.Conclusion TLR2 and TLR9 are highly expressed in neonatal infections,especially significant in bacterial infections,infectious pneumonia and necrotizing enterocolitis,moreover which are significantly correlated with its downstream target genes MyD88 and TRAF6,expected to be the effective indicators in assisting diagnosis and treatment of neonatal infections and have a certain promotion value.
Objective To explore the changes and relationship of Toll receptor,its downstream genes and T cell subsets in neonatal infectious diseases.Methods One hundred and eight cases of neonatal infection(infection group)and 30 healthy neonates in this hospital from October 2017 to July 2018 were selected as the research subjects.The expressions of Toll receptor gene(TLR2/TLR9),downstream gene(MyD88/TRAF6)mRNA and their coding proteins in peripheral blood mononuclear cells were detected by quantitative real-time polymerase chain reaction and flow cytometry and compared between the two groups.The distribution situation of T lymphocyte subsets(CD3~+,CD4~+,CD8~+ lymphocytes,CD4~+/CD8~+)in peripheral blood was also detected and compared between the two groups.The correlation among TLR2/TLR9 mRNA,MyD88/TRAF6 mRNA and coding protein was analyzed,and the differences of the above mRNA and coding protein were compared among different neonatal infections.Results Compared with the healthy control group,the levels of TLR2,TLR9,MyD88 and TRAF6 mRNA and their coding proteins in the infection group were significantly increased;CD3~+T lymphocytes were significantly increased in the infection group,while CD4~+ and CD8~+ Tlymphocytes and CD4~+/CD8~+ ratio were significantly decreased,the above differences were statistically significant(P<0.05).The Spearman analysis showed that there was no significant correlation between TLR2 mRNA with TLR9 mRNA,TLR2 mRNA and TRAF6 mRNA(r=0.430,0.418;P=0.214,0.229);TLR2 mRNA was significantly correlated with MyD88 mRNA,TLR9 mRNA and TRAF6 mRNA(r=0.858,0.915;P=0.037,0.000).TLR2,TLR9,MyD88 and TRAF6 mRNA had statistical difference among newborns with different types of infectious diseases(P<0.05),moreover which were the highest in infectious pneumonia and necrotizing enterocolitis.TLR2,TLR9,MyD88 and TRAF6 mRNA had statistical difference among newborns with different pathogenic bacterial infectious diseases(P<0.05),moreover which were the highest in bacterial infection.Conclusion TLR2 and TLR9 are highly expressed in neonatal infections,especially significant in bacterial infections,infectious pneumonia and necrotizing enterocolitis,moreover which are significantly correlated with its downstream target genes MyD88 and TRAF6,expected to be the effective indicators in assisting diagnosis and treatment of neonatal infections and have a certain promotion value.
引文
[1]宓秀菊,赵淑云,王敬彩,等.2014年度新生儿感染流行病学调查研究[J].中华医院感染学杂志,2015,25(17):4039-4040.
[2]KAN B,RAZZAGHIAN H R,LAVOIE P M.An immunological perspective on neonatal sepsis[J].Trends Mol Med,2016,22(4):290-302.
[3]陆妹,刘登礼,陆亚东,等.早产儿坏死性小肠结肠炎影响因素的回顾性分析[J].中国妇幼保健,2015,30(15):2397-2400.
[4]MORTAZ E,ADCOCK I M,TABARSI P,et al.Pattern recognitions receptors in immunodeficiency disorders[J].Eur J Pharmacol,2017,808(8):49-56.
[5]张永菊,王园园,韩一凡,等.Toll样受体1基因多态性在感染性疾病中的作用[J].实用医学杂志,2016,32(16):2758-2760.
[6]DOLASIA K,BISHT M K,PRADHAN G,et al.TLRs/NLRs:shaping the landscape of host immunity[J].Int Rev Immunol,2018,37(1):3-19.
[7]贾国泉,沈桂权,张权,等.Toll受体介导的MyD88/TRAF6/NF-κB研究[J/CD].新发传染病电子杂志,2017,2(2):104-107.
[8]JOOSTEN L A,ABDOLLAHI-ROODSAZ S,DINAREL-LO C A.et al.Toll-like receptors and chronic inflammation in rheumatic diseases:new developments[J].Nat Rev Rheumatol,2016,12(6):344-357.
[9]刘晓艳,盛德乔.Toll样受体调控适应性免疫应答的研究进展[J].细胞与分子免疫学杂志,2015,31(8):1137-1140.
[10]刘晶,白浪,苏亚茹,等.不同浓度肽聚糖对角膜上皮细胞Toll样受体(TLR)2和TLR4表达的影响[J].眼科新进展,2017,37(8):705-708.
[11]张伟,陈文利,周全,等.TOLL样受体9在动脉粥样硬化中的作用研究进展[J].激光生物学报,2016,25(4):295-301.
[12]吴迪,刘玉芬,唐敬洁.Toll样受体及其与细菌感染性疾病关系的研究进展[J].中国家禽,2016,38(17):50-52.
[13]GRAVINA H D,GOES A M,MURTA S M,et al.MyD88 Adapter-like(Mal)/TIRAP Is Required for Cytokine Production by Splenic Ly6CloTLR2hi but Not by Ly6ChiTLR2hi Monocytes during Trypanosoma cruzi Infection[J].J Biol Chem,2016,291(45):23832-23841.
[14]白琳,杨雨欣,万巧凤,等.黄芩苷经TLR2/NF-κB途径减轻类风湿关节炎大鼠滑膜炎[J].中国药理学通报,2017,33(11):1569-1573.
[15]李彩平,苏坤雄,张红,等.新生儿细菌和病毒感染T淋巴细胞亚群及细胞因子表达观察[J].临床肺科杂志,2016,21(2):280-282.
[16]洪燕燕,杨芝红.流式细胞术检测肺泡灌洗液T淋巴细胞亚群的诊断价值[J].检验医学与临床,2017,14(14):2051-2053.
[2]KAN B,RAZZAGHIAN H R,LAVOIE P M.An immunological perspective on neonatal sepsis[J].Trends Mol Med,2016,22(4):290-302.
[3]陆妹,刘登礼,陆亚东,等.早产儿坏死性小肠结肠炎影响因素的回顾性分析[J].中国妇幼保健,2015,30(15):2397-2400.
[4]MORTAZ E,ADCOCK I M,TABARSI P,et al.Pattern recognitions receptors in immunodeficiency disorders[J].Eur J Pharmacol,2017,808(8):49-56.
[5]张永菊,王园园,韩一凡,等.Toll样受体1基因多态性在感染性疾病中的作用[J].实用医学杂志,2016,32(16):2758-2760.
[6]DOLASIA K,BISHT M K,PRADHAN G,et al.TLRs/NLRs:shaping the landscape of host immunity[J].Int Rev Immunol,2018,37(1):3-19.
[7]贾国泉,沈桂权,张权,等.Toll受体介导的MyD88/TRAF6/NF-κB研究[J/CD].新发传染病电子杂志,2017,2(2):104-107.
[8]JOOSTEN L A,ABDOLLAHI-ROODSAZ S,DINAREL-LO C A.et al.Toll-like receptors and chronic inflammation in rheumatic diseases:new developments[J].Nat Rev Rheumatol,2016,12(6):344-357.
[9]刘晓艳,盛德乔.Toll样受体调控适应性免疫应答的研究进展[J].细胞与分子免疫学杂志,2015,31(8):1137-1140.
[10]刘晶,白浪,苏亚茹,等.不同浓度肽聚糖对角膜上皮细胞Toll样受体(TLR)2和TLR4表达的影响[J].眼科新进展,2017,37(8):705-708.
[11]张伟,陈文利,周全,等.TOLL样受体9在动脉粥样硬化中的作用研究进展[J].激光生物学报,2016,25(4):295-301.
[12]吴迪,刘玉芬,唐敬洁.Toll样受体及其与细菌感染性疾病关系的研究进展[J].中国家禽,2016,38(17):50-52.
[13]GRAVINA H D,GOES A M,MURTA S M,et al.MyD88 Adapter-like(Mal)/TIRAP Is Required for Cytokine Production by Splenic Ly6CloTLR2hi but Not by Ly6ChiTLR2hi Monocytes during Trypanosoma cruzi Infection[J].J Biol Chem,2016,291(45):23832-23841.
[14]白琳,杨雨欣,万巧凤,等.黄芩苷经TLR2/NF-κB途径减轻类风湿关节炎大鼠滑膜炎[J].中国药理学通报,2017,33(11):1569-1573.
[15]李彩平,苏坤雄,张红,等.新生儿细菌和病毒感染T淋巴细胞亚群及细胞因子表达观察[J].临床肺科杂志,2016,21(2):280-282.
[16]洪燕燕,杨芝红.流式细胞术检测肺泡灌洗液T淋巴细胞亚群的诊断价值[J].检验医学与临床,2017,14(14):2051-2053.