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目的本研究旨在评估肺炎链球菌性急性中耳炎(S.pneumoniae, S.pn;Acute Otitis Media, S.pn AOM)中一氧化氮(NO)通过促进糖酵解影响细菌清除和炎症应答的作用。方法听泡穿刺接种肺炎链球菌血清型19F建立AOM小鼠模型,荧光定量PCR技术检测中耳灌洗液中中性粒细胞糖酵解相关基因的表达变化。诱导型一氧化氮合酶(iNOS)抑制剂1400W及糖酵解抑制剂2-脱氧葡萄糖(2-deoxy-D-glucose,2DG)分别处理后,观察糖酵解代谢变化、免疫应答及对细菌清除的影响。结果 S.pn AOM时,中耳灌洗液中中性粒细胞呈现糖酵解关键酶、葡萄糖转运体及乳酸分泌转运体等表达上调并伴有葡萄糖消耗增加和乳酸生成增多的免疫代谢特征(P<0.05),同时,iNOS表达显著上调,NO生成增加(P<0.05)。糖酵解促进炎症细胞渗出、细胞因子分泌和S.pn清除(P<0.05)。iNOS抑制剂处理后,NO生成显著减少,糖酵解受抑,炎症反应减轻,细菌清除受损(P<0.05)。结论 S.pn AOM时,中耳腔中中性粒细胞糖酵解增强,释放NO促进糖酵解,进而增强中耳炎症应答和S.pn清除。
Abstract：Objective The aim of this study is to evaluate the role of NO in inflammatory responses and bacterial clearance by regulating glycolysis in a mouse model of acute otitis media(AOM). Methods The AOM model was established by transbullar puncture inoculation of Streptococcus pneumoniae(S. pn) serotype 19 F. Expression levels of glycolytic related genes in neutrophils in middle ear lavage were evaluated by fluorescent quantitative PCR. Changes in glycolytic metabolism, immune responses and bacterial clearance were evaluated after treatment with 1400 W(an iNOS inhibitor) and 2-deoxy-D-glucose(2 DG, a glycolysis inhibitor), respectively. Results During S.pn AOM, expression of key glycolytic enzymes Hk3, Pfkfb3, Pkm2, glucose transporter glut1 and lactate transporter Mct4 in neutrophils in middle ear lavage, as well as glucose consumption and lactic acid production, significantly increased, suggesting enhancement of glycolysis. Meanwhile, expression of iNOS and production of NO also increased significantly(P<0.05). Moreover,exudation of inflammatory cells, secretion of cytokines and bacterial clearance were promoted by glycolysis(P<0.05).After treatment with 1400 W, production of NO and levels of glycolysis and inflammatory response all decreased, accompanied by inhibition of bacterial clearance(P<0.05).Conclusion During S.pn AOM, neutrophils recruited into the middle ear cavity release NO to promote glycolysis, thus enhancing inflammatory responses and bacterial clearance.
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基本信息:
中图分类号:R764.21
引用信息:
[1]艾容霜,范芳梅,马毓蓉,等.NO促进糖酵解增强急性中耳炎的炎症应答和细菌清除[J].中华耳科学杂志,2021,19(03):480-485.
基金信息:
Chongqing Research Program of Basic Research and Frontier Technology(cstc2018jcyjAX0257)
2021-06-11
2021-06-11