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目的研究肺炎克雷伯菌生物膜对小鼠腹腔巨噬细胞TLRs受体表达的影响,探索机体抗生物膜(biofilm,BF)感染免疫的特点。方法将雄性昆明种小鼠40只随机分成2组,一组腹腔植入体外形成肺炎克雷伯菌生物膜的硅胶片,建立留置性医疗装置BF感染模型实验组,另一组植入与实验组同等量的浮游菌作为对照组。实时定量PCR分析2组巨噬细胞TLRs mRNA的表达水平,流式细胞仪检测分析蛋白的表达水平。结果实验生物膜组巨噬细胞TLR2、TLR4 mRNA相对表达量是对照浮游菌组的0.23和0.24倍;实验组TLR2、TLR4蛋白表达率分别是(23.27±2.73)%和(15.83±2.04)%,明显低于对照组的(33.42±3.72)%、(21.75±1.25)%(P<0.05)。结论与浮游菌相比,BF能下调小鼠腹腔巨噬细胞TLR2、TLR4表达,从而影响机体的免疫功能,这可能是BF相对浮游菌更容易逃脱机体免疫防御系统、引起慢性感染的机制之一。
Objective To study the effect of Klebsiella pneumoniae biofilm on TLRs expression in murine peritoneal macrophages and to explore the characteristics of biofilm (BF) infection. Methods Forty male Kunming mice were randomly divided into two groups. One group was implanted with silica gel sheets of Klebsiella pneumoniae biofilm in vitro, and the experimental group with BF infection model of indwelling medical device was established. The other group was implanted with experimental Groups of the same amount of planktonic bacteria as a control group. Real-time quantitative PCR was used to analyze the mRNA expression levels of TLRs in macrophages. The expression of TLRs was detected by flow cytometry. Results The relative expression levels of TLR2 and TLR4 mRNA in macrophages of experimental biofilm group were 0.23 and 0.24 times higher than those of control group. The expression rates of TLR2 and TLR4 in experimental group were (23.27 ± 2.73)% and (15.83 ± 2.04)%, respectively (33.42 ± 3.72)%, (21.75 ± 1.25)% (P <0.05) in the control group. Conclusions BF can down-regulate the expression of TLR2 and TLR4 in peritoneal macrophages compared with the planktonic bacteria and thus affect the immune function of the organism, which may be one of the mechanisms by which BF is more likely to escape the body’s immune defense system and cause chronic infection .