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目的 本研究通过对新生儿中性粒细胞吞噬细菌后所产生的活性氧自由基之一的超氧阴离子进行检测,以探讨新生儿中性粒细胞功能低下的部分原因。 方法 正常足月新生儿组23例,取脐带血加以两种活菌(金黄色葡萄球菌和大肠杆菌)刺激诱发中性粒细胞活性氧代谢,用超氧阴离子特异性探针氢化溴乙非啶进行细胞内染色,通过流式细胞仪对中性粒细胞超氧阴离子产生强度进行检测;正常成人对照组15例,取外周血进行相同实验。最后采用血浆置换法探讨体液因素在中性粒细胞活性氧代谢中的作用。 结果 足月新生儿的绝大部分中性粒细胞与正常成人细胞一样随细菌的吞噬而产生活性氧自由基。金黄色葡萄球菌刺激下新生儿中性粒细胞活性氧自由基产生水平与成人基本相同;而对于大剂量大肠杆菌的刺激,其自由基产生水平较成人明显低下[细菌细胞比40∶1,平均荧光强度(MFI) 347±70和461±55, t=2.63, P<0.05; 细菌细胞比80∶1, MFI 335±66和535±76, t=4.26, P<0.01]。用成人血浆替代新生儿自身血浆后,大肠杆菌刺激下新生儿中性粒细胞超氧阴离子产生水平显著提高(MFI:707±141和461±58, t=4.91, P<0.01)。 结论 全血条件下足月新生儿中性粒细胞活性氧自由基产生水平与所吞噬细菌的种类和数量有关,在大剂量大肠杆菌的刺激下活?
Aims This study examined the neonatal neutrophil phagocytic bacteria produced by one of the active oxygen free radicals after the superoxide anion in order to explore neonatal neutrophil dysfunction in part. Methods Twenty-three normal full-term newborn infants were recruited. Umbilical cord blood was used to stimulate active oxygen metabolism of neutrophils stimulated by two kinds of live bacteria (Staphylococcus aureus and Escherichia coli). The superoxide anion- Intracellular staining was performed to detect the intensity of neutrophil superoxide anion produced by flow cytometry. In the normal adult control group, 15 cases were subjected to the same experiment with peripheral blood. Finally, the use of plasma exchange method to explore the role of humoral factors in the active oxygen metabolism of neutrophils. As a result, most neutrophils of full-term newborn babies, like normal adult cells, produce reactive oxygen species when they are swallowed by bacteria. Staphylococcus aureus stimulation neonatal neutrophil reactive oxygen species production levels and adults are basically the same; for large doses of E. coli stimulation, the level of free radicals was significantly lower than adults [bacterial cells than 40: 1, the average Fluorescence intensity (MFI) 347 ± 70 and 461 ± 55, t = 2.63, P <0.05; bacterial cell ratio 80: 1, MFI 335 ± 66 and 535 ± 76, t = 4.26, P <0.01]. Neonatal neonatal neutrophil superoxide anion production was significantly increased after stimulation with E. coli (MFI: 707 ± 141 and 461 ± 58, t = 4.91, P <0.01) after replacement of neonatal plasma with adult plasma. Conclusion The production of reactive oxygen species of full-term neonatal neutrophils in whole blood is related to the type and quantity of phagocytosed bacteria. Under the stimulation of high-dose Escherichia coli,