结核感染患者外周血中性粒细胞自噬水平的研究

Study on the level of neutrophil autophagy in peripheral blood of patients with tuberculosis infection

目的探讨结核感染患者外周血中性粒细胞自噬水平。方法根据结核病诊断标准,将患者分为健康对照组(16例)和活动性结核组(24例),收集患者外周血,采用流式细胞技术分别检测健康对照组和活动性结核组中性粒细胞和单个核细胞的自噬状态。分离外周血中性粒细胞,采用Trizol法提取总RNA,经逆转录实时荧光定量PCR,利用2-△△Ct计算Beclin1表达的相对定量。结果活动性结核组与健康对照组年龄差异无统计学意义(P=0.165 5,P>0.05);活动性结核组男性比例(79.2%)明显高于健康对照组(37.5%)。活动性结核组中性粒细胞自噬水平显著低于健康对照组(P=0.013 8,P<0.05),而单个核细胞两组差异无统计学意义(P=0.784 2,P>0.05);活动性结核组中性粒细胞Beclin1△Ct=-1.254±0.40明显低于健康对照组△Ct=-0.105±0.48,差异有统计学意义(P<0.001)。结论结核感染患者外周血中性粒细胞的自噬水平降低,且自噬相关基因Beclin1mRNA的表达下调,中性粒细胞的自噬水平可能与结核病的发生、发展密切相关。

Objective To investigate the autophagy level of peripheral blood neutrophils in patients with tuberculosis infection.Methods According to the standard for tuberculosis diagnosis,the subjects were divided into normal control group(16 cases)and active tuberculosis group(24 cases),and peripheral blood was collected.The autophagy of neutrophils and mononuclear cells of the two groups were detected by flow cytometry:The total RNA of neutrophils was extracted by Trizol and the relative quantification of Beclin1 expression was calculated by using 2-^△△Ct in real-time quantitative PCR.Results There was no significant difference in age between the active tuberculosis and normal control group(P=0.165 5,P>0.05);the proportion of male in active tuberculosis group(79.2%)was significantly higher than that of normal control group(37.5%),and the difference was statistically significant(P=0.031 5,P<0.05).The level of neutrophil autophagy in the active tuberculosis group was significantly lower than that of the normal control group(P=0.013 8,P<0.05),and there was no difference of mononuclear cells between the two groups(P=0.784 2,P>0.05);active tuberculosis group the mRNA expression of Beclin1△Ct=-1.254±0.40 was lower than that of the normal control group△Ct=-0.10±0.48,the difference was statistically significant(P<0.001).Conclusion The incidence of active tuberculosis in male is higher than that of female;the autophagy level of neutrophils in peripheral blood of patients with tuberculosis infection decreased,and the decline of autophagy-related gene Beclin1 mRNA expression may be related to the occurrence and development of tuberculosis.