脐血与成人外周血粒细胞Toll样受体表达的比较

发布时间：2018-11-17 12:50

【摘要】：目的:比较脐血与成人外周血粒细胞Toll样受体(TLRs)的表达情况,为新生儿相关疾病的临床治疗积累实验资料。方法:取脐血和成人外周血,用密度梯度离心结合红细胞裂解法分离、收集粒细胞,流式细胞术鉴定粒细胞纯度,RT-qPCR检测TLRs 10个成员的mRNA表达水平,流式细胞术测定部分TLRs的蛋白荧光强度。结果:(1)流式细胞术检测结果:采用密度梯度离心结合红细胞裂解法分离收集的脐血粒细胞CD19-CD24+为(95.66±1.73)%,CD3+为(4.27±1.22)%,成人外周血粒细胞CD19-CD24+为(95.48±2.13)%,CD3+为(4.82±1.07)%。(2)RT-qPCR结果显示:脐血粒细胞和成人外周血粒细胞TLR1(0.141±0.091 vs 0.691±0.447)、TLR2(0.388±0.337 vs 0.901±0.508)、TLR4(0.093±0.071 vs 0.254±0.147)、TLR6(0.056±0.045 vs 0.202±0.034)、TLR7(0.001±0.001 vs 0.004±0.003)和TLR8(0.046±0.040 vs 0.211±0.146)的表达差异有统计学意义(P0.01),TLR3、TLR5、TLR9和TLR10的表达差异无统计学意义(P0.05);其中TLR3、TLR7和TLR9在脐血和成人外周血粒细胞的相对表达水平均较低。(3)流式分析显示脐血与成人外周血粒细胞TLR2平均蛋白荧光强度(21.40±3.09 vs 30.50±5.69)差异有统计学意义(P0.05);而TLR4平均蛋白荧光强度差异无统计学意义(P0.05)。结论:脐血粒细胞TLR1、TLR2、TLR4和TLR6 mRNA及TLR2蛋白表达低于成人外周血粒细胞,提示新生儿粒细胞识别细菌性病原微生物感染的能力可能存在缺陷或尚未完全成熟。这是否与新生儿急性细菌性毒血症发病及病死率较高有直接联系,有待进一步研究。
[Abstract]:Objective: to compare the expression of Toll like receptor (TLRs) in umbilical cord blood and adult peripheral blood in order to accumulate experimental data for clinical treatment of neonatal diseases. Methods: umbilical cord blood and adult peripheral blood were isolated by density gradient centrifugation and erythrocyte lysis. Granulocytes were collected, granulocyte purity was identified by flow cytometry, and mRNA expression of 10 members of TLRs was detected by RT-qPCR. The fluorescence intensity of some TLRs proteins was measured by flow cytometry. Results: (1) flow cytometry: the CD19-CD24 and CD3 of cord blood granulocytes collected by density gradient centrifugation and erythrocyte lysis were (95.66 卤1.73)% and (4.27 卤1.22)%, respectively. The CD19-CD24 of adult peripheral blood granulocytes was (95.48 卤2.13)%, and that of CD3 was (4.82 卤1.07)%. (2) the results of RT-qPCR showed that the TLR1 of cord blood granulocytes and adult peripheral blood granulocytes was (0.141 卤0.091 vs) 0.691 卤0.447. TLR2 (0.388 卤0.337 vs 0.901 卤0.508), TLR4 (0.093 卤0.071 vs 0.254 卤0.147), TLR6 (0.056 卤0.045 vs 0.202 卤0.034); The expression of TLR7 (0.001 卤0.001 vs 0.004 卤0.003) and TLR8 (0.046 卤0.040 vs 0.211 卤0.146) was significantly different (P0.01), but the expression of TLR3,TLR5,TLR9 and TLR10 had no significant difference (P0.05). In which TLR3, The relative expression level of TLR7 and TLR9 in umbilical cord blood and adult peripheral blood granulocyte was lower. (3) flow cytometry showed that the average fluorescence intensity of TLR2 protein in umbilical cord blood and adult peripheral blood granulocyte was (21.40 卤3.09 vs 30.50 卤5.69). There was statistical significance (P0.05); But TLR4 average protein fluorescence intensity difference has no statistical significance (P0.05). Conclusion: the expression of TLR1,TLR2,TLR4, TLR6 mRNA and TLR2 protein in umbilical cord blood granulocyte is lower than that in adult peripheral blood granulocyte, suggesting that the ability of neonate granulocyte to recognize bacterial pathogenic microorganism infection may be deficient or immature. Whether this is directly related to the incidence and mortality of neonatal acute bacterial toxemia needs further study.
【作者单位】：暨南大学医学院田家炳医学实验中心暨南大学第一临床医院暨南大学医学院血液病研究所
【基金】：广东省产业技术研究与开发资金计划项目(No.2010B031600107) 广东省自然科学基金资助项目(No.07005966)
【分类号】：R392.1

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