微小RNA-146a在新生隐球菌、格特隐球菌诱导THP-1细胞炎性反应中的机制研究

发布时间：2018-08-08 11:01

【摘要】：目的观察并分析微小RNA-146a(miR-146a)在新生隐球菌(标准株WM148)、格特隐球菌(标准株R265)刺激人单核巨噬细胞系(THP-1细胞)中表达的差异,探讨miR-146a在隐球菌所致隐球菌脑膜炎中的调控作用及机制。方法体外培养的人单核巨噬细胞系THP-1细胞分成新生隐球菌诱导组、格特隐球菌诱导组,按灭活菌数∶THP-1=5∶1的比例与THP-1细胞共孵育0、3、6、9和12 h后离心收集上清液和细胞沉淀。采用实时荧光定量PCR(q RT-PCR)技术检测各组细胞miR-146a的表达情况,酶联免疫吸附试验(ELISA)法检测各组上清液中肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)的释放量。结果新生隐球菌诱导组细胞中miR-146a表达量与0 h相比明显升高(P0.01),且在3 h达到峰值,随后呈下降趋势;上清液中TNF-α的表达量在诱导后12 h达到最高值,IL-6的表达各时间点没有明显变化。格特隐球菌诱导后,miR-146a的表达逐渐升高,12 h达到最高值,但3 h、6 h点变化与0 h比较差异并无统计学意义;上清液中TNF-α的表达12 h到达峰值;IL-6的表达逐渐增加,12 h达到最高值,变化无统计学意义。结论新生隐球菌、格特隐球菌刺激THP-1细胞炎性反应后,miR-146a表达随时间延长呈现不同的规律,TNF-α、IL-6的动态变化表现不同的特点。研究表明新生隐球菌、格特隐球菌诱导THP-1炎性反应可能存在不同的调控机制。
[Abstract]:Objective to observe and analyze the difference of the expression of minimal RNA-146a (miR-146a) in Cryptococcus neoformans (WM148) and Cryptococcus Gert (R265) stimulated human mononuclear macrophage cell line (THP-1), and to explore the regulatory role and mechanism of miR-146a in Cryptococcus neoformans meningococcal meningitis. Methods Human mononuclear macrophage THP-1 cells were divided into two groups: Cryptococcus neoformans induction group and Gert Cryptococcus induction group. The supernatants and cell precipitates were collected by centrifugation after incubation with THP-1 cells in proportion of 1: THP-1 to 5: 1 for 12 h. The expression of miR-146a was detected by real-time fluorescence quantitative PCR (q RT-PCR, and the release of tumor necrosis factor- 伪 (TNF- 伪) and interleukin-6 (IL-6) in supernatant of each group was detected by enzyme-linked immunosorbent assay (ELISA). Results the expression of miR-146a in Cryptococcus neoformans group was significantly higher than that in 0 h group (P0.01), and reached its peak at 3 h, and then decreased. The expression of TNF- 伪 in supernatant did not change significantly at 12 h after induction. The expression of miR-146a increased gradually and reached the highest level at 12 h after induction, but there was no significant difference between 3 h and 0 h, and the expression of TNF- 伪 in supernatant reached the peak at 12 h and reached the peak value at 12 h. The changes were not statistically significant. Conclusion the expression of miR-146a in Cryptococcus neoformans and Cryptococcus geterii stimulated inflammatory response of THP-1 cells showed different regularity with time. The dynamic changes of IL-6 in TNF- 伪 were different. It has been suggested that Cryptococcus neoformans and Cryptococcus gertsii may have different regulatory mechanisms for THP-1 inflammatory response.
【作者单位】：第二军医大学附属长征医院皮肤科;南京金陵医院皮肤科;
【基金】：国家重点专科建设项目
【分类号】：R519.4

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