微小RNA-146a-5p在新生隐球菌、脂多糖诱导THP-1细胞中表达的对比分析

发布时间：2018-03-17 09:42

本文选题：微小RNA-a-p　切入点：新生隐球菌　出处：《中国感染与化疗杂志》2015年06期 　论文类型：期刊论文

【摘要】：目的为探讨微小RNA-146a-5p(miR-146a-5p)在新生隐球菌所致隐球菌性脑膜炎中的调控作用及机制奠定基础。方法把体外培养的人单核细胞系THP-1细胞分成新生隐球菌刺激组和脂多糖(LPS)刺激组,分别用与细胞数量相同的灭活新生隐球菌(WM148)和浓度为1 mg/L的LPS刺激THP-1细胞,分别于0、3、6、9、12 h收集上清液和细胞。采用酶联免疫吸附试验(ELISA)检测细胞上清液中肿瘤坏死因子(TNF)-α、白细胞介素(IL)-6的表达水平,实时荧光定量聚合酶链反应(Real-time PCR)检测细胞miR-146a-5p的表达情况。结果新生隐球菌诱导后,细胞中miR-146a-5p的表达对比对照组(0h)明显升高(P0.05),3 h达到最高值,随后逐渐下降;上清液中TNF-α的表达在诱导后12 h达到最高值,IL-6的表达各时间点变化不明显。LPS诱导后,miR-146a-5p的表达逐渐升高,12 h达到最高值;上清液中TNF-α的表达3 h后一直处于峰值;IL-6的表达逐渐增加,12 h达到最高值。结论新生隐球菌、LPS诱导THP-1细胞炎性反应后,miR-146a-5p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。表明miR-146a-5p参与新生隐球菌、LPS诱导THP-1炎性反应存在不同的调控机制。
[Abstract]:Objective to study the micro RNA-146a-5p (miR-146a-5p) basis function and regulatory mechanism in Cryptococcus neoformans induced cryptococcal meningitis. Methods the in vitro cultured human monocytic THP-1 cells into Cryptococcus neoformans stimulation group and lipopolysaccharide (LPS) stimulation group, respectively with the same number of cells inactivated Cryptococcus aureus (WM148) and the concentration of 1 mg/L LPS stimulation of THP-1 cells, h and 0,3,6,9,12 respectively. Cell supernatant was collected by enzyme-linked immunosorbent assay (ELISA) detected in the supernatant of tumor necrosis factor (TNF) - alpha, interleukin (IL) -6 expression, real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) to detect the expression of miR-146a-5p cells. The results of Cryptococcus neoformans induced expression of miR-146a-5p in cells compared with control group (0h) was significantly increased (P0.05), 3 h reached a maximum value, then decreased gradually; the supernatant Expression of TNF- in the liquid after induction of 12 h reached the highest value, the expression of IL-6 at each time point did not change significantly after induction of.LPS miR-146a-5p expression gradually increased, H 12 has reached the highest value; at the peak 3 h after the expression of TNF- in the supernatant; the expression of IL-6 increased gradually, reached the highest value of 12 h conclusion Cryptococcus neoformans, LPS THP-1 cells induced by inflammatory reaction after miR-146a-5p expression with time show different patterns, dynamic changes of TNF- alpha, IL-6 showed different characteristics. Suggested the involvement of miR-146a-5p in Cryptococcus neoformans, LPS THP-1 induced inflammatory response in different regulatory mechanisms.

【作者单位】：第二军医大学附属长征医院皮肤科;
【分类号】：R519.4

【相似文献】