【摘要】：目的觀察并分析微小RNA-146a(miR-146a)在新生隱球菌(標(biāo)準(zhǔn)株WM148)、格特隱球菌(標(biāo)準(zhǔn)株R265)刺激人單核巨噬細(xì)胞系(THP-1細(xì)胞)中表達的差異,探討miR-146a在隱球菌所致隱球菌腦膜炎中的調(diào)控作用及機制。方法體外培養(yǎng)的人單核巨噬細(xì)胞系THP-1細(xì)胞分成新生隱球菌誘導(dǎo)組、格特隱球菌誘導(dǎo)組,按滅活菌數(shù)∶THP-1=5∶1的比例與THP-1細(xì)胞共孵育0、3、6、9和12 h后離心收集上清液和細(xì)胞沉淀。采用實時熒光定量PCR(q RT-PCR)技術(shù)檢測各組細(xì)胞miR-146a的表達情況,酶聯(lián)免疫吸附試驗(ELISA)法檢測各組上清液中腫瘤壞死因子-α(TNF-α)、白細(xì)胞介素-6(IL-6)的釋放量。結(jié)果新生隱球菌誘導(dǎo)組細(xì)胞中miR-146a表達量與0 h相比明顯升高(P0.01),且在3 h達到峰值,隨后呈下降趨勢;上清液中TNF-α的表達量在誘導(dǎo)后12 h達到最高值,IL-6的表達各時間點沒有明顯變化。格特隱球菌誘導(dǎo)后,miR-146a的表達逐漸升高,12 h達到最高值,但3 h、6 h點變化與0 h比較差異并無統(tǒng)計學(xué)意義;上清液中TNF-α的表達12 h到達峰值;IL-6的表達逐漸增加,12 h達到最高值,變化無統(tǒng)計學(xué)意義。結(jié)論新生隱球菌、格特隱球菌刺激THP-1細(xì)胞炎性反應(yīng)后,miR-146a表達隨時間延長呈現(xiàn)不同的規(guī)律,TNF-α、IL-6的動態(tài)變化表現(xiàn)不同的特點。研究表明新生隱球菌、格特隱球菌誘導(dǎo)THP-1炎性反應(yīng)可能存在不同的調(diào)控機制。
[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- 偽 (TNF- 偽) 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- 偽 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- 偽 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- 偽 were different. It has been suggested that Cryptococcus neoformans and Cryptococcus gertsii may have different regulatory mechanisms for THP-1 inflammatory response.
【作者單位】： 第二軍醫(yī)大學(xué)附屬長征醫(yī)院皮膚科;南京金陵醫(yī)院皮膚科;
【基金】：國家重點專科建設(shè)項目
【分類號】：R519.4

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