鄢雯 李囡 刘立新 夏山 刘扬.SCARF1 在人THP-1 单核源性巨噬细胞抗烟曲霉菌刺激中的作用[J].,2016,16(25):4815-4818 |
SCARF1 在人THP-1 单核源性巨噬细胞抗烟曲霉菌刺激中的作用 |
The Role of SCARF1 in THP1 Derived Macrophage Defense against Asperillus Fumigatus |
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DOI: |
中文关键词: SCARF1 烟曲霉菌 THP-1 细胞 NF-κB |
英文关键词: SCARF1 THP1 cells NF-κB |
基金项目:北京高等学校青年英才项目(YETP1679);首都医科大学自然科学基金项目(2014QD03) |
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中文摘要: |
目的:运用干扰腺病毒沉默THP1 细胞中SCARF1 基因研究其在体外抗烟曲霉中的作用。方法:用灭活的烟曲霉分生孢子
(1× 105 CFU / mL)于不同时间点处理THP-1 细胞,RT-PCR 分别检测SCARF1 和TNF-琢mRNA 的表达;将Ad-siRNA-SCARF1
转导细胞24 h后给予烟曲霉孢子刺激24 h,通过RT-PCR 法检测细胞中TNF-alpha mRNA表达,Western blot 法检测细胞中SCARF1
表达以及NF-kapaB 通路相关信号分子的活性。结果:RT-PCR 证实烟曲霉孢子刺激能时间依赖性增强THP1 细胞中SCARF1 和
TNF-α表达;Western法证实与Ad-GFP 组比较Ad-siRNA-SCARF1 组SCARF1 的表达量显著降低(P<0.05),沉默效率为71 %;与
Ad-GFP 组比较,Ad-GFP+Af组NF-κB 亚单位p65 的磷酸化水平明显升高(P<0.05),在Ad-siRNA-SCARF1+Af组,磷酸化p65 的
产生明显减少,SCARF1 沉默后细胞因子TNF -α的分泌明显减少。结论:烟曲霉孢子刺激能诱导巨噬细胞SCARF1 的表达增加,
诱导信号分子NF-κB 的活化,导致相应的细胞因子分泌增加,从而在巨噬细胞抗烟曲霉中发挥作用。 |
英文摘要: |
Objective:To study the role of SCARF1 against Aspergillus fumigatus in THP1 cells through knockdown of SCARF1
by adenoviruses expressing shRNA against SCARF1.Methods:The expression of SCARF1 and TNF-αgene of THP1 cells were measured
by RT-PCR after treatment with 1× 105 CFU / mL inactivated Aspergillus fumigatus spores for different time lengths. After knockdown
of SCARF1 expression with adenoviruses expressing shRNA against SCARF1, macrophages were stimulated for 24 h by Aspergillus fumigatus
conidia. The expression levels of TNF-α mRNA, SCARF1 protein and signal molecules of NF-κB pathway were examined
by RT-PCR and Western blot.Results:Aspegillus fumigatus spores stimulation can increase the expression of SCARF1 and
TNF-αin THP1 cells time-dependently. Silencing efficiency of Ad-siRNA-SCARF1 was up to 71 % in macrophages (P<0.05). Compared
with the Ad-GFP group, -p65 phosphorylation was increased significantly in Ad-GFP+Af group (P<0.05); Compared with
Ad-GFP+Af group, the p65 phosphorylation and secretion of cytokine TNF-α were lowered significantly in SCARF1 (RNAi)+Af group.Conclusion:SCARF1 in macrophages was activated by Aspergillus fumigatus conidia and exerted the effect of anti-Asp而gillus fumigatus spores
stimulation through stimulating NF-κB signaling pathway and releasing pro-inflammatory cytokines. |
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