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肺炎支原体脂质相关膜蛋白刺激 THP-1细胞表达血红素氧合酶-1

The lipid-associated membrane proteins derived from Mycoplasma pneumoniae strains induced the expression of HO-1 in THP-1 cells

摘要目的:研究肺炎支原体( Mycoplasma pneumonia, Mp)脂质相关膜蛋白( lipid-associated membrane proteins , LAMPs)对人单核细胞系THP-1表达血红素氧合酶-1(heme oxygenase 1,HO-1)的影响。方法体外培养THP-1细胞,根据实验目的加入不同浓度LAMPs作用不同时间,并设阳性和阴性对照。收集各处理组细胞,LDH漏出实验检测LAMPs对THP-1细胞的毒性,实时荧光反转录PCR检测 HO-1 mRNA 表达, Western blot 检测 HO-1蛋白表达,比色法检测 HO-1酶活性。结果LAMPs浓度为10μg/ml时,LDH漏出率明显增高。 LAMPs刺激THP-1细胞9 h后HO-1 mRNA表达水平最高;HO-1蛋白表达水平与LAMPs刺激呈剂量依赖性和时间依赖性,其中5.0μg/ml LAMPs刺激表达的HO-1蛋白水平最高;LAMPs刺激12 h 后的HO-1蛋白浓度最高。 LAMPs刺激THP-1细胞后,随着HO-1表达水平的增高,其酶活性亦显著增强。结论 Mp LAMPs可诱导THP-1细胞表达HO-1,并可上调其酶活性。

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abstractsObjective To investigate the effects of lipid-associated membrane proteins ( LAMPs) derived from Mycoplasma pneumoniae ( M.pneumoniae) strains on the expression of heme oxygenase 1 ( HO-1) in a human monocyte cell line (THP-1).Methods THP-1 cells were in vitro cultured with different concentrations of LAMPs for different times.The cytotoxicity of LAMPs to THP-1 cells was analyzed by using lactate dehydrogenase ( LDH) releasing test.The expression of HO-1 at protein and mRNA levels were de-tected by Western blot and real-time RT-PCR, respectively.The enzymatic activity of HO-1 protein was ex-amined by colorimetric assay.THP-1 cells stimulated with PBS and LPS were set up as the negative and pos-itive controls, respectively.Results A significantly enhanced LDH releasing rate was observed in THP-1 cells treated with 10 μg/ml of LAMPs.The expression of HO-1 at protein and mRNA levels in THP-1 cells were induced by LAMPs in a dose-dependent and time-dependent manner.The highest level of HO-1 protein was detected in THP-1 cells treated with 5.0 μg/ml of LAMPs.The transcriptional levels of HO-1 induced by LAMPs were significantly elevated at 3 h, peaked at 9 h and were decreased at 12 h.The expression of HO-1 protein in THP-1 cells was enhanced after 8 h of treatment with LAMPs and a significant decrease was observed at 20 h after reaching peaks at 12 h and 16 h.The activity of HO-1 protein was significantly en-hanced along with the increased expression of HO-1 protein.Conclusion The LAMPs derived from M.pneumoniae strains induced the expression of HO-1 at mRNA and protein levels.Moreover, the enzyme activity of HO-1 protein was enhanced in LAMPs treated THP-1 cells.

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栏目名称 基础免疫学
DOI 10.3760/cma.j.issn.0254-5101.2015.02.006
发布时间 2015-04-23
基金项目
国家自然科学基金(81441065) 湖南省自然科学基金(14JJ2090) 湖南省高校创新平台开放基金(14K086) 特殊病原体防治湖南省高校重点实验室 湖南省科技厅重点实验室基金
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