牛宁奎,吴龙云,何进文,王立楠,杨宗强,师志云,施建党,王自立,丁惠强.HRZ三联抗结核药/TGF-β1 siRNA纳米脂质体对体外BCG感染的人巨噬细胞中Ag85A及TGF-β1表达的影响[J].中国脊柱脊髓杂志,2018,(10):918-924. |
HRZ三联抗结核药/TGF-β1 siRNA纳米脂质体对体外BCG感染的人巨噬细胞中Ag85A及TGF-β1表达的影响 |
中文关键词: 异烟肼 利福平 吡嗪酰胺 三联抗结核药/表皮转化生长因子-β1 siRNA纳米脂质体 巨噬细胞 抗结核 靶向基因沉默 |
中文摘要: |
【摘要】 目的:探讨异烟肼、利福平、吡嗪酰胺(HRZ)三联抗结核药/表皮转化生长因子(TGF)-β1 siRNA纳米脂质体对体外BCG感染的人巨噬细胞中Ag85A及TGF-β1表达的影响。方法:体外培养人单核细胞株THP-1诱导分化成巨噬细胞,分为4组,空白组为单纯巨噬细胞培养;模型组为巨噬细胞与卡介苗(bacillus calmette-guerin,BCG)以1∶5的比例共培养3h,制备BCG感染的巨噬细胞模型;对照组为BCG感染的巨噬细胞与HRZ三联抗结核药纳米脂质体共培养;实验组为BCG感染的巨噬细胞与三种不同浓度的HRZ三联抗结核药/TGF-β1 siRNA纳米脂质体(C1组为35mg/ml,C2组为40mg/ml,C3组为50mg/ml)共培养。采用RT-PCR检测各组人巨噬细胞中Ag85A及TGF-β1的mRNA表达量,Western-blot方法检测Ag85A及TGF-β1的蛋白表达量。结果:与空白组相比,模型组Ag85A和TGF-β1的mRNA及蛋白量表达明显上调(P<0.05);对照组中Ag85A的mRNA及蛋白量和TGF-β1的mRNA水平与模型组相比明显下调(P<0.05);实验组3种不同浓度(C1组、C2组、C3组)HRZ三联抗结核药/TGF-β1 siRNA纳米脂质体处理后,Ag85A和TGF-β1的mRNA及蛋白表达量与模型组和对照组相比进一步下调(P<0.05),随着纳米脂质体浓度的增加,Ag85A和TGF-β1 mRNA及蛋白表达量有明显下降趋势,C1组与C3组相比Ag85A mRNA及蛋白表达量差异有统计学意义(P<0.05);C1组与C2组、C3组相比TGF-β1 mRNA及蛋白量表达差异有统计学意义(P<0.05);C2组与C3组相比Ag85A mRNA和TGF-β1蛋白表达量差异有统计学意义(P<0.05)。结论:HRZ三联抗结核药/TGF-β1 siRNA纳米脂质体对体外BCG感染的人巨噬细胞具有明显的下调Ag85A基因表达作用及TGF-β1基因沉默作用。 |
Effects of HRZ triple antituberculosis drug/TGF-β1 siRNA nanoliposomes on the expression of Ag85A and TGF-β1 in human BCG-infected human macrophages |
英文关键词:Isoniazid/rifampicin/pyrazinamide(HRZ) Triple antituberculosis drug/transforming growth factor-β1 siRNA nanoliposomes Macrophage Antituberculosis Targeted gene silencing |
英文摘要: |
【Abstract】 Objectives: To investigate the effects of isoniazid/rifampicin/pyrazinamide(HRZ) triple anti-tuberculosis/epidermal transforming growth factor(TGF)-β1 siRNA nanoliposomes on the expression of Ag85A and TGF-β1 in human macrophages in vitro. Methods: The human mononuclear cell strain THP-1 was induced to differentiate into macrophages in vitro. The experiment was divided into 4 groups. The blank group was cultured as macrophage alone. In the model group, macrophage and bacillus calmette-guerin(BCG) were cocultured at the ratio of 1∶5 for 4h to prepare a BCG-infected macrophage model. In the control group, BCG-infected macrophages and HRZ triple antituberculosis nanoliposomes were cocultured. In the experimental group, BCG-infected macrophages and HRZ triple anti-tuberculosis drug/TGF-β1 siRNA nanoliposome(C1 35mg/ml, C2 40mg/ml, and C3 50mg/ml) were cococultured. The mRNA expression levels of Ag85A and TGF-β1 in human macrophages were detected by RT-PCR. The protein expression levels of Ag85A and TGF-β1 were detected by western-blot method. Results: RT-PCR and western-blot results showed that the mRNA and protein levels of Ag85A and TGF-β1 were significantly up-regulated in the model group(BCG-infected macrophage model) compared with the blank group(macrophage culture alone)(P<0.05). The mRNA and protein expressions of Ag85A and TGF-β1 in the control group(cocultured with BCG-infected macrophages and HRZ triple anti-tuberculosis nano-liposomes) were significantly down-regulated compared with the model group(P<0.05); Compared with the model group and the control group, the mRNAs and protein expressions of Ag85A and TGF-β1 treated with HRZ triple antituberculosis drugs/TGF-β1 siRNA nanoliposomes in three different concentrations(C1 group, C2 group and C3 group) in the experimental group was further down-regulated(P<0.05). With the increase of nano-liposome concentration(35mg/ml, 40mg/ml, 50mg/ml), the expressions of TGF-β1 mRNA and protein decreased significantly(the difference between C1 group and C2 group and C3 group was statistically significant, P<0.05). Conclusions: The self-developed HRZ triple anti-tuberculosis drug/TGF-β1 siRNA nano-liposome has obvious down-regulation of Ag85A gene expression and TGF-β1 gene silencing. |
投稿时间:2018-07-01 修订日期:2018-08-30 |
DOI: |
基金项目:国家自然科学基金资助项目(81501903);国家自然科学基金资助项目(81860395);宁夏自然科学基金资助项目(NZ16157);宁夏高等学校一流学科建设(宁夏医科大学国内一流建设学科临床医学)资助项目(NXYLXK2017A05);2015年宁夏留学人员资助项目;2017年宁夏医科大学青年骨干人才培育计划项目 |
|
摘要点击次数: 2802 |
全文下载次数: 2067 |
查看全文 查看/发表评论 下载PDF阅读器 |
关闭 |