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纳米脂质体体外细胞毒性作用及机制研究

In vitro cytotoxicity of HRZ(isoniazid + rifampin + pyrazinamide)/transforming growth factor β1 siRNA nanoliposomes and the underlying mechanism

目的研究异烟肼、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纳米脂质体对体外人巨噬细胞的细胞毒性作用及机制。方法利用该医院自行研制的纳米脂质体,以体外培养人巨噬细胞为研究对象,采用四唑盐(MTT)比色试验检测细胞增殖,流式细胞术检测细胞凋亡和细胞周期,电镜观察细胞自噬,反转录-PCR(RTPCR)和Western-blot检测靶基因TGF-β1沉默表达情况。结果 HRZ三联抗结核药/TGF-β1 siRNA纳米脂质体(HRZ/TGF-β1si RNA纳米脂质体)在一定浓度范围内抑制巨噬细胞增殖,使细胞周期被捕获于G2期,HRZ/TGF-β1 siRNA纳米脂质体显著抑制巨噬细胞靶基因TGF-β1的表达。结论该自行研制的HRZ/TGF-β1 siRNA纳米脂质体具有明显的靶基因沉默作用,细胞毒性在合格范围内,是一种有应用潜力的生物材料。

Objective To study the in vitro cytotoxicity of HRZ(isoniazid+rifampin+pyrazinamide)/transforming growth factor(TGF)β1 siRNA nanoliposomes on human macrophages and the underlying mechanism.Methods Self-made nanoliposomes were used to study with the cultured human macrophages in vitro.MTT assay was used to detect cell proliferation.Flow cytometry was used to analyze apoptosis and cell cycle.Electron microscopy was used to observe autophagy.RT-PCR and Western blot were employed to analyze the silenced expression of target gene TGF-β1.Results HRZ/TGF-β1 siRNA nanoliposomes(triple liposome)inhibited macrophage proliferation within certain range of concentration,and cell cycle was captured in G2 phase.The HRZ/TGF-β1 SiRNA nanoliposomes could significantly inhibit the expression of target gene TGF-β1 in human macrophages.Conclusions The self-made triple liposome has evident effect in silencing the target gene.It is a promising biomaterial,which meets the required specifications in terms of cytotoxicity.