siRNA干扰TGF-β1基因对LPS诱导小鼠乳腺上皮细胞IL-6和TNF-α分泌的影响

发布时间：2018-02-09 02:27

本文关键词： TGF-β siRNA 乳腺上皮细胞 IL- TNF-α　出处：《中国预防兽医学报》2017年07期 　论文类型：期刊论文

【摘要】：为研究转化生长因子β1(TGF-β1)对脂多糖(LPS)刺激小鼠乳腺上皮细胞(MECs)分泌炎症因子的影响,本实验利用si RNA转染技术干扰MECs TGF-β1基因,采用荧光定量RT-PCR方法筛选最佳转染条件,并检测其干扰后LPS介导其下游信号smad3 m RNA的表达量,采用ELISA法检测其下游信号smad3蛋白含量和LPS介导细胞分泌前炎症细胞因子TNF-α和IL-6的含量。结果显示,选用50 n M浓度的TGF-β1-mus-1212片段转染组能够极显著抑制TGF-β1的m RNA表达(p0.01);TGF-β1被干扰后,LPS刺激其下游smad3 m RNA的表达量与对照组相比呈显著性降低(p0.05),smad3蛋白含量与对照组相比均极显著性降低(p0.01);细胞分泌TNF-α的量与对照组相比显著降低(p0.05),分泌IL-6的量各组无变化(p0.05)。本实验结果表明干扰TGF-β1能降低LPS诱导乳腺上皮细胞TNF-α的分泌量,因此TGF-β1可作为靶基因深入研究乳腺上皮细胞的免疫功能,本研究为乳腺炎症、肿瘤等乳腺相关疾病的治疗提供新的思路及实验数据,为新药的开发和应用提供理论依据。
[Abstract]:In order to study the effect of transforming growth factor 尾 1 (TGF- 尾 1) on the secretion of inflammatory cytokines in murine mammary epithelial cells stimulated by lipopolysaccharide (LPS), we used si RNA transfection technique to interfere with MECs TGF- 尾 1 gene, and used fluorescence quantitative RT-PCR to screen the best transfection conditions. The expression of downstream signal smad3 m RNA mediated by LPS was detected. The content of downstream signal smad3 protein and LPS mediated preinflammatory cytokines TNF- 伪 and IL-6 were detected by ELISA assay. The expression of m RNA of TGF- 尾 _ 1-mus-1212 fragment transfected with 50 mm concentration of TGF- 尾 _ 1 was significantly inhibited in the control group. After the interference, the expression of smad3 m RNA in the downstream of TGF- 尾 _ 1 was significantly decreased compared with that in the control group, and the protein level of Smad3 was significantly lower than that of the control group. The amount of TNF- 伪 secreted by the cells was significantly lower than that of the control group (P 0.05). The results showed that interfering with TGF- 尾 1 could decrease the secretion of TNF- 伪 in breast epithelial cells induced by LPS, and the secretion of TNF- 伪 in the breast epithelial cells induced by TGF- 尾 1 could be decreased by interfering with TGF- 尾 1. Therefore, TGF- 尾 1 can be used as a target gene to further study the immune function of breast epithelial cells. This study provides new ideas and experimental data for the treatment of mammary gland related diseases such as breast inflammation and tumor, and provides theoretical basis for the development and application of new drugs.
【作者单位】：贵州大学贵州省生化工程中心;贵州省畜牧兽医研究所;贵州大学药学院;
【基金】：国家自然科学基金项目(31260618、31470128) 贵大校创字(2016006) 黔科合LH字[2013]6014号
【分类号】：S852.3

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