本文關(guān)鍵詞： TGF-β siRNA 乳腺上皮細(xì)胞 IL- TNF-α　出處：《中國預(yù)防獸醫(yī)學(xué)報》2017年07期 　論文類型：期刊論文

【摘要】：為研究轉(zhuǎn)化生長因子β1(TGF-β1)對脂多糖(LPS)刺激小鼠乳腺上皮細(xì)胞(MECs)分泌炎癥因子的影響,本實(shí)驗(yàn)利用si RNA轉(zhuǎn)染技術(shù)干擾MECs TGF-β1基因,采用熒光定量RT-PCR方法篩選最佳轉(zhuǎn)染條件,并檢測其干擾后LPS介導(dǎo)其下游信號smad3 m RNA的表達(dá)量,采用ELISA法檢測其下游信號smad3蛋白含量和LPS介導(dǎo)細(xì)胞分泌前炎癥細(xì)胞因子TNF-α和IL-6的含量。結(jié)果顯示,選用50 n M濃度的TGF-β1-mus-1212片段轉(zhuǎn)染組能夠極顯著抑制TGF-β1的m RNA表達(dá)(p0.01);TGF-β1被干擾后,LPS刺激其下游smad3 m RNA的表達(dá)量與對照組相比呈顯著性降低(p0.05),smad3蛋白含量與對照組相比均極顯著性降低(p0.01);細(xì)胞分泌TNF-α的量與對照組相比顯著降低(p0.05),分泌IL-6的量各組無變化(p0.05)。本實(shí)驗(yàn)結(jié)果表明干擾TGF-β1能降低LPS誘導(dǎo)乳腺上皮細(xì)胞TNF-α的分泌量,因此TGF-β1可作為靶基因深入研究乳腺上皮細(xì)胞的免疫功能,本研究為乳腺炎癥、腫瘤等乳腺相關(guān)疾病的治療提供新的思路及實(shí)驗(yàn)數(shù)據(jù),為新藥的開發(fā)和應(yīng)用提供理論依據(jù)。
[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- 偽 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- 偽 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- 偽 in breast epithelial cells induced by LPS, and the secretion of TNF- 偽 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.
【作者單位】： 貴州大學(xué)貴州省生化工程中心;貴州省畜牧獸醫(yī)研究所;貴州大學(xué)藥學(xué)院;
【基金】：國家自然科學(xué)基金項(xiàng)目(31260618、31470128) 貴大校創(chuàng)字(2016006) 黔科合LH字[2013]6014號
【分類號】：S852.3

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