NF-κB信号通路在金黄色葡萄球菌诱导奶牛乳腺成纤维细胞转分化中的作用

发布时间：2018-06-11 22:45

本文选题：金黄色葡萄球菌 + 奶牛乳腺成纤维细胞　；参考：《中国兽医学报》2017年09期

【摘要】：用热灭活金黄色葡萄球菌(0,104,105,106,107和108 CFU/mL)刺激奶牛乳腺成纤维细胞,24h后采用Real time-PCR方法检测TNF-α、IL-1β、IL-6和TGF-β1mRNA的表达量,Western blot方法检测α-SMA、collagen-I及pNF-κB p65蛋白的表达量。使用NF-κB通路抑制剂PDTC预处理细胞,再用105 CFU/mL热灭活金黄色葡萄球菌刺激细胞,24h后采用Real time-PCR方法检测TNF-α、IL-1β、IL-6和TGF-β1 mRNA的表达量,Western blot方法检测α-SMA、collagen-I及p-NF-κB p65蛋白的表达量,免疫荧光法检测α-SMA及collagen-I蛋白的表达量。结果显示,不同浓度热灭活金黄色葡萄球菌处理细胞的TGF-β1mRNA的表达量显著升高(P0.05),其中以105 CFU/mL刺激组的TGF-β1mRNA的表达量最高。随着热灭活金黄色葡萄球菌浓度的升高,p-NF-κB p65蛋白的表达量逐渐升高(P0.01),TNF-α、IL-1β、IL-6mRNA的表达量也逐渐升高(P0.01)。不同浓度灭活的金黄色葡萄球菌处理细胞的α-SMA及collagen-I蛋白的表达量均能显著升高(P0.05),其中以105 CFU/mL刺激组的α-SMA及collagen-I蛋白的表达量最高。抑制剂PDTC能够显著抑制TNF-α、IL-1β、IL-6和TGF-β1 mRNA的表达量(P0.05),PDTC能够显著抑制α-SMA、collagen-I及p-NF-κB p65蛋白的表达量(P0.05)。结果提示,NF-κB信号通路在金黄色葡萄球菌诱导奶牛乳腺成纤维细胞转分化为肌成纤维细胞过程中发挥重要作用,本研究为揭示金黄色葡萄球菌性乳腺炎发生硬化的机制奠定基础。
[Abstract]:The expression of TNF- 伪, IL-1 尾, IL-6 and TGF- 尾 1 mRNA in bovine mammary fibroblasts were detected by Real time-PCR method. The expressions of 伪 -SMA-collagen-I and pNF- 魏 B p65 protein were detected by Western blot method after stimulation with thermally inactivated Staphylococcus aureus 0104105106107 and 108CFU / mL. The cells were pretreated with NF- 魏 B pathway inhibitor PDTC, and then incubated with 105 CFU / mL heat inactivated Staphylococcus aureus cells for 24 hours. The expression of TNF- 伪 -IL-1 尾 -IL-6 and TGF- 尾 _ 1 mRNA was detected by Real time-PCR method. The expression of 伪 -SMA-collagen-I and p-NF- 魏 B p65 protein was detected by Western blot. The expression of 伪 -SMA and collagen-I protein was detected by immunofluorescence. The results showed that the expression of TGF- 尾 1 mRNA was significantly increased in Staphylococcus aureus cells treated with different concentrations of heat inactivated Staphylococcus aureus, and the expression of TGF- 尾 1 mRNA was the highest in the 105 CFU / mL stimulated group. With the increase of heat inactivated Staphylococcus aureus concentration, the expression of p-NF- 魏 B p65 protein increased gradually. The expression of 伪 -SMA and collagen-I protein in Staphylococcus aureus cells treated with different concentrations of Staphylococcus aureus increased significantly, especially in the 105 CFU / mL stimulated group. The expression of 伪 -SMA and collagen-I protein was the highest in the 105 CFU / mL group. PDTC could significantly inhibit the expression of TNF- 伪, IL-1 尾, IL-6 and TGF- 尾 1 mRNA. PDTC could significantly inhibit the expression of 伪 -SMA-collagen-I and p-NF- 魏 B p65 protein. The results suggest that NF- 魏 B signaling pathway plays an important role in the process of transforming the breast fibroblasts into myofibroblasts induced by Staphylococcus aureus. This study lays a foundation for revealing the mechanism of staphylococcus aureus mastitis sclerosis.
【作者单位】：黑龙江八一农垦大学动物科技学院;
【基金】：黑龙江八一农垦大学博士后启动基金资助项目;黑龙江八一农垦大学博士启动基金资助项目(XYB2013-11)
【分类号】：S858.23

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