%0 Journal Article %T <i>LBP</i>基因沉默对LPS诱导水牛单核巨噬细胞炎症相关基因表达的影响<br>Effects of LBP Gene Silencing on Gene Expression in Water Buffalo Monocyte-Macrophages Post-LPS Stimulation %A 谢亮亮 %A 黄时海 %A 汤龙 %A 韦英明 %A 石德顺 %A 李湘萍< %A br> %A XIE Liang-liang %A HUANG Shi-hai %A TANG Long %A WEI Ying-ming %A SHI De-shun %A LI Xiang-ping %J 西南大学学报(自然科学版) %D 2018 %R 10.13718/j.cnki.xdzk.2018.07.002 %X 为了探讨抑制脂多糖结合蛋白（LBP）基因表达对水牛单核巨噬细胞在内毒素（LPS）诱导下炎症相关基因的表达，首先采用三质粒慢病毒包装系统包装<i>LBP</i>基因<i>shRNA</i>重组质粒pSicoR-GFP-shLBP774.利用病毒颗粒感染水牛单核巨噬细胞，进行荧光定量qRT-PCR及ELISA检测.结果显示，单核巨噬细胞在用5×10<sup>8</sup> IU/mL滴度的LBP-shRNA774慢病毒颗粒，感染指数（MOI）为300、感染7 d的条件下，感染效率超过50%.qRT-PCR检测结果显示，与LPS对照组相比，shLBP774感染组可极显著抑制<i>LBP</i>基因的表达（<i>p</i> < 0.01），<i>CD</i>14，<i>TLR</i>4，<i>TNF</i>-<i>α</i>，<i>IL</i>-1<i>β</i>，<i>IL</i>-8等基因表达显著降低（<i>p</i> < 0.05）.ELISA检测结果发现，与对照组相比，细胞分泌的TNF-α，IL-1β蛋白量显著降低（<i>p</i> < 0.05）.以上结果表明，抑制<i>LBP</i>基因表达可以有效降低LPS诱导下单核巨噬细胞炎症相关基因的表达，进而调控LPS引发的炎症反应，提示可将<i>LBP</i>作为靶基因深入研究水牛单核巨噬细胞免疫功能和LPS致炎信号传导分子机制.<br>In order to investigate the effect of inhibition of LBP (lipopolysaccharide binding protein) gene expression on LPS-induced expression of macrophage inflammatory genes in buffalo mononuclear macrophages, LBP-shRNA 774 recombinant plasmid was used to package the virus, and the suitable conditions for target cell infection were explored. Theninflammation-related genes in virus-infected cells were detected with the qRT-PCR and ELISA methods. The results showed that the virus titer of LBP-shRNA774 was > 5×10<sup>8</sup> IU/mL, MOI was 300, and after 7 days, the infection rate was more than 50%. qRT-PCR showed that compared with the LPS control group, the mRNA level of LBP gene was reduced by 50%, meanwhile CD14, TLR4, TNF-α, IL-1β and IL-8 were decreased significantly (<i>p</i> < 0.05). ELISA demonstrated that the contents of cytokine TNF-α and of IL-1β in the LBP-shRNA774 group were significantly lower than in the LPS control group (<i>p</i> < 0.05). Also, the content of soluble LBP (s LBP) protein was significantly lower (<i>p</i> < 0.05), only accounting for 48.54%. The above results suggested that inhibition of LBP gene expression can effectively reduce the expression of LPS-induced inflammation-related genes of monocyte-macrophage, and regulate the LPS-induced inflammatory response %K 脂多糖结合蛋白 %K RNAi %K 水牛 %K 单核巨噬细胞 %K LPS %K 基因表达< %K br> %K lipopolysaccharide binding protein (LBP) %K RNAi %K water buffalo (< %K i> %K Bubalus bubalis< %K /i> %K ) %K monocyte-macrophages %K lipopolysaccharides (LPS) %K gene expression %U http://xbgjxt.swu.edu.cn/jsuns/html/jsuns/2018/7/20180702.htm