实用医学杂志
實用醫學雜誌
실용의학잡지
The Journal of Practical Medicine
2015年
17期
2799-2802
,共4页
谢大泽%黄利兴%刘东升%朱俊%谢勇%周南进
謝大澤%黃利興%劉東升%硃俊%謝勇%週南進
사대택%황리흥%류동승%주준%사용%주남진
LX A4%BML-111%RAW264.7细胞%TLR4%NF-κB
LX A4%BML-111%RAW264.7細胞%TLR4%NF-κB
LX A4%BML-111%RAW264.7세포%TLR4%NF-κB
LX A4%BML-111%RAW264.7 macrophage%TLR4%NF-κB
目的：肠道巨噬细胞定位于肠黏膜相关淋巴组织及黏膜固有层，在保持肠道稳态及免疫防御中发挥重要作用。研究脂氧素（LXs）A4及其受体激动剂BML-111对脂多糖（LPS）作用巨噬细胞RAW264．7存活和TLR4／NF-κB信号通路的影响。方法：CCK-8法观察LPS对RAW264．7的毒性作用，RT-qPCR法检测细胞内TLR4、TRAF6 mRNA的表达，Western blot法检测细胞内TLR4、TRAF6和pNF-κB p65的蛋白水平。结果：在1000 ng／mL浓度LPS组，作用6 h后，LX A4组和BML-111组对RAW264．7细胞存活率显著增高（P＜0．05）。在LPS作用下，LX A4组和BML-111组对RAW264．7细胞的TLR4 mRNA及蛋白水平显著高于相对应的无LPS组（P ＜0．05），TRAF6 mRNA的表达均高于相对应的无LPS组（P ＜0．05）；而LX A4组和BML-111组的TRAF6蛋白水平则低于对照组（P＜0．05），高于相对应的无LPS组（P＜0．05）。在LPS作用下，LX A4组和BML-111组pNF-κB p65蛋白水平低于对照组（P ＜0．05），而对照组又高于相对应的无LPS组（P ＜0．05）；且LX A4和BML-111作用无差异（P＞0．05）。结论：LX A4和BML-111能够抑制LPS对RAW264．7细胞的作用及TLR4／NF-κB信号通路的激活，有助减轻炎症反应；性质稳定的BML-111更有望成为IBD治疗的新契机。
目的：腸道巨噬細胞定位于腸黏膜相關淋巴組織及黏膜固有層，在保持腸道穩態及免疫防禦中髮揮重要作用。研究脂氧素（LXs）A4及其受體激動劑BML-111對脂多糖（LPS）作用巨噬細胞RAW264．7存活和TLR4／NF-κB信號通路的影響。方法：CCK-8法觀察LPS對RAW264．7的毒性作用，RT-qPCR法檢測細胞內TLR4、TRAF6 mRNA的錶達，Western blot法檢測細胞內TLR4、TRAF6和pNF-κB p65的蛋白水平。結果：在1000 ng／mL濃度LPS組，作用6 h後，LX A4組和BML-111組對RAW264．7細胞存活率顯著增高（P＜0．05）。在LPS作用下，LX A4組和BML-111組對RAW264．7細胞的TLR4 mRNA及蛋白水平顯著高于相對應的無LPS組（P ＜0．05），TRAF6 mRNA的錶達均高于相對應的無LPS組（P ＜0．05）；而LX A4組和BML-111組的TRAF6蛋白水平則低于對照組（P＜0．05），高于相對應的無LPS組（P＜0．05）。在LPS作用下，LX A4組和BML-111組pNF-κB p65蛋白水平低于對照組（P ＜0．05），而對照組又高于相對應的無LPS組（P ＜0．05）；且LX A4和BML-111作用無差異（P＞0．05）。結論：LX A4和BML-111能夠抑製LPS對RAW264．7細胞的作用及TLR4／NF-κB信號通路的激活，有助減輕炎癥反應；性質穩定的BML-111更有望成為IBD治療的新契機。
목적：장도거서세포정위우장점막상관림파조직급점막고유층，재보지장도은태급면역방어중발휘중요작용。연구지양소（LXs）A4급기수체격동제BML-111대지다당（LPS）작용거서세포RAW264．7존활화TLR4／NF-κB신호통로적영향。방법：CCK-8법관찰LPS대RAW264．7적독성작용，RT-qPCR법검측세포내TLR4、TRAF6 mRNA적표체，Western blot법검측세포내TLR4、TRAF6화pNF-κB p65적단백수평。결과：재1000 ng／mL농도LPS조，작용6 h후，LX A4조화BML-111조대RAW264．7세포존활솔현저증고（P＜0．05）。재LPS작용하，LX A4조화BML-111조대RAW264．7세포적TLR4 mRNA급단백수평현저고우상대응적무LPS조（P ＜0．05），TRAF6 mRNA적표체균고우상대응적무LPS조（P ＜0．05）；이LX A4조화BML-111조적TRAF6단백수평칙저우대조조（P＜0．05），고우상대응적무LPS조（P＜0．05）。재LPS작용하，LX A4조화BML-111조pNF-κB p65단백수평저우대조조（P ＜0．05），이대조조우고우상대응적무LPS조（P ＜0．05）；차LX A4화BML-111작용무차이（P＞0．05）。결론：LX A4화BML-111능구억제LPS대RAW264．7세포적작용급TLR4／NF-κB신호통로적격활，유조감경염증반응；성질은정적BML-111경유망성위IBD치료적신계궤。
Objective To determine the effect of lipoxins (LX) A4 and its agonist (BML-111) on the survival of RAW264.7 macrophage cells and TLR4/NF-κB signaling pathway. Methods RAW264.7 cells were treated with different concentrations of LPS, then the effect of LX A4 and BML-111 on the survival rate of these cells was observed. Cytotoxicity were detected by CCK-8 method and RT-PCR was used to detect the TLR4 and TRAF6 mRNA. The protein levels of TLR4 and pNF-κB p65 in RAW264.7 were determined by Western Blot. Results The survival rates of macrophage treated with LPS for 6 h in 1 000 ng/mL LPS in LX A4 group and BML-111 group were significantly higher than that in control group (P < 0.05). In the present of LPS, the TLR4 mRNA levels in RAW264.7 cells from LX A4 group and BML-111 group were significantly higher than those in the corresponding non-LPS groups. And the TRAF6 mRNA levels in each LPS stimulation group were higher than those in the corresponding non-LPS groups (P<0.05), while the protein level of TRAF6 in LX A4 and BML-111 groups were significantly lower than that in control group (P < 0.05). Stimulated with LPS, the protein levels of pNF-κB p65 in the LX A4 group and BML-111 group were all significantly lower than that in control group (P<0.05), and pNF-κB p65 expression level in control group was also significantly higher than the corresponding non-LPS groups (P < 0.05). Meanwhile, no significant difference was found between LX A4 and BML-111 group (P > 0.05). Conclusion LX A4 and BML-111 could inhibit the cytotoxicity of LPS on the RAW264.7 macrophage cells through the inhibition of the activation of TLR4/NF-κB signaling pathway, then reduce the inflammation. And this stable BML-111 may appear as another promising treatment for IBD disease.
