IFITM3在脓毒症模型及胆碱能抗炎模型中的表达

Expression of IFITM3 in Sepsis and Cholinergic Anti-inflammatory Pathway

目的:探讨干扰素诱导的跨膜转运蛋白3(IFITM3)在LPS刺激的RAW264.7细胞系的脓毒症模型中的表达以及胆碱能抗炎模型中的表达。方法:用1μg/mL LPS刺激RAW264.7细胞24、48、72 h后,用Western-Blot法检测各组细胞IFITM3蛋白表达水平。用1μg/mL LPS刺激RAW264.7细胞后,给予50μM胆碱能受体激动剂GTS-21以及同时给予100 n M胆碱能受体拮抗剂α-BGT刺激细胞24 h后,用Western-Blot法检测各组细胞IFITM3蛋白表达水平。用ELISA法检测IL-1β的方法验证脓毒症模型和胆碱能抗炎模型的建立。结果:(1)1μg/mL LPS刺激RAW264.7细胞后,IFITM3蛋白表达明显降低(P<0.01)。(2)1μg/mL LPS刺激RAW264.7细胞后再给予50μM GTS-21,IFITM3蛋白表达明显升高(P<0.001);而给予100 nMα-BGT后,IFITM3蛋白表达明显降低(P<0.001)。结论:LPS刺激的RAW264.7细胞IFITM3蛋白表达降低。给予胆碱能激动剂GTS-21后能够逆转LPS诱导的IFITM3表达的降低,给予胆碱能受体拮抗剂α-BGT则能阻断这种现象。IFITM3有可能在脓毒症中发挥保护作用,并且参与了胆碱能抗炎通路抗炎过程的调节。

Objective： To investigate the expression of IFITM3 in RAW264.7 cell line stimulated by LPS and Cholinergic anti-inflammatory pathway model. Methods： RAW264.7 cell was treated by 1 μg/mL LPS for 24, 48 and 72 h, the protein expression of IFITM3 was determined by Western Blot. 50 μM Cholinergic receptor agonist GTS-21 combined with 100 n M Cholinergic receptor antagonist α-BGT were given after 1 μg/mL LPS stimulation and then the protein expression levels of IFITM3 were determined. The model of sepsis and Cholinergic anti-inflammatory pathway were verified by IL-1β level detection. Results：（1）The expression of IFITM3 in RAW264.7 cell lines were greatly decreased after 1 μg/mL LPS stimulation（P〈0.01）.（2）The expression of IFITM3 was significantly increased after treatment with 50 μM GTS-21 for 24 h（P〈0.001） but notably declined after treatment of 100 nM α-BGT for 24 h following1 μg/mL LPS stimulation（P〈0.001）. Conclusions： The protein expression of IFITM3 was decreased in RAW264.7 cell induced by LPS.Cholinergic receptor agonist GTS-21 could reverse the reduction of IFITM3 induced by LPS, and this effect was abol-ished by Cholinergic receptor antagonist α-BGT. IFITM3 might have protective effect in sepsis and regulate anti-inflammatory process in Cholinergic antiinflammatory pathway.