IFITM3在脓毒症模型及胆碱能抗炎模型中的表达
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摘要
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.
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.
引文
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[20]Li He-ping,Chen Pei-ge,Liu Fu-tao,et al.Characterization and antiinflammation role of swine IFITM3 gene[J].Oncotarget,2017,8(43):73579-73589
[21]Chen Xu-xin,Lu Tang,Feng Jian,et al.Downregulation of Paralemmin-3 Ameliorates Lipopolysaccharide-Induced Acute Lung Injury in Rats by Regulating Inflammatory Response and Inhibiting Formation of TLR4/My D88 and TLR4/TRIF Complexes[J].Oncotarget,2017,8(43):73579-73589
[22]Jiang Li-qun,Xia Tian,Hu Yun-Hong,et al.IFITM3 inhibits virus-triggered induction of type I interferon by mediating autophagosome-dependent degradation of IRF3[J].Cell Mol Immunol,2017Apr 24
[2]Ranjbar S,Haridas V,Jasenosky LD,et al.A Role for IFITM Proteins in Restriction of Mycobacterium tuberculosis Infection[J].Cell Rep,2015,13(5):874-883
[3]Saetre P,Emilsson L,Axelsson E,et al.Inflammation-related genes up-regulated in schizophrenia brains[J].BMC Psychiatry,2007,7:46
[4]Arion D,Unger T,Lewis DA,et al.Molecular Evidence for Increased Expression of Genes Related to Immune and Chaperone Function in the Prefrontal Cortex in Schizophrenia[J].Biol Psychiatry,2007,62(7):711-721
[5]Foster DJ,Jones CK,Conn PJ,et al.Emerging approaches for treatment of schizophrenia:modulation of cholinergic signaling[J].Discov Med,2012,14(79):413-420
[6]Fujii T,Mashimo M,Moriwaki Y,et al.Expression and Function of the Cholinergic System in Immune Cells[J].Front Immunol,2017,8:1085
[7]Wu Xiao-jing,Yang Xu-ming,Song Xue-min,et al.The Role of Erbin in GTS-21 Regulating Inflammatory Responses in MDP-Stimulated Macrophages[J].Shock,2017,47(5):653-657
[8]Conway-Morris A,Wilson J,Shankar-Hari M.Immune Activation in Sepsis[J].Crit Care Clin,2018,34(1):29-42
[9]Avlas O,Fallach R,Shainberg A,et al.Toll-like receptor 4 stimulation initiates an inflammatory response that decreases cardiomyocyte contractility[J].Antioxid Redox Signal,2011,15(7):1895-909
[10]Gil M,Kim YK,Hong SB,et al.Naringin Decreases TNF-αand HMGB1 Release from LPS-Stimulated Macrophages and Improves Survival in a CLP-Induced Sepsis Mice[J].PLo S One,2016,11(10):e0164186
[11]Tracey KJ.The Inflammatory Reflex[J].Nature,2002,420(6917):853
[12]Zoheir N,Lappin DF,Nile CJ,et al.Acetylcholine and the alpha 7nicotinic receptor:a potential therapeutic target for the treatment of periodontal disease[J].Inflamm Res,2012,61(9):915-926
[13]Kox M,Pompe JC,Peters E,et al.α7 nicotinic acetylcholine receptor agonist GTS-21 attenuates ventilator-induced tumour necrosis factor-αproduction and lung injury[J].Br J Anaesth,2011,107(4):559-566
[14]Chatterjee PK,Yeboah MM,Solanki MH,et al.Activation of the cholinergic anti-inflammatory pathway by GTS-21 attenuates cisplatin-induced acute kidney injury in mice[J].PLo S One,2017,12(11):e0188797
[15]Kong Wei-lan,Kang Kai,Gao Yang,et al.Dexmedetomidine alleviates LPS-induced septic cardiomyopathy via the cholinergic anti-inflammatory pathway in mice[J].Am J Transl Res,2017,9(11):5040-5047
[16]Egea J,Buendia I,Parada E,et al.Anti-inflammatory role of microglial alpha7 n ACh Rs and its role in neuroprotection[J].Biochem Pharmacol,2015,97(4):463-472
[17]Zhu Ya-juan,Peng Ke,Meng Xiao-wen,et al.Attenuation of neuroinflammation by dexmedetomidine is associated with activation of a cholinergic anti-inflammatory pathway in a rat tibial fracture model[J].Brain Res,2016,1644:1-8
[18]Song Qi,Hu Sen,Wang Hai-bin,et al.Electroacupuncturing at Zusanli point(ST36)attenuates pro-inflammatory cytokine release and organ dysfunction by activating cholinergic anti-inflammatory pathway in rat with endotoxin challenge[J].Afr J Tradit Complement Altern Med,2014,11(2):469-474
[19]Gao Yang,Kai Kang,Liu Hai-tao,et al.GTS-21 attenuates LPS-induced renal injury via the cholinergic anti-inflammatory pathway in mice[J].Am J Transl Res,2017,9(10):4673-4681
[20]Li He-ping,Chen Pei-ge,Liu Fu-tao,et al.Characterization and antiinflammation role of swine IFITM3 gene[J].Oncotarget,2017,8(43):73579-73589
[21]Chen Xu-xin,Lu Tang,Feng Jian,et al.Downregulation of Paralemmin-3 Ameliorates Lipopolysaccharide-Induced Acute Lung Injury in Rats by Regulating Inflammatory Response and Inhibiting Formation of TLR4/My D88 and TLR4/TRIF Complexes[J].Oncotarget,2017,8(43):73579-73589
[22]Jiang Li-qun,Xia Tian,Hu Yun-Hong,et al.IFITM3 inhibits virus-triggered induction of type I interferon by mediating autophagosome-dependent degradation of IRF3[J].Cell Mol Immunol,2017Apr 24