FTY720抑制TLR4信号转导的炎症反应
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摘要
FTY720是一种1-磷酸神经鞘氨醇(sphingosine 1-phosphate,S1P)受体拮抗剂。为阐明FTY720抑制炎症反应的机理,本研究采用流式细胞术检测FTY720对细菌脂多糖(LPS)刺激后小鼠脾脏细胞TLR4表达水平的影响。研究发现,注射适当剂量的FTY720可削弱LPS所引起的TLR4表达水平增强(FTY720处理组与对照组相比,TLR4+细胞构成比分别为6.7%±1.5%和38.6%±3.1%,P<0.01),并可消除LPS刺激所引起的外周血TNF-α、IFN-γ和IL-12等Th1型细胞因子表达水平增高(FTY720组血清TNF-α、IFN-γ和IL-12浓度分别为(72±22)ng/ml、(46±21)ng/ml和(19±4)ng/ml,LPS组分别为(300±75)ng/ml、(175±35)ng/ml和(50±8)ng/ml,均为P<0.01)。相反,FTY720组血清Th2型细胞因子IL-4水平与对照组相比分别为(7±3)ng/ml和(7±2)ng/ml,无显著性差异。在体外细胞培养实验中可观察到一致的结果。这些发现提示,FTY720可能通过抑制LPS/TLR4信息转导通路,下调TNF-α、IFN-γ和IL-12等Th1型细胞因子表达而抑制炎症反应。
FTY720(2-amino-2-[2-(4-octylphenyl)ethyl]propane-1,3-diol) is known to be an agonist of sphingosine 1-phosphate(S1P) receptor.To make clear the mechanism by which FTY720 inhibits inflamation,flow cytometry was performed to determine the effect of FTY720 on LPS-induced up-regulation of TLR4 in murine spleen cells in this study.It showed that adequate dosage of FTY720 abrogated LPS-induced TLR4 expression.The proportion of TLR4+ cells was 6.7%±1.5% in FTY720-treated group,significantly lower than that in the control group(38.6%±3.1%;P<0.01).Subsequently,up-regulated expression of Th1-type cytokines,including TNF-α,IFN-γ and IL-12,in response to LPS stimulation was abrogated by FTY720 administration.Serum level of TNF-α,IFN-γ and IL-12 were(72±22)ng/ml,(46±21)ng/ml,and(19±4)ng/ml respectively in FTY720 group,significantly lower than that in the control group [(300±75)ng/ml,(175±35)ng/ml,and(50±8) ng/ml,respectively;P<0.01 for all].In contrast,no significant difference was observed in the serum level of Th2 type cytokine IL-4(7 3 ng/ml in the FTY720-treated group and 7 2 ng/ml in LPS-treated group).A similar trend was observed in in vitro cell culture systems.These results suggest that FTY720 may inhibit LPS/TLR4 pathway,down-regulate Th1 type cytokine expression,and modulate inflammatory response.
FTY720(2-amino-2-[2-(4-octylphenyl)ethyl]propane-1,3-diol) is known to be an agonist of sphingosine 1-phosphate(S1P) receptor.To make clear the mechanism by which FTY720 inhibits inflamation,flow cytometry was performed to determine the effect of FTY720 on LPS-induced up-regulation of TLR4 in murine spleen cells in this study.It showed that adequate dosage of FTY720 abrogated LPS-induced TLR4 expression.The proportion of TLR4+ cells was 6.7%±1.5% in FTY720-treated group,significantly lower than that in the control group(38.6%±3.1%;P<0.01).Subsequently,up-regulated expression of Th1-type cytokines,including TNF-α,IFN-γ and IL-12,in response to LPS stimulation was abrogated by FTY720 administration.Serum level of TNF-α,IFN-γ and IL-12 were(72±22)ng/ml,(46±21)ng/ml,and(19±4)ng/ml respectively in FTY720 group,significantly lower than that in the control group [(300±75)ng/ml,(175±35)ng/ml,and(50±8) ng/ml,respectively;P<0.01 for all].In contrast,no significant difference was observed in the serum level of Th2 type cytokine IL-4(7 3 ng/ml in the FTY720-treated group and 7 2 ng/ml in LPS-treated group).A similar trend was observed in in vitro cell culture systems.These results suggest that FTY720 may inhibit LPS/TLR4 pathway,down-regulate Th1 type cytokine expression,and modulate inflammatory response.
引文
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[12]Tsunemi S,Iwasaki T,Kitano S,et al.Effects of the novelimmunosuppressant FTY720in a murine rheumatoid arthritismodel[J].Clin Immunol,2010,136:197-204.
[13]Imado T,Iwasaki T,Kitano S,et al.The protective role ofhost Toll-like receptor-4in acute graft-versus-host disease[J].Transplantation,2010,90:1063-1070.
[14]Nicoletti F,Auci DL,Manqano K,et al.5-androstenediolameliorates pleurisy,septic shock,and experimental autoim-mune encephalomyelitis in mice[J].Autoimmu Dis,2010,2010:757432.
[15]Singh M,Cugati G,Singh P,et al.Fingolimod:The first o-ral drug approved by food and drug administration;A break-through in treatment of multiple sclerosis[J].J PharmBioallied Sci,2011 3:460-461.
[16]Romero Rosales K,Singh G,Wu K,et al.Sphingolipid-baseddrugs selectively kill cancer cells by down-regulating nutrienttransporter proteins[J].Biochem J,2011,439:299-311.
[2]Akira S,Takeda K.Toll-like receptor signaling[J].Nat RevImmunol,2004,4:499-511.
[3]Matloubian M,Lo CG,Cinamon G,et al.Lymphocyteegress from thymus and peripheral lymphoid organs is de-pendent on S1Preceptor 1[J].Nature,2004,427:355-360.
[4]Zhang Z,Schluesener HJ.FTY720:a most promising immu-nosuppressant modulating immune cell functions[J].MiniRev Med Chem,2007,7:845-850.
[5]Brinkmann V.Sphingosine 1-phosphate receptors in healthand disease:mechanistic insights from gene deletion studiesand reverse 5pharmacology[J].Pharmacol Ther,2007,115:84-105.
[6]Lin Y,Xie M,Chen Y,et al.Preterm delivery induced byLPS in syngeneically impregnated BALB/c and NOD/SCIDmice[J].J Reprod Immunol,2006,71:87-101.
[7]Sehrawat S,Rouse BT.Anti-inflammatory effects of FTY720against viral-induced immunopathology:role of drug-inducedconversion of T cells to become Foxp3+regulators[J].J Im-munol,2008,180:7636-7647.
[8]Lin Y,Ren L,Wang W,et al.Effect of TLR3and TLR7ac-tivation in uterine NK cells from non-obese diabetic(NOD)mice[J].J Reprod Immunol,2009,82:12-23.
[9]仲艳敏,林羿,李莉平,等.人工合成相关寡肽抑制poly I:C诱发性流产的作用[J].现代免疫学,2007,27:145-150.
[10]Lin Y,Zhong Y,Shen W,et al.TSLP-induced placental DCactivation and IL-10+NK cell expansion:Comparative studybased on BALB/c×C57BL/6and NOD/SCID×C57BL/6pregnant models[J].Clin Immunol,2008,126:104-117.
[11]Yoshida Y,Tsuji T,Fujita T,et al.Relapse of experimentalautoimmune encephalomyelitis after discontinuation ofFTY720(Fingolimod)treatment,but not after combinationof FTY720and pathogenic autoantigen[J].Biol Pharm Bull,2011,34:933-936.
[12]Tsunemi S,Iwasaki T,Kitano S,et al.Effects of the novelimmunosuppressant FTY720in a murine rheumatoid arthritismodel[J].Clin Immunol,2010,136:197-204.
[13]Imado T,Iwasaki T,Kitano S,et al.The protective role ofhost Toll-like receptor-4in acute graft-versus-host disease[J].Transplantation,2010,90:1063-1070.
[14]Nicoletti F,Auci DL,Manqano K,et al.5-androstenediolameliorates pleurisy,septic shock,and experimental autoim-mune encephalomyelitis in mice[J].Autoimmu Dis,2010,2010:757432.
[15]Singh M,Cugati G,Singh P,et al.Fingolimod:The first o-ral drug approved by food and drug administration;A break-through in treatment of multiple sclerosis[J].J PharmBioallied Sci,2011 3:460-461.
[16]Romero Rosales K,Singh G,Wu K,et al.Sphingolipid-baseddrugs selectively kill cancer cells by down-regulating nutrienttransporter proteins[J].Biochem J,2011,439:299-311.