胸腺肽β4通过丝裂原活化蛋白激酶信号通路致敏肥大细胞的研究
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摘要
目的研究胸腺肽β4(Tβ4)对肥大细胞的致敏作用及可能的机制。方法将不同浓度的Tβ4作用于肥大细胞株RBL-2H3,检测肥大细胞释放的β氨基己糖苷酶。用丝裂原活化蛋白激酶(MAPK)信号通路特异性抑制剂10μmol·L~(-1)U0126或抗过敏剂5 mmol·L~(-1)色甘酸钠分别联合4 pmol·L~(-1)Tβ4作用于小鼠肠黏膜肥大细胞,检测是否可以阻断Tβ4对肥大细胞β氨基己糖苷酶的释放作用。结果随着Tβ4浓度的增加对肥大细胞的脱颗粒效应逐渐增加,在4 pmol·L~(-1)时达到峰值。同样,4 pmol·L~(-1)的Tβ4可显著引起肠道肥大细胞的过敏反应,U0126和色甘酸钠可以抑制Tβ4的致敏效应。结论 Tβ4可通过MAPK信号通路致敏肠黏膜肥大细胞,但可被U0126和色甘酸钠阻断。这为Tβ4的药物开发和应用提供参考依据。
Objective To investigate the effect of thymosin β4( Tβ4)activating mast cells and its possible mechanism. Methods To observe the activation of mast cells,mast cells RBL-2 H3 and mouse intestinal mucosae were stimulated by Tβ4 at various concentrations,then the release of β-hexosaminidase was determined. In addition,mitogen-activated protein kinase( MAPK) specific inhibitor U0126( 10 μmol·L~(-1))or disodium cromoglycate( 5 mmol·L~(-1)) was used to block the activation of mice intestinal mucosal mast cells induced by Tβ4( 4 pmol·L~(-1)). Results In this study,we found that the degranulation effect of mast cells was gradually increased with the dose of Tβ4,peaking at 4 pmol·L~(-1). Similarly,Tβ4( 4 pmol·L~(-1)) caused the sensitivity of intestinal mast cells. However,the effect of Tβ4 on intestinal mast cells could be inhibited by U0126 and disodium cromoglycate. Conclusion Tβ4 sensitizes the intestinal mucosal mast cells through MAPK signaling pathway,which can be blocked by disodium cromoglycate. This will provide a reference for the drug development and administration of Tβ4.
Objective To investigate the effect of thymosin β4( Tβ4)activating mast cells and its possible mechanism. Methods To observe the activation of mast cells,mast cells RBL-2 H3 and mouse intestinal mucosae were stimulated by Tβ4 at various concentrations,then the release of β-hexosaminidase was determined. In addition,mitogen-activated protein kinase( MAPK) specific inhibitor U0126( 10 μmol·L~(-1))or disodium cromoglycate( 5 mmol·L~(-1)) was used to block the activation of mice intestinal mucosal mast cells induced by Tβ4( 4 pmol·L~(-1)). Results In this study,we found that the degranulation effect of mast cells was gradually increased with the dose of Tβ4,peaking at 4 pmol·L~(-1). Similarly,Tβ4( 4 pmol·L~(-1)) caused the sensitivity of intestinal mast cells. However,the effect of Tβ4 on intestinal mast cells could be inhibited by U0126 and disodium cromoglycate. Conclusion Tβ4 sensitizes the intestinal mucosal mast cells through MAPK signaling pathway,which can be blocked by disodium cromoglycate. This will provide a reference for the drug development and administration of Tβ4.
引文
[1] GOLDSTEIN A L,KLEINMAN H K. Advances in the basic and clinical applications of thymosin beta 4[J]. Expert Opin Biol Ther,2015,15(Suppl 1):S139-S145.
[2] MICERA A,BONINI S,LAMBIASE A,et al. Conjunctival expression of thymosin-beta 4 in vernal keratoconjunctivitis[J]. Mol Vis,2006,12:1594-1600.
[3] WEI M,DUAN D,LIU Y,et al. Increased thymosin beta 4 levels in the serum and SF of knee osteoarthritis patients correlate with disease severity[J]. Regul Pept,2013,185(1):34-36.
[4] BAHRI R,CUSTOVIC A,KOROSEC P,et al. Mast cell activation test in the diagnosis of allergic disease and anaphylaxis[J]. J Alleregy Clin Immun,2018,142(2):485-496. e16.
[5] DONG H,ZHANG X,WANG Y,et al. Suppression of brain mast cells degranulation inhibits microglial activation and central nervous system inflammation[J]. Mol Neurobiol,2017,54(2):997-1007.
[6] CROCKFORD D,TURJMAN N,ALLAN C,et al. Thymosin beta4:structure,function,and biological properties supporting current and future clinical applications[J]. Ann N Y Acad Sci,2010,1194(1):179-189.
[7] DUNN S P,HEIDEMANN D G,CHOW C Y,et al. Treatment of chronic nonhealing neurotrophic corneal epithelial defects with thymosin beta 4[J]. Ann N Y Acad Sci,2010,1194(1):199-206.
[8] SOSNE G,OUSLER G W. Thymosin beta 4 ophthalmic solution for dry eye:a randomized,placebo-controlled,PhaseⅡclinical trial conducted using the controlled adverse environment(CAETM)model[J/OL]. Clin Ophthalmol,2015,9:877-884. 2015-05-20[2018-05-17]. http://europepmc. org/articles/PMC4445951.
[9] FASOLATO C,HOTH M,MATTHEWS G,et al. Ca2+and Mn2+influx through receptor-mediated activation of nonspecific cation channels in mast cells[J]. Proc Natl Acad Sci U S,1993,90(7):3068-3072.
[10] SAMAYAWARDHENA L A,KAPUR R,CRAIG A W. Involvement of Fyn kinase in Kit and integrin-mediated Rac activation,cytoskeletal reorganization,and chemotaxis of mast cells[J].Blood,2007,109(9):3679-3686.
[11] DUAN W,CHAN J H,WONG C H,et al. Anti-inflammatory effects of mitogen-activated protein kinase kinase inhibitor U0126in an asthma mouse model[J]. J Immunol,2004,172(11):7053-7059.
[12] SPATARO A C,BOSMANN H B. Mechanism of action of disodium cromoglycate-mast cell calcium ion influx after a histamine-releasing stimulus[J]. Biochem Pharmacol,1976,25(5):505-510.
[13] ARIDOR M,RAJMILEVICH G,BEAVEN M A,et al. Activation of exocytosis by the heterotrimeric G protein Gi3[J]. Science,1993,262(5139):1569-1572.
[2] MICERA A,BONINI S,LAMBIASE A,et al. Conjunctival expression of thymosin-beta 4 in vernal keratoconjunctivitis[J]. Mol Vis,2006,12:1594-1600.
[3] WEI M,DUAN D,LIU Y,et al. Increased thymosin beta 4 levels in the serum and SF of knee osteoarthritis patients correlate with disease severity[J]. Regul Pept,2013,185(1):34-36.
[4] BAHRI R,CUSTOVIC A,KOROSEC P,et al. Mast cell activation test in the diagnosis of allergic disease and anaphylaxis[J]. J Alleregy Clin Immun,2018,142(2):485-496. e16.
[5] DONG H,ZHANG X,WANG Y,et al. Suppression of brain mast cells degranulation inhibits microglial activation and central nervous system inflammation[J]. Mol Neurobiol,2017,54(2):997-1007.
[6] CROCKFORD D,TURJMAN N,ALLAN C,et al. Thymosin beta4:structure,function,and biological properties supporting current and future clinical applications[J]. Ann N Y Acad Sci,2010,1194(1):179-189.
[7] DUNN S P,HEIDEMANN D G,CHOW C Y,et al. Treatment of chronic nonhealing neurotrophic corneal epithelial defects with thymosin beta 4[J]. Ann N Y Acad Sci,2010,1194(1):199-206.
[8] SOSNE G,OUSLER G W. Thymosin beta 4 ophthalmic solution for dry eye:a randomized,placebo-controlled,PhaseⅡclinical trial conducted using the controlled adverse environment(CAETM)model[J/OL]. Clin Ophthalmol,2015,9:877-884. 2015-05-20[2018-05-17]. http://europepmc. org/articles/PMC4445951.
[9] FASOLATO C,HOTH M,MATTHEWS G,et al. Ca2+and Mn2+influx through receptor-mediated activation of nonspecific cation channels in mast cells[J]. Proc Natl Acad Sci U S,1993,90(7):3068-3072.
[10] SAMAYAWARDHENA L A,KAPUR R,CRAIG A W. Involvement of Fyn kinase in Kit and integrin-mediated Rac activation,cytoskeletal reorganization,and chemotaxis of mast cells[J].Blood,2007,109(9):3679-3686.
[11] DUAN W,CHAN J H,WONG C H,et al. Anti-inflammatory effects of mitogen-activated protein kinase kinase inhibitor U0126in an asthma mouse model[J]. J Immunol,2004,172(11):7053-7059.
[12] SPATARO A C,BOSMANN H B. Mechanism of action of disodium cromoglycate-mast cell calcium ion influx after a histamine-releasing stimulus[J]. Biochem Pharmacol,1976,25(5):505-510.
[13] ARIDOR M,RAJMILEVICH G,BEAVEN M A,et al. Activation of exocytosis by the heterotrimeric G protein Gi3[J]. Science,1993,262(5139):1569-1572.