大麻素受体-2激动剂AM1241对急性肝损伤小鼠肝组织炎性体NLRP3影响
|
摘要
目的探讨大麻素CB2受体激动剂AM1241对刀豆蛋白A(Con A)所致急性小鼠肝损伤影响及机制。方法随机将40只8周龄C57BL/6J雄性小鼠分为对照组、模型组、激动剂(AM1241)3、12 mg/kg组(于造模前1 h分别腹腔注射AM1241 3、12 mg/kg),除对照组外,其余组小鼠尾静脉注射Con A 20 mg/kg复制小鼠急性肝损伤模型,对照组采用相同溶剂和方法。造模8 h后留取血清检测丙氨酸转氨酶(ALT),Western blot法分别检测炎性体3相关核苷酸结合寡聚化结构域样受体(NLRs)、凋亡相关斑点样蛋白(ASC)、半胱氨酸天冬氨酸蛋白酶–1(caspase-1)蛋白的表达量,利用试剂盒检测肝组织匀浆中白细胞介素1β(IL-1β)水平。结果与对照组[(28.5±7.4)U/L]比较,模型组小鼠血清ALT水平[(6 132.5±1 300.8)U/L]明显升高(P <0.05),肝组织中NLRP3、ASC、caspase-1蛋白表达均明显增强(P <0.05);与模型组比较,AMl241 3、12 mg/kg组小鼠血清ALT水平[分别为(2 696.5±956.3)、(534.6±128.6)U/L]明显降低(P <0.05),肝组织中NLRP3、ASC、caspase-1蛋白表达均有所降低(P <0.05);与对照组比较,AMl241组小鼠肝组织中IL-1β含量也明显降低(P <0.05)。结论 CB2受体激动剂AMl241对Con A所致急性小鼠肝损伤有一定的拮抗作用,其机制可能与AM1241抑制小鼠肝组织中NLRP3、ASC、caspase-1蛋白表达,减少肝组织中炎性因子IL-1β含量有关。
Objective To investigate the effect of cannabinoid receptor-2(CB2) agonist AM1241 on nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3) and interleukin-1β(IL-1β) in liver tissue of mice with acute liver injury induced by concanavalin A(Con-A). Methods Forty 8-week-old C57 BL/6 J male mice were randomly divided into a control, a model, and two CB2 receptor agonist(AM1241) groups injected with AM1241 intraperitoneally at doses of 3 and12 mg/kg one hour before the establishment of acute liver injury model with caudal vein injection of 20 mg/kg Con A in all the mice except for the control mice only with intraperitoneal injection of solvent. Eight hours after the injection of Con A,following indicators were detected for all the mice: serum alanine aminotransferase(ALT); inflammatory body 3 related nucleotide oligomerization domain-like receptor protein 3(NLRP3), apoptosis-associated speck-like protein containing a caspase recruitment domain(ASC) and caspase-1 in liver tissues with Western blot; and expression level of interleukin-1 beta(IL-1β) in liver homogenate with enzyme-linked immunosorbent assay(ELISA). Results Compared to those in the control mice, significantly increased serum ALT(6 132.5 ± 1 300.8 vs. 28.5 ± 7.4 U/L, P < 0.05), NLRP3, ASC, and caspase-1 in liver tissues(P < 0.05 for all) were detected in the model mice. In the mice administered with AMl241, significantly decreased serum ALT(2 696.5 ± 956.3 and 534.6 ± 128.6 U/L, both P < 0.05), NLRP3, ASC and caspase-1 protein in liver tissues(all P < 0.05) were detected in comparison with those in the model mice. Significantly decreased IL-1β in liver homogenate was also detected in the mice with AMl241 treatment compared to that in the control mice(P < 0.05).Conclusion CB2 receptor agonist AMl241 may have protective effects on Con A-induced acute liver injury in mice and the effects may be related to inhibitions of NLRP3, ASC, caspase-1, and IL-1β expression in liver tissue of mice.
Objective To investigate the effect of cannabinoid receptor-2(CB2) agonist AM1241 on nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3) and interleukin-1β(IL-1β) in liver tissue of mice with acute liver injury induced by concanavalin A(Con-A). Methods Forty 8-week-old C57 BL/6 J male mice were randomly divided into a control, a model, and two CB2 receptor agonist(AM1241) groups injected with AM1241 intraperitoneally at doses of 3 and12 mg/kg one hour before the establishment of acute liver injury model with caudal vein injection of 20 mg/kg Con A in all the mice except for the control mice only with intraperitoneal injection of solvent. Eight hours after the injection of Con A,following indicators were detected for all the mice: serum alanine aminotransferase(ALT); inflammatory body 3 related nucleotide oligomerization domain-like receptor protein 3(NLRP3), apoptosis-associated speck-like protein containing a caspase recruitment domain(ASC) and caspase-1 in liver tissues with Western blot; and expression level of interleukin-1 beta(IL-1β) in liver homogenate with enzyme-linked immunosorbent assay(ELISA). Results Compared to those in the control mice, significantly increased serum ALT(6 132.5 ± 1 300.8 vs. 28.5 ± 7.4 U/L, P < 0.05), NLRP3, ASC, and caspase-1 in liver tissues(P < 0.05 for all) were detected in the model mice. In the mice administered with AMl241, significantly decreased serum ALT(2 696.5 ± 956.3 and 534.6 ± 128.6 U/L, both P < 0.05), NLRP3, ASC and caspase-1 protein in liver tissues(all P < 0.05) were detected in comparison with those in the model mice. Significantly decreased IL-1β in liver homogenate was also detected in the mice with AMl241 treatment compared to that in the control mice(P < 0.05).Conclusion CB2 receptor agonist AMl241 may have protective effects on Con A-induced acute liver injury in mice and the effects may be related to inhibitions of NLRP3, ASC, caspase-1, and IL-1β expression in liver tissue of mice.
引文
[1]Zhang Y,Hu K.Rethinking the pathogenesis of hepatitis B virus(HBV)infection[J].Journal of Medical Virology,2015,87(12):1989-1999.
[2]Dienes HP,Drebber U.Pathology of immune-mediated liver injury[J].Dig Dis,2010,28(1):57-62.
[3]Kersse K,Bertrand MJ,Lamkanfi M,et al.NOD-like receptors and the innate immune system:coping with danger,damage and death[J].Cytokine and Growth Factor Reviews,2011,22(5-6):257.
[4]Paolo C,Marco D,Ferdinando G,et al.The role of the endocannabinoid system in liver diseases[J].Best Practice and Research Clinical Endocrinology and Metabolism,2009,23:65-77.
[5]Pacher P,Gao B.Endocannabinoids and liver disease.III.Endocannabinoid effects on immune cells:implications for inflammatory liver diseases[J].Am J Physiol Gastrointest Liver Physiol,2008,294(4):G850.
[6]Sándor B,Douglas OH,Pan H.et al Cannabinoid-2 receptor mediates protection against hepatic ischemia/reperfusion injury[J].FASEB,2007,21(8):1788-1800.
[7]何萍,吴雅峰,王豫萍,等.大麻素受体-2激动剂AM1241对肝纤维化小鼠肝组织血小板衍生生长因子表达的影响[J].中华肝脏病杂志,2017,25(11):841-846.
[8]Manns MP,Czaja AJ,Gorham JD,et al.Diagnosis and management of autoimmune hepatitis[J].Hepatology,2010,19(1):2193-2213.
[9]Wang QX,Yan L,Ma X.Autoimmune hepatitis in the Asia-Pacific area[J].Journal of Clinical and Translational Hepatology,2018,6(1):48-56.
[10]Sass G,Heinlein S,Agli A,et al.Cytokine expression in three mouse models of experimental hepatitis[J].Cytokine,2002,19(3):115-120.
[11]Malfitano AM,Basu S,Maresz K,et al.What we know and do not know about the cannabinoid receptor 2(CB2)[J].Seminars in Immunology,2014,26(5):369-379.
[12]Leleuchavain N,Desreumaux P,Chavatte P,et al.Therapeutical potential of CB2 receptors in immune-related diseases[J].Current Molecular Pharmacology,2013,6(3):183.
[13]Georgiana BS,Erawan BK,Lidia T,et al.Expression analysis of inflammasomes in experimental models of inflammatory and fibrotic liver disease[J].Journal of Inflammation(London,England),2012,9(1):49.
[14]张向颖,任锋,段钟平.炎症小体在肝脏疾病致病机制中作用的研究进展[J].中华肝脏病杂志,2014,22(6):478-480.
[15]Zielinski CE,Mele F,Aschenbrenner D,et al.Pathogen-induced human TH17 cells produce IFN-γor IL-10 and are regulated by IL-1β[J].Nature,2012,484(7395):514.
[16]García JA,Volt H,Venegas C,et al.Disruption of the NF-κB/NLRP3 connection by melatonin requires retinoid-related orphan receptor-αand blocks the septic response in mice[J].Faseb Journal,2015,29(9):3863-3875.
[2]Dienes HP,Drebber U.Pathology of immune-mediated liver injury[J].Dig Dis,2010,28(1):57-62.
[3]Kersse K,Bertrand MJ,Lamkanfi M,et al.NOD-like receptors and the innate immune system:coping with danger,damage and death[J].Cytokine and Growth Factor Reviews,2011,22(5-6):257.
[4]Paolo C,Marco D,Ferdinando G,et al.The role of the endocannabinoid system in liver diseases[J].Best Practice and Research Clinical Endocrinology and Metabolism,2009,23:65-77.
[5]Pacher P,Gao B.Endocannabinoids and liver disease.III.Endocannabinoid effects on immune cells:implications for inflammatory liver diseases[J].Am J Physiol Gastrointest Liver Physiol,2008,294(4):G850.
[6]Sándor B,Douglas OH,Pan H.et al Cannabinoid-2 receptor mediates protection against hepatic ischemia/reperfusion injury[J].FASEB,2007,21(8):1788-1800.
[7]何萍,吴雅峰,王豫萍,等.大麻素受体-2激动剂AM1241对肝纤维化小鼠肝组织血小板衍生生长因子表达的影响[J].中华肝脏病杂志,2017,25(11):841-846.
[8]Manns MP,Czaja AJ,Gorham JD,et al.Diagnosis and management of autoimmune hepatitis[J].Hepatology,2010,19(1):2193-2213.
[9]Wang QX,Yan L,Ma X.Autoimmune hepatitis in the Asia-Pacific area[J].Journal of Clinical and Translational Hepatology,2018,6(1):48-56.
[10]Sass G,Heinlein S,Agli A,et al.Cytokine expression in three mouse models of experimental hepatitis[J].Cytokine,2002,19(3):115-120.
[11]Malfitano AM,Basu S,Maresz K,et al.What we know and do not know about the cannabinoid receptor 2(CB2)[J].Seminars in Immunology,2014,26(5):369-379.
[12]Leleuchavain N,Desreumaux P,Chavatte P,et al.Therapeutical potential of CB2 receptors in immune-related diseases[J].Current Molecular Pharmacology,2013,6(3):183.
[13]Georgiana BS,Erawan BK,Lidia T,et al.Expression analysis of inflammasomes in experimental models of inflammatory and fibrotic liver disease[J].Journal of Inflammation(London,England),2012,9(1):49.
[14]张向颖,任锋,段钟平.炎症小体在肝脏疾病致病机制中作用的研究进展[J].中华肝脏病杂志,2014,22(6):478-480.
[15]Zielinski CE,Mele F,Aschenbrenner D,et al.Pathogen-induced human TH17 cells produce IFN-γor IL-10 and are regulated by IL-1β[J].Nature,2012,484(7395):514.
[16]García JA,Volt H,Venegas C,et al.Disruption of the NF-κB/NLRP3 connection by melatonin requires retinoid-related orphan receptor-αand blocks the septic response in mice[J].Faseb Journal,2015,29(9):3863-3875.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |