硫化氢对离体大鼠胰腺腺泡细胞炎症因子表达的影响  

Effect of hydrogen sulfide on caerulein-induced inflammatory cytokines in rat pancreatic acinar cells

作　　者：付兰英[1] 饶春燕[1] 赵晓晏[1] 

机构地区：[1]第三军医大学新桥医院消化内科,重庆400037

出　　处：《第三军医大学学报》2013年第8期784-788,共5页Journal of Third Military Medical University

摘　　要：目的以雨蛙素刺激的大鼠胰腺腺泡细胞为体外急性胰腺炎(acute pancreatitis,AP)模型,研究硫化氢(H2S)对AP时胰腺腺泡细胞炎症因子表达的影响及机制。方法分离25只SD大鼠胰腺腺泡细胞,用雨蛙素(10-7mol/L)诱导体外AP模型,用不同浓度NaHS(5、10、20、40μmol/L)及不同时间(10、30、60 min)NaHS(5μmol/L)处理,Real-timePCR检测炎症因子mRNA水平。再用CSE抑制剂PAG处理,Real-time PCR检测炎症因子mRNA水平及CSE mRNA水平。进一步用NF-κB抑制剂BAY 11-7082处理后,Real-time PCR检测炎症因子mRNA水平,Western blot检测浆内IκBα及核内NF-κBp65的蛋白表达。结果当5μmol/L NaHS预处理60 min时,TNF-α水平(14.70±1.10)、IL-1β水平(14.42±0.83)显著低于雨蛙素组(P<0.05),IL-10水平(12.20±0.55)显著高于雨蛙素组(P<0.05);CSE抑制剂PAG预处理60 min后,TNF-α水平(37.38±0.89)、IL-1β水平(24.45±0.76)显著高于雨蛙素组(P<0.05),CSE水平(0.14±0.03)、IL-10水平(0.14±0.03)显著低于雨蛙素组(P<0.05);NF-κB抑制剂BAY 11-7082预处理30 min,TNF-α水平(9.17±0.96)、IL-1β水平(2.90±0.17)显著低于雨蛙素组(P<0.05),IL-10水平(40.00±1.00)显著高于雨蛙素组(P<0.05),并且胞浆内IκBα(0.90±0.01)的降解及细胞核内NF-κBp65(0.70±0.04)的蛋白表达明显低于雨蛙素组(P<0.05)。结论小剂量H2S供体NaHS在雨蛙素孵育的胰腺腺泡细胞中具有一定抗炎作用,且该作用是通过抑制NF-κB通路从而抑制腺泡细胞表达促炎因子实现的。Objective To study the effect of hydrogen sulfide（H2S） on caerulein-induced inflammatory cytokines in rat pancreatic acinar cells through NF-κB pathway.Methods Pancreatic acinar cells were isolated from 25 SD rats,and caerulein（10-7 mol/L） was used to induce acute pancreatitis（AP） in vitro.The pancreatic acinar cells were pre-incubated with NaHS（5,10,20 and 40 μmol/L） for 30 min,NaHS（5 μmol/L） for different time（10,30 and 60 min）,cystathionine-γ-lyase（CSE） inhibitor of PAG（3 μmol/L） for 60 min,and NF-κB inhibitor of BAY 11-7082 for 30 min plus NaHS（5 μmol/L） for another 60 min,respectively,and then were treated with caerulein for 60 min.The cells treated with caerulein alone were served as control.The mRNA levels of inflammatory cytokines and CSE were measured by real-time PCR,and the levels of IκBα and NF-κBp65 were measured by Western blotting.Results As compared with the caerulein group,the levels of TNF-α（14.70±1.10） and IL-1β（14.42±0.83） were significantly decreased and IL-10（12.20±0.55） were significantly increased in cells pretreated with 5 μmol/L of NaHS for 60 min（P0.05）,while the levels of TNF-α（37.38±0.89） and IL-1β（24.45±0.76） were significantly increased and the level of CSE（0.14±0.03） and IL-10（0.14±0.03） were significantly decreased in cells pretreated with PAG for 60 min（P0.05）.In the cells pretreated with BAY 11-7082 for 30 min,the levels of TNF-α（9.17±0.96）,IL-1β（2.90±0.17） and nuclear NF-κBp65（0.70±0.04） were significantly decreased,but the level of IL-10（40.00±1.00） and IκBα（0.90±0.01） were increased as compared with the caerulein group（P0.05）.Conclusion A low dose of NaHS can inhibit the caerulein-induced expression of inflammatory cytokines and increase anti-inflammatory cytokines through suppressing the activation of NF-κB pathway,suggesting the low dose of H2S plays an anti-inflammatory role in AP.

关 键 词：H2S 胰腺腺泡细胞 NF-ΚB 细胞活素 

分 类 号：R576[医药卫生—消化系统] R914.3[医药卫生—内科学]