硝普钠诱导肝星状细胞HSC-T6凋亡及其机制  

Apoptosis of hepatic stellate cells (HSC-T6) induced by sodium nitroprusside and mechanisms involved

作　　者：张明明[1] 杨帅[2] 高福禄[3] 马洪骏[4] 邳克江[4] 崔东来[1] 张振科[1] 

机构地区：[1]河北医科大学第二医院消化内科,河北省石家庄市050000 [2]承德医学院附属医院肿瘤外科,河北省承德市067000 [3]河北师范大学,河北省石家庄市050017 [4]河北医科大学,河北省石家庄市050017

出　　处：《世界华人消化杂志》2010年第8期761-766,共6页World Chinese Journal of Digestology

摘　　要：目的:探讨一氧化氮(nitric oxide,NO)供体硝普钠(sodium nitroprusside,SNP)在诱导肝星状细胞(hepatic stellate cell,HSC-T6)凋亡中的作用及其机制.方法:应用流式细胞仪和Hoechst33258染色法检测HSC凋亡;激光扫描共聚焦显微镜检测荧光标记NF-κBp65的核转位;Real-timePCR方法检测基质金属蛋白酶抑制因子-1(tissue inhibitors of matrix metalloproteinase-1,TIMP-1)、Ⅰ型前胶原(ProcollagenⅠ)、抗凋亡蛋白基因(growth arrest and DNA damage-inducible protein,GADD45β)mRNA表达.结果:SNP组HSC凋亡率较对照组显著增加(20.78%±5.91%vs3.25%±1.26%,P=0.031),Hoechst33258染色法显示SNP组HSC细胞核呈致密浓染块状或颗粒状的荧光,提示细胞出现凋亡;SNP抑制TNF介导活化的HSCNF-κBp65核转位;随着其剂量不断增加,TIMP-1,ProcollagenⅠ,GADD45βmRNA表达随之减少(P<0.05).结论:SNP能诱导HSC凋亡,减少TIMP-1,ProcollagenⅠmRNA表达,其机制可能与通过抑制NF-κB活性,减少抗凋亡蛋白基因GADD45β mRNA表达有关.AIM：To investigate whether nitric oxide （NO） donor sodium nitroprusside （SNP） can induce the apoptosis of hepatic stellate cells （HSC-T6） and to explore potential mechanisms involved.METHODS：The apoptosis of HSC-T6 cells was determined by flow cytometry and Hoechst staining.The nuclear translocation of nuclear factor-κB （NF-κB） p65 was detected by laserscanning confocal microscopy.The expression of tissue inhibitor of matrix metalloproteinase-1 （TIMP-1）,type I procollagen （procollagen I）,and growth arrest and DNA damage-inducible protein （GADD45β） mRNAs was detected by real-time reverse transcription-polymerase chain reaction （RT-PCR）.RESULTS：The apoptosis rate was significantly higher in HSC-T6 cells treated with SNP than in control cells （20.78% ± 5.91% vs 3.25% ± 1.26%,P=0.031）.Apoptotic HSC-T6 cells showed dense nuclear staining or granular fluorescence after Hoechst staining.Tumor necrosis factor-α （TNF-α）-mediated nuclear translocation of NF-κB p65 was inhibited by SNP treatment.With the increase in SNP dose,the expression levels of TIMP-1,procollagen I and GADD45β mRNAs were reduced （all P 0.05）.CONCLUSION：SNP can induce the apoptosis of HSC-T6 cells and reduce the expression of TIMP-1 and procollagen I mRNAs perhaps by inhibiting NF-κB activity and reducing GADD45β mRNA expression.

关 键 词：硝普钠 核因子-ΚB 肝纤维化 

分 类 号：R575.2[医药卫生—消化系统]