CAJ | 학술논문

目的：探讨外源性硫化氢抑制氧化型低密度脂蛋白（ox-LDL）诱导内皮细胞组织因子（TF）表达的机制。方法分别用不同浓度NaHS（25、50、100和200μmol·L－1）和50 mg·L－1 ox-LDL孵育人脐静脉内皮细胞（HUVECs），用RT-PCR 和ELISA 法检测HUVECs TF mRNA 表达和蛋白含量，DCFH 法测细胞内活性氧（ROS）的含量，Western blot 测核蛋白转录因子（NFκB）的活化。结果 oxLDL 明显诱导HUVECs TF mRNA 的表达和蛋白含量增加；细胞内ROS 水平升高，NFκB 活化。NaHS 明显抑制了oxLDL 对TF mRNA 和蛋白表达的诱导作用，同时降低oxLDL 诱导的细胞内ROS生成和NFκB 活化。此外，NFκB 抑制剂BAY 117082（10μmol·L －1 ）或抗氧化剂N乙酰半胱氨酸（1 mmol·L －1 ）也能抑制oxLDL 诱导TF mRNA 表达和蛋白含量的增加，同时降低细胞内ROS 生成和NFκB 活化，该作用与200 μmol·L －1 NaHS 的效应相似。结论 NaHS 抑制oxLDL 诱导内皮细胞TF 的表达与降低ROS 产生和抑制NFκB 活化有关。
목적：탐토외원성류화경억제양화형저밀도지단백（ox-LDL）유도내피세포조직인자（TF）표체적궤제。방법분별용불동농도NaHS（25、50、100화200μmol·L－1）화50 mg·L－1 ox-LDL부육인제정맥내피세포（HUVECs），용RT-PCR 화ELISA 법검측HUVECs TF mRNA 표체화단백함량，DCFH 법측세포내활성양（ROS）적함량，Western blot 측핵단백전록인자（NFκB）적활화。결과 oxLDL 명현유도HUVECs TF mRNA 적표체화단백함량증가；세포내ROS 수평승고，NFκB 활화。NaHS 명현억제료oxLDL 대TF mRNA 화단백표체적유도작용，동시강저oxLDL 유도적세포내ROS생성화NFκB 활화。차외，NFκB 억제제BAY 117082（10μmol·L －1 ）혹항양화제N을선반광안산（1 mmol·L －1 ）야능억제oxLDL 유도TF mRNA 표체화단백함량적증가，동시강저세포내ROS 생성화NFκB 활화，해작용여200 μmol·L －1 NaHS 적효응상사。결론 NaHS 억제oxLDL 유도내피세포TF 적표체여강저ROS 산생화억제NFκB 활화유관。
Aim To investigate the mechanism for the inhibitory effect of hydrogen sulfide on the expression of tissue factor(TF)induced by oxidative low-density lipoprotein(ox-LDL)in endothelial cells.Methods Human umbilical vein endothelial cells (HUVECs ) were treated with 50 mg·L-1 ox-LDL in the absence or presence of different concentrations of NaHS (25 , 50,100 and 200 μmol·L-1 )for 24 h.The mRNA expression and protein content of TF in HUVECs were determined by reverse transcription PCR and ELISA, respectively.The content of intracellular reactive oxy-gen species (ROS)was determined by DCFH,an oxi-dative sensitive fluorescent indicator.The activation of nuclear factor-kappaB (NF-κB)was estimated by its expression in nuclear extracts analyzed by Western blot.Results Ox-LDL induced TF mRNA expression and increased TF protein content in HUVECs.The in-crease in intracellular ROS production and the activa-tion of NF-κB were observed in HUVECs treated with ox-LDL.However,NaHS could markedly inhibit the increases in TF mRNA and protein levels induced by ox-LDL.Also the elevation of intracellular ROS pro-duction and the activation of NF-κB elicited by ox-LDL were significantly suppressed by pretreatment with NaHS.In addition,pretreatment with BAY 1 1-7082 (10 μmol·L-1 ),the inhibitor of NF-κB or N-acetyl-L-cysteine(1 mmol·L-1 ),an antioxidant,could also decrease the TF mRNA and protein level as well as ROS production and NF-κB activation induced by ox-LDL in HUVECs,similar to the effects of 200 μmol· L-1 NaHS.Conclusion The mechanism for the in-hibitory effect of H2 S on the ox-LDL- induced TF ex-pression in endothelial cells may be related to inhibi-ting intracellular ROS production and subsequently NF-κB activation.