中国免疫学杂志
中國免疫學雜誌
중국면역학잡지
CHINESE JOURNAL OF IMMUNOLOGY
2015年
1期
45-47
,共3页
秦开秀%宗建春%方艳%王晓龙
秦開秀%宗建春%方豔%王曉龍
진개수%종건춘%방염%왕효룡
百草枯%肺泡上皮%凋亡%Elafin%核因子相关因子2
百草枯%肺泡上皮%凋亡%Elafin%覈因子相關因子2
백초고%폐포상피%조망%Elafin%핵인자상관인자2
Paraquat%A549%Apoptosis%Elafin%Nuclear factor erythroid like-2
目的:探讨转染Elafin对百草枯( Paraquat,PQ )诱导的肺泡上皮凋亡的影响及可能机制。方法:电穿孔法转染pEGFP-C1-Elafin进入A549细胞,培养24 h后,RT-PCR检测Elafin mRNA在A549细胞中的表达情况;并给予生理盐水、不同浓度PQ(5、50、100μmol/L)作用后,Annexin V/PI双染法流式细胞仪检测A549细胞凋亡率、活性氧类物质( Reactive oxygen species,ROS)含量。 Western blot 检测核因子相关因子2( Nuclear factor erythroid like-2, Nrf2)、血红素加氧酶-1( Heme oxygenase-1,HO-1)表达情况。结果:转入A549细胞Elafin成功表达。 PQ呈浓度依赖性导致A549细胞凋亡;PQ可使ROS含量明显增加,Nrf2、HO-1表达下调。 Elafin可抑制PQ诱导的细胞调亡,上调Nrf2、HO-1蛋白表达。结论:Elafin可以一定程度抑制PQ诱导的A549细胞凋亡,其机制可能与其上调抗氧化蛋白Nrf2等有关。
目的:探討轉染Elafin對百草枯( Paraquat,PQ )誘導的肺泡上皮凋亡的影響及可能機製。方法:電穿孔法轉染pEGFP-C1-Elafin進入A549細胞,培養24 h後,RT-PCR檢測Elafin mRNA在A549細胞中的錶達情況;併給予生理鹽水、不同濃度PQ(5、50、100μmol/L)作用後,Annexin V/PI雙染法流式細胞儀檢測A549細胞凋亡率、活性氧類物質( Reactive oxygen species,ROS)含量。 Western blot 檢測覈因子相關因子2( Nuclear factor erythroid like-2, Nrf2)、血紅素加氧酶-1( Heme oxygenase-1,HO-1)錶達情況。結果:轉入A549細胞Elafin成功錶達。 PQ呈濃度依賴性導緻A549細胞凋亡;PQ可使ROS含量明顯增加,Nrf2、HO-1錶達下調。 Elafin可抑製PQ誘導的細胞調亡,上調Nrf2、HO-1蛋白錶達。結論:Elafin可以一定程度抑製PQ誘導的A549細胞凋亡,其機製可能與其上調抗氧化蛋白Nrf2等有關。
목적:탐토전염Elafin대백초고( Paraquat,PQ )유도적폐포상피조망적영향급가능궤제。방법:전천공법전염pEGFP-C1-Elafin진입A549세포,배양24 h후,RT-PCR검측Elafin mRNA재A549세포중적표체정황;병급여생리염수、불동농도PQ(5、50、100μmol/L)작용후,Annexin V/PI쌍염법류식세포의검측A549세포조망솔、활성양류물질( Reactive oxygen species,ROS)함량。 Western blot 검측핵인자상관인자2( Nuclear factor erythroid like-2, Nrf2)、혈홍소가양매-1( Heme oxygenase-1,HO-1)표체정황。결과:전입A549세포Elafin성공표체。 PQ정농도의뢰성도치A549세포조망;PQ가사ROS함량명현증가,Nrf2、HO-1표체하조。 Elafin가억제PQ유도적세포조망,상조Nrf2、HO-1단백표체。결론:Elafin가이일정정도억제PQ유도적A549세포조망,기궤제가능여기상조항양화단백Nrf2등유관。
Objective:To investigate the effect of recombinant Elafin on A549 apoptosis induced by paraquat and the underlying mechanism.Methods:pEGFP-C1-Elafin was transformed into A549 competent cells by electroporation.Transformed A549 was cultured for additional 24 h before Elafin mRNA was detected by RT-PCR and cocultured with or without various concentration of paraquat( PQ ) for different duration.A549 apoptosis percentage and reactive oxygen species ( ROS ) content were tested by flow cytometry .Nuclear factor erythroid like-2(Nrf2) heme oxygenase-1(HO-1) were examed by Western blot .Results:Elafin expressed successfully in A549 after transformation.PQ caused A549 apoptosis in a concentration dependent manner and leaded to ROS content increasing and Nrf2 and HO-1 decreasing.However,elafin could inhibit ROS production and A549 apoptosis,upregulate Nrf2 and HO-1.Conclusion:Elafin could restrict A549 apoptosis to some extent and the possible mechanism lied in its ability to upregulate Nrf2 ex-pression.