硫化氢对吸烟诱导的肺动脉平滑肌细胞表型转化及自噬的影响  

The influence of hydrogen sulfide on cigarette smoke extract induced PASMC phenotype transition and autophagy

作　　者：米文君 陈亚红[1] 李敏霞[1] 廖艺璇 孙婉璐 贾国华 石丹丹[2] 齐永芬 MI Wen-jun;CHEN Ya-hong;LI Min-xia;LIAO Yi-xuan;SUN Wan-lu;JIA Guo-hua;SHI Dan-dan;QI Yong-fen(Department of Respiratory and Critical Care Medicine,Peking University Third Hospital,Beijing 100191,China;Medical Research Center,Peking University Third Hospital,Beijing 100191,China;Laboratory of Cardiovascular Bioactive Molecule,School of Basic Medical Sciences,Peking University Health Science Center,Beijing 100191,China)

机构地区：[1]北京大学第三医院呼吸与危重症医学科,北京100191 [2]北京大学第三医院中心实验室,北京100191 [3]北京大学医学部基础医学院生物活性小分子实验室,北京100191

出　　处：《中国医学前沿杂志（电子版）》2018年第9期36-42,共7页Chinese Journal of the Frontiers of Medical Science(Electronic Version)

基　　金：国家自然科学基金(81370141);国家重点研发计划"重大慢性非传染性疾病防控研究"专项课题(2016YFC1304301)

摘　　要：目的探究硫化氢(hydrogen sulfide,H2S)对吸烟诱导的肺动脉平滑肌细胞(pulmonary arterial smooth muscle cell,PASMC)表型转化及自噬的影响。方法在PASMC上,加入不同浓度(1%~10%)香烟烟雾提取物(cigarette smoke extract,CSE),用四甲基偶氮唑盐(methyl thiazolyl tetrazolium,MTT)法测定细胞增殖,用蛋白质印迹法测定增殖细胞核抗原(proliferating cell nuclear antigen,PCNA),测定平滑肌蛋白22α(smooth muscle protein 22α,SM22α)水平反映细胞表型转化,测定微管相关蛋白轻链3(microtubule-associated protein light chain 3,LC3)水平反映自噬变化;加入不同浓度(50~400μmol/L)的硫氢化钠(sodium hydrosulfide,Na HS),研究H2S对CSE诱导的细胞增殖、表型转化及自噬的影响。结果与对照组相比,5%CSE组细胞增殖增加了77.6%(P<0.01);与5%CSE组相比,10%CSE组细胞增殖下降了60.7%(P<0.01)。与对照组相比,5%CSE组PCNA表达增加了90.9%(P<0.05),SM22α表达下降了48.9%(P<0.05),5%CSE组LC3Ⅱ/LC3Ⅰ显著升高(P<0.05)。与对照组相比,5%CSE组、5%CSE+50μmol/L Na HS组、5%CSE+100μmol/L Na HS组、5%CSE+200μmol/L Na HS组PASMC增殖均显著增加(P_均<0.01)。与5%CSE组相比,5%CSE+100μmol/L Na HS组CSE诱导的PASMC增殖显著减少(P<0.05);5%CSE+400μmol/L Na HS组对CSE诱导的PASMC增殖抑制作用最强,较5%CSE组下降29.4%(P<0.01)。H2S对CSE诱导的PASMC PCNA表达增加具有抑制作用,5%CSE+200μmol/L Na HS组较5%CSE组抑制了40.9%(P<0.05);5%CSE+400μmol/L Na HS组H2S抑制PCNA表达增加的作用最显著,抑制了59.5%(P<0.01)。5%CSE+400μmol/L Na HS组SM22α表达较5%CSE组增加了53.4%(P<0.05);与对照组相比,5%CSE组PCNA表达显著增加(P<0.05),SM22α表达显著减少(P<0.05)。与5%CSE组相比,5%CSE+400μmol/L Na HS组CSE诱导的LC3Ⅱ/LC3Ⅰ升高抑制了45.7%(P<0.01);与对照组相比,5%CSE组LC3Ⅱ/LC3Ⅰ显著升高(P<0.05)。结论 H2S能够抑制CSE引起的PASMC增殖和表型转化,其作用机制可能与自噬相关。ObjectiveTo explore the infuence of hydrogen sulfde （H2S） on cigarette smoke extract （CSE） induced pulmonary artery smooth muscle cell （PASMC） phenotype transition and autophagy. MethodPASMCs were induced by di？erent concentrations （1% - 10%） of CSE. Cell proliferation was tested by methyl thiazolyl tetrazolium （MTT） assay, proliferating cell nuclear antigen （PCNA） was tested by Western blotting, and smooth muscle protein 22α （SM22α） re？ects cell phenotype transition, microtubule-associated protein light chain 3 （LC3） re？ects autophagy. Di？erent concentrations of H2S were added to explore the e？ect of H2S on CSE induced proliferation, phenotype switching and autophagy. ResultCompared with control group, PASMC in 5% CSE group increased 77.6% in cell proliferation （P〈0.01）; 10% CSE group reduced 60.7% compared with 5% CSE group （P〈0.01）. In 5% CSEgroup, PCNA expression increased 90.9% （P〈0.05）, SM22α expression decreased 48.9% （P〈0.05） and LC3Ⅱ/LC3Ⅰ increased dramatically compared with control group （P 〈 0.05）. The quantity of PASMC signi？cantly increased in 5% CSE group, 5% CSE＋50 μmol/L NaHS group, 5% CSE ＋ 100 μmol/L NaHS group and 5% CSE ＋200 μmol/L NaHS group compared with control group （Pall〈 0.01）. Compared with 5% CSE group, 5% CSE ＋ 100 μmol/L NaHS group CSE induced PASMC proliferation decreased signi？cantly （P〈0.05）; and the e？ect of 400 μmol/L NaHS group was the strongest （P 〈 0.01）. H2S inhibits the increase of CSE induced PASMC PCNA expression, the increase of PCNA expression induced by 5% CSE ＋ 200 μmol/L NaHS group was inhibited 40.9% than 5% CSE group （P 〈 0.05）, while the expression of PCNA was most signi？cantly inhibited by 5% CSE ＋ 400 μmol/LNaHS group （59.5%）; SM22α expression in 5% CSE ＋ 400 μmol/L NaHS group increased 53.4% compared with 5% CSE group （P 〈 0.05）. PCNA expressed signifcantly higher and SM22α expressed signi？cantly lower in 5% CSE grou

关 键 词：硫化氢 肺动脉平滑肌细胞 表型转化 自噬 

分 类 号：R544.1[医药卫生—心血管疾病]