CAJ | 학술논문

目的研究短期缺氧-复氧对大鼠血管平滑肌细胞myocardin基因表达的影响.方法无菌条件下留取大鼠胸主动脉中膜,采用组织贴块法培养的第4～7代细胞,通过观察细胞在缺氧环境下形态学变化、死亡细胞计数等,判断缺氧刺激对体外培养平滑肌细胞活性的影响,并与正常培养箱内对照进行比较.提取细胞总RNA,用RT-PCR法,研究平滑肌细胞在缺氧及复氧后不同时间段myocardin mRNA的表达变化.结果培养细胞呈典型的"峰谷"样生长.自制缺氧装置24 h内氧分压明显降低,且pH值波动不明显.RT-PCR结果表明,与正常对照组相比,平滑肌细胞myocardin mRNA表达在缺氧12、24 h后逐渐降低[(0.871±0.053)与(0.573±0.074)、(1.329±0.042),P＜0.05].缺氧12 h降低最明显.且随着缺氧时间的延长,myocardin基因的表达水平逐渐回升,并增高;缺氧24 h再复氧6 h后,其表达水平逐渐回升,复氧12 h接近正常表达水平.结论短期缺氧-复氧影响大鼠血管平滑肌细胞myocardin基因的表达.
목적 연구단기결양-복양대대서혈관평활기세포myocardin기인표체적영향.방법 무균조건하류취대서흉주동맥중막,채용조직첩괴법배양적제4～7대세포,통과관찰세포재결양배경하형태학변화、사망세포계수등,판단결양자격대체외배양평활기세포활성적영향,병여정상배양상내대조진행비교.제취세포총RNA,용RT-PCR법,연구평활기세포재결양급복양후불동시간단myocardin mRNA적표체변화.결과배양세포정전형적"봉곡"양생장.자제결양장치24 h내양분압명현강저,차pH치파동불명현.RT-PCR결과표명,여정상대조조상비,평활기세포myocardin mRNA표체재결양12、24 h후축점강저[(0.871±0.053)여(0.573±0.074)、(1.329±0.042),P＜0.05].결양12 h강저최명현.차수착결양시간적연장,myocardin기인적표체수평축점회승,병증고;결양24 h재복양6 h후,기표체수평축점회승,복양12 h접근정상표체수평.결론 단기결양-복양영향대서혈관평활기세포myocardin기인적표체.
Objective To investigate the influence of hypoxia-reoxygenation on the myocardin gene expression of cultured rats' vascular smooth muscle cells(SMC).Methods The SMC were isolated from the media of the thoracic aorta vessel of Sprague-Dawley rats,and cultured with attachment-block manner.The morphology and cell counting of the cultured cells under hypoxia conditions were observed and comparedto that under normal culture condition.Total RNA extracted from the cultured cells,the expression of myocardin mRNA in SMC were measured at hypoxia status and at various time after reoxygenation through RT-PCR.Results The rats' vessel SMC was successfully cultured and showed a "peak-valley" shape.Self-made hypoxia equipment can produce a lower oxygen partial pressure without significant variation of pH value which met the experiment requirements in 2 4 hours .However,in the hypoxia conditions,the expression level of myocardin was lowest at the 12th hours,then increased.After 24 hours of hypoxia,the expression levels of myocardin began to increase at the 6th hour of reoxygenation and reached a normal level at the 12th hour of reoxygenation.Conclusions Hypoxia-reoxygenation has an effect on the expression of myodardin gene.