中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2012年
5期
436-439
,共4页
纪伟宁%杨毅宁%马依彤%陈邦党%向阳%刘芬%李晓梅%陈铀%于子翔
紀偉寧%楊毅寧%馬依彤%陳邦黨%嚮暘%劉芬%李曉梅%陳鈾%于子翔
기위저%양의저%마의동%진방당%향양%류분%리효매%진유%우자상
重组9型腺相关病毒%CaMEK基因%缺血再灌注损伤%凋亡
重組9型腺相關病毒%CaMEK基因%缺血再灌註損傷%凋亡
중조9형선상관병독%CaMEK기인%결혈재관주손상%조망
Recombinant adeno-associated virus serotype 9%CaMEK gene%Ischemia reperfusioninduced injury%Apoptosis
目的 观察重组9型腺相关病毒(recombinant adeno-associated virus serotype 9,rAAV9)携带CaMEK基因转导心肌细胞是否能减轻缺血再灌注损伤诱导的心肌细胞凋亡.方法 分离培养SD大鼠心肌细胞,建立I/R模型,rAAV9-CBA-CaMEK转染心肌细胞.实验分组:(1)正常对照组；(2) I/R组；(3) I/R+rAAV9-CBA-CaMEK组；(4) I/R+rAAV9-CBA-CaMEK+PD98059组.Westem blot测定心肌细胞ERK1/2磷酸化水平及Caspase-3、Bax蛋白的表达.结果 分离培养的原代心肌细胞经鉴定结果阳性,Western blot分析显示rAAV9介导CaMEK基因转染心肌细胞P-ERK1/2的表达高峰时间在第5天,并可减轻缺血再灌注诱导的心肌细胞凋亡,而给予PD98059处理后,这种心肌细胞保护作用消失.结论 rAAV9介导的CaMEK基因转染可显著减轻缺血再灌注诱导的心肌细胞凋亡.
目的 觀察重組9型腺相關病毒(recombinant adeno-associated virus serotype 9,rAAV9)攜帶CaMEK基因轉導心肌細胞是否能減輕缺血再灌註損傷誘導的心肌細胞凋亡.方法 分離培養SD大鼠心肌細胞,建立I/R模型,rAAV9-CBA-CaMEK轉染心肌細胞.實驗分組:(1)正常對照組；(2) I/R組；(3) I/R+rAAV9-CBA-CaMEK組；(4) I/R+rAAV9-CBA-CaMEK+PD98059組.Westem blot測定心肌細胞ERK1/2燐痠化水平及Caspase-3、Bax蛋白的錶達.結果 分離培養的原代心肌細胞經鑒定結果暘性,Western blot分析顯示rAAV9介導CaMEK基因轉染心肌細胞P-ERK1/2的錶達高峰時間在第5天,併可減輕缺血再灌註誘導的心肌細胞凋亡,而給予PD98059處理後,這種心肌細胞保護作用消失.結論 rAAV9介導的CaMEK基因轉染可顯著減輕缺血再灌註誘導的心肌細胞凋亡.
목적 관찰중조9형선상관병독(recombinant adeno-associated virus serotype 9,rAAV9)휴대CaMEK기인전도심기세포시부능감경결혈재관주손상유도적심기세포조망.방법 분리배양SD대서심기세포,건립I/R모형,rAAV9-CBA-CaMEK전염심기세포.실험분조:(1)정상대조조；(2) I/R조；(3) I/R+rAAV9-CBA-CaMEK조；(4) I/R+rAAV9-CBA-CaMEK+PD98059조.Westem blot측정심기세포ERK1/2린산화수평급Caspase-3、Bax단백적표체.결과 분리배양적원대심기세포경감정결과양성,Western blot분석현시rAAV9개도CaMEK기인전염심기세포P-ERK1/2적표체고봉시간재제5천,병가감경결혈재관주유도적심기세포조망,이급여PD98059처리후,저충심기세포보호작용소실.결론 rAAV9개도적CaMEK기인전염가현저감경결혈재관주유도적심기세포조망.
Objective To determine whether recombinant adeno-associated virus serotyoe 9 with CaMEK gene transfected on myocardial cells leads to reduce I/R-induced apoptosis.Methods Establish an ischemic/reperfusion(I/R) model of myocardial cells in vitro and the cells were divided into four experimental groups:(1) control group;(2) I/R group;(3) I/R+rAAV9-CBA-CaMEK group;(4) I/R+rAAV9-CBACaMEK+PD98059 group,respectively.The P-ERK1/2 and the Caspase-3,Bax were quqntitated by Western blot.Results These data clearly demonstrate that AAV9-mediated CaMEK gene transfected lead to active ERK1/2 and reduce I/R-induced apoptosis notablely.Conclusion rAAV9-mediated CaMEK gene transfected on myocardial cells can reduce I/R-induced apoptosis.
