红花注射液对肺缺血再灌注损伤时细胞凋亡及Bcl-2和Bax基因的影响
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浙江省卫生厅科研基金(SWS00021)资助


Effects of Safflower Injection on Pneumocyte Apoptosis and Bcl-2 and Bax Expression in Rabbits with Lung Ischemia-Reperfusion Injury
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    目的探讨细胞凋亡与肺缺血再灌注损伤的关系以及红花注射液的干预及机制。方法健康日本大耳白兔84只,随机分为对照组、缺血再灌注1、3、5h组和红花干预1、3、5h组。复制在体肺缺血再灌注损伤模型。采用电镜和原位缺口末端标记法观测肺组织细胞凋亡情况,并计算凋亡指数;免疫组织化学和原位杂交技术检测各组肺组织Bcl-2和Bax基因表达的变化。结果缺血再灌注组肺组织细胞凋亡指数和Bcl-2、Bax蛋白及mRNA均显著高于对照组(p<0.01)。红花干预组肺组织凋亡指数、Bax蛋白及Bax mRNA低于缺血再灌注组,而Bcl-2蛋白、Bcl-2mRNA以及Bcl-2与Bax的比值较缺血再灌注组上调(p<0.01或p<0.05)。电镜观察发现,缺血再灌注组肺毛细血管内皮细胞和Ⅱ型肺泡上皮细胞超微结构损伤明显,红花干预组损伤明显减轻。肺组织凋亡指数与Bax蛋白、Bax mRNA呈显著正相关(r分别=0.926,0.913;均p<0.01),与Bcl-2/Bax蛋白、Bcl-2/Bax mRNA的比值呈负相关(r分别=-0.367,-0.375;均p<0.01)。结论肺组织细胞凋亡参与了肺缺血再灌注损伤的发生,红花注射液可能通过下调Bax基因的表达,提高Bcl-2/Bax的比值抑制肺组织细胞凋亡,从而减轻肺缺血再灌注损伤。

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    Aim To explore the relationship between apoptosis in lung tissues and lung ischemia-reperfusion injury,and observe effects of safflower injection on pneumocyte apoptosis in lung ischemia-reperfusion injury.Methods Single lung in situ ischemia reperfusion animal model was used.84 rabbits were randomly divided into control group(C),ischemia/reperfusion 1 h group(IR1h),IR3h,IR5h,safflower injection 1 h group(SI1h),SI3h and SI5h.TdT mediated dUTP nick end labeling(TUNEL),electron microscope,immunocytochemistry and in situ hybridization(ISH) techniques were used to observe apoptosis,pulmonary ultrastructure and Bcl-2,Bax expression respectively in lung tissues of different group rabbits.Results Compared with control group,cell apoptosis,Bcl-2,Bax expression were up-regulated in lung tissues of IR groups(all of p<0.01).The safflower injection susppressed apoptosis as well as expression of Bax,but which improved the expression of Bcl-2 gene and the ratio of Bcl-2/Bax at the same time(p<0.01 or p<0.05,respectively).Electron microscope showed serious pulmonary ultrastructure damage in IR group,but no significant injury was found in SI group.There was a significant positive correlation between AI and Baxprotein,BaxmRNA(r=0.926,0.913;p<0.01,respectively),and there was a significant negative correlation between AI and the ratio of Bcl-2protein/Baxprotein,Bcl-2mRNA/BaxmRNA(r=-0.367,-0.375;p<0.01,respectively).Conclusion Apoptosis participates in lung ischemia reperfusion injury.The protective effects of safflower injection include suppressing the expression of Bax,blocking pneumocyte apoptosis and promoting Bcl-2 expression and the ratio of Bcl-2/Bax in lung ischemia-reperfusion injury.

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戴雍月,徐正衸,王万铁,邱晓晓,方周溪.红花注射液对肺缺血再灌注损伤时细胞凋亡及Bcl-2和Bax基因的影响[J].中国动脉硬化杂志,2007,15(6):415~418.

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  • 收稿日期:2006-12-26
  • 最后修改日期:2007-06-01
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