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中国人民解放军总医院老年心血管病研究所
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中华老年多器官疾病杂志编辑委员会
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创刊人 王士雯
总编辑 范利
副总编辑 陈韵岱
执行主编 叶大训
编辑部主任 王雪萍
ISSN 1671-5403
CN 11-4786
创刊时间 2002年
出版周期 月刊
邮发代号 82-408
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段明霞,唐其柱.还原型烟酰胺腺嘌呤二核苷酸磷酸氧化酶在脂多糖诱导的小鼠急性心肌损伤组织中的表达及意义[J].中华老年多器官疾病杂志,2018,17(2):133~138
还原型烟酰胺腺嘌呤二核苷酸磷酸氧化酶在脂多糖诱导的小鼠急性心肌损伤组织中的表达及意义
Expression of reduced nicotinamide adenine dinucleotide phosphate oxidase in acute myocardial injury mice induced by lipopolysaccharide and its significance
投稿时间:2017-09-27  修订日期:2017-11-06
DOI:10.11915/j.issn.1671-5403.2018.02.029
中文关键词:  脂多糖类;细胞凋亡;氧化性应激;烟酰胺腺嘌呤二核苷酸磷酸;心肌损伤
英文关键词:lipopolysaccharides; apoptosis;oxidative stress; nicotinamide adenine dinucleotide phosphate; myocardial damage
基金项目:国家自然科学基金(81470516,2)
作者单位E-mail
段明霞 武汉大学人民医院心内科,武汉大学心血管病研究所,心血管病湖北省重点实验室, 武汉 430060  
唐其柱 武汉大学人民医院心内科,武汉大学心血管病研究所,心血管病湖北省重点实验室, 武汉 430060 qztang@whu.edu.cn 
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中文摘要:
      目的 探讨还原型烟酰胺腺嘌呤二核苷酸磷酸(NADPH)氧化酶的非吞噬细胞氧化酶(Nox) 家族在脂多糖(LPS)诱导的小鼠急性心肌损伤组织中的表达及意义。方法 选取8~10周龄无特定病原体C57/BL6雄性小鼠30只,随机分为对照组和LPS组,每组15只。LPS组小鼠通过腹腔注射大剂量LPS(10 mg/kg)诱导急性心肌损伤模型,对照组腹腔注射等量生理盐水,观察6 h后取材,逆转录-聚合酶链反应(RT-PCR)检测Nox、B细胞淋巴瘤/白血病蛋白-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、半胱氨酸天门冬氨酸特异性蛋白酶3(Caspase 3)基因表达,Western blotting检测Nox 2、Nox 4、Bax、Bcl-2和Caspase 3蛋白表达,免疫组织化学法检测4羟基壬烯醛(4-HNE)表达,末端脱氧核糖核酸转移酶(TdT)介导的脱氧尿苷三磷酸(dUTP)切口末端标记技术(TUNEL)法检测心肌组织细胞凋亡,比较两组结果差异。使用SPSS 13.0统计软件对数据进行分析,两组比较采用t检验。结果 相比对照组,LPS组小鼠心肌Nox 2、Nox 4、Bax基因和蛋白以及Caspase 3蛋白表达水平明显升高,Bcl-2基因和蛋白表达水平明显降低,差异具有统计学意义(P<0.05)。 免疫组织化学检测结果表明相比对照组,LPS组心肌组织4-HNE表达明显增加,差异具有统计学意义(P<0.05)。TUNEL检测结果表明相比对照组,LPS组心肌组织心肌凋亡细胞明显增加,差异具有统计学意义(P<0.05)。结论 NADPH氧化酶通过调控氧化应激和细胞凋亡参与急性心肌损伤的发生发展。
英文摘要:
      Objective To investigate the expression of non-phagocytic cell oxidase (Nox), reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, in mice of lipopolysaccharide (LPS)-induced acute myocardial injury and its significance. MethodsSPF male C57/BL6 mice (8-10 weeks old) were randomly assigned into control group (normal saline,n=15) and LPS group (n=15). LPS group was injected with large dose of LPS (10 mg/kg) to induce acute myocardial injury model, and the control group was only injected with saline equivalent. The mice were sacrificed in 6 h later, and the heart tissues were harvested. The mRNA and protein levels of Nox 2, Nox 4, Bcl-2-associated X protein (Bax), B cell lymphoma/leukemia-2 (Bcl-2) and cysteine aspastic acid-specific protease 3 (Caspase 3) were detected by RT-PCR and Western blotting respectively. The expression of lipid peroxidation products (4-HNE) in the myocardium was measured by immunohistochemistry. TUNEL was used to detect myocardial apoptosis. SPSS statistics 13.0 was used to perform the statistical analysis, and student’s t test was employed for comparison between groups. ResultsCompared with the control group, the mRNA and protein levels of Nox 2, Nox 4 and Bax were significantly increased, while those of Bcl-2 were decreased, and the protein expression of Caspase 3 was elevated in the LPS group (P<0.05). Immunohistochemical assay showed that the expression of 4-HNE was significantly higher in the mice of the LPS group (P<0.05). TUNEL assay indicated that there were more apoptotic cells in the LPS group (P<0.05). Conclusion NADPH oxidase takes part in the occurrence and development of acute myocardial injury through regulating oxidative stress and cell apoptosis.
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