肾上腺素受体介导心脏纤维化模型中核转录因子NF-κB的活化  

Activation of transcription factor NF-κB in a rat model of cardiac fibrosis induced byβ-adrenoceptor stimulation

作　　者：殷倩[1,2] 鲁海燕[2,3] 杨承志[2] 田爱炬[2] 杨秋香[2,3] 张幼怡[2] 郑晓晖[3] 李子健[2] 郑小璞[1] 

机构地区：[1]西安交通大学医学院第一附属医院心内科,西安710061 [2]北京大学第三医院心内科血管医学研究所卫生部心血管分子生物学与调节肽重点实验室分子心血管学教育部重点实验室心血管受体研究北京市重点实验室,北京100191 [3]西北大学生命科学学院西部资源生物与现代生物技术教育部重点实验室,西安710069

出　　处：《北京大学学报（医学版）》2014年第6期889-893,共5页Journal of Peking University:Health Sciences

基　　金：国家自然科学基金(81471893,81270157,81070078);北京市自然科学基金(7102158);教育部长江学者和创新团队发展计划项目(IRT1174);陕西省重点科技创新团队计划项目(2013KCT-24)资助~~

摘　　要：目的：应用非选择性β肾上腺素受体激动剂——异丙基肾上腺素（isoproterenol,ISO）连续皮下注射7d诱导心脏纤维化模型,并观察此模型中炎症调控核转录因子NF-κB的变化.方法：采用10周龄体重为280 ～320 g的SD大鼠,ISO以0.25 mg/（kg·d）剂量连续背部皮下注射7d诱导心脏纤维化模型.心脏组织切片进行苦味酸-天狼猩红染色观察纤维化,以胶原容量分数进行胶原面积定量统计.应用real-time PCR法检测心脏组织中Ⅰ型/Ⅲ型胶原及炎症因子IL-6的mRNA表达.组织切片进行HE染色观察心脏病理变化.免疫组织化学方法检测心脏组织中NF-κB在细胞内的定位,Western blot法检测心脏组织中磷酸化的NF-κB水平.结果：ISO诱导的心脏纤维化模型中天狼猩红染色面积明显大于对照（control,CON）组,且ISO组的胶原容积分数显著高于CON组（12.01±1.64vs.0.95±0.06,P＜0.001）;ISO组Ⅰ、Ⅲ型胶原的mRNA表达显著高于CON组（Ⅰ型胶原：10.51±0.47vs.0.98±0.02,P＜0.001;Ⅲ型胶原：9.58±1.33 vs.1.02±0.02,P＜0.001）.ISO组心脏组织中可见细胞核增多、聚集,提示可能炎症细胞浸润,且炎症因子IL-6的mRNA表达水平明显增加（1.64±0.18 vs.1.04±0.07,P ＜0.01）.CON组NF-κB主要定位于心肌细胞胞浆中,心肌细胞核中未见NF-κB的定位;ISO组出现明显NF-κB核转位活化,心肌细胞胞浆中的NF-κB较CON组减少,心肌细胞核及间质细胞核均可见明显的NF-κB定位,同时,ISO组心脏组织磷酸化NF-κB的水平明显高于CON组（10.83±2.05vs.1.05±0.27,P＜0.001）.结论：连续7d皮下注射ISO可成功诱导心脏纤维化模型,且此模型心脏组织中活化的核转录因子NF-κB增加.Objective：To establish a model of cardiac fibrosis induced by isoproterenol （ISO）,the non-selective β adrenoceptor agonist,injected subcutaneously for 7 days in rats,and to observe changes of transcription factor NF-κB in the model.Methods：Male SD rats weighing 280-320 g were injected with ISO [0.25 mg/（kg · d）] subcutaneously for 7 days to induce cardiac fibrosis.The collagen volume fraction was determined by quantitative morphometry of picrosirius red stained left ventricular sections.Collagen types Ⅰ/Ⅲ and IL-6 mRNA expressions were analyzed by real time PCR.The pathological changes of the heart were investigated by Hematoxylin and Eosin staining.NF-κB was localized by immunohistochemistry （IHC） and phosphorylated NF-κB levels were assessed by Western blot analysis.Results：Compared with the controls,ISO significantly elevated the sirius red stained area and collagen volume fraction （12.01 ± 1.644 vs.0.95 ± 0.067,P ＜ 0.001）.Similarly,ISO increased the mRNA expressions of collagen Ⅰ and collagen Ⅲ of the heart compared with the controls （10.51 ± 0.47 vs.0.98 ±0.02,P＜0.001 for collagen Ⅰ; 9.58 ±1.33 vs.1.02 ±0.02,P＜0.001 for collagen Ⅲ）.The number of nuclei was increased and nuclear accumulation was presented in myocardial tissue induced by ISO.The mRNA expression of IL-6 increased in ISO group （1.64 ±0.18 vs.1.04 ±0.07,P ＜0.01）.ISO induced NF-κB nuclear translocation,accompanied by an increase in phosphorylation of NF-κB （10.83 ±2.05 vs.1.05 ±0.27,P ＜0.001）.Conclusion：We conclude that the model of cardiac fibrosis can be successfully induced by ISO injected subcutaneously for 7 days in rats and the activation of nuclear factor NF-κB increased by β-adrenoceptor stimulation.

关 键 词：心脏疾病 纤维化 异丙肾上腺素 NF-κB 

分 类 号：R542.23[医药卫生—心血管疾病]