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

北京大学学报（医学版） ›› 2014, Vol. 46 ›› Issue (6): 889-893.

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

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△

(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)

出版日期:2014-12-18 发布日期:2014-12-18

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

YIN Qian1,2, LU Hai-yan2,3, YANG Cheng-zhi2, TIAN Ai-ju2, YANG Qiu-xiang2，3, ZHANG You-yi2, ZHENG Xiao-hui3△, LI Zi-jian2△, ZHENG Xiao-pu1△

(1.Department of Cardiovascular Medicine, First Affiliated Hospital of Medical College of Xi’an Jiaotong University, Xi’an 710061, China; 2.Institute of Vascular Medicine, Peking University Third Hospital, Key Laboratory of Cardiovascular Molecular Biology and Regulatory Peptides, Ministry of Health, Key Laboratory of Molecular Cardiovascular Sciences, Ministry of Education and Beijing Key Laboratory of Cardiovascular Receptors Research， Beijing 100191， China； 3.Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, College of Life Sciences, Northwest University, Xi’an 710069, China)

Online:2014-12-18 Published:2014-12-18

摘要/Abstract

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7 d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7 d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±1.64 vs. 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±0.47 vs. 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±2.05 vs.1.05±0.27, P＜0.001)。结论：连续7 d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。

关键词: 心脏疾病, 纤维化, 异丙肾上腺素, NF-&kappa, B

Abstract: 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 Ⅰand 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.

Key words: Cardiomyopathies, Fibrosis, Isoproterenol, NF-kappaB

殷倩, 鲁海燕, 杨承志, 田爱炬, 杨秋香, 张幼怡, 郑晓晖, 李子健, 郑小璞. 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[J]. 北京大学学报（医学版）, 2014, 46(6): 889-893.

YIN Qian, LU Hai-Yan, YANG Cheng-Zhi, TIAN Ai-Ju, YANG Qiu-Xiang, ZHANG You-Yi, ZHENG Xiao-Hui, LI Zi-Jian, ZHENG Xiao-Pu. Activation of transcription factor NF-κB in a rat model of cardiac fibrosis induced by β-adrenoceptor stimulation[J]. Journal of Peking University(Health Sciences), 2014, 46(6): 889-893.

参考文献

相关文章 15

[1]	朱小玲,李文静,王宪娥,宋文莉,徐莉,张立,冯向辉,路瑞芳,释栋,孟焕新. 细胞色素B-245α链及胆固醇酯转运蛋白基因多态性与广泛型侵袭性牙周炎易感性的关系[J]. 北京大学学报（医学版）, 2022, 54(1): 18-22.
[2]	范莹莹,刘云,曹烨,谢秋菲. 海马参与雌激素加重咬合干扰致去卵巢大鼠慢性咬肌痛敏[J]. 北京大学学报（医学版）, 2022, 54(1): 40-47.
[3]	杜文瑜,杨静文,姜婷. 甲磺酸去铁胺促进大鼠颅骨临界骨缺损血管化骨再生的早期连续观察[J]. 北京大学学报（医学版）, 2021, 53(6): 1171-1177.
[4]	庞泳,张沙,杨华,周柔丽. LAPTM4B-35蛋白作为肝癌血清学诊断新标志物的探讨[J]. 北京大学学报（医学版）, 2021, 53(4): 710-715.
[5]	马向波,张学武,贾汝琳,高颖,刘洪江,刘玉芳,李英妮. 外周血淋巴细胞亚群检测在系统性硬化症治疗中的应用[J]. 北京大学学报（医学版）, 2021, 53(4): 721-727.
[6]	冯菁楠,高乐,孙一鑫,杨继春,邓思危,孙凤,詹思延. Xpert^®MTB/RIF对我国人群活动性肺结核和利福平耐药肺结核诊断准确性的meta分析[J]. 北京大学学报（医学版）, 2021, 53(2): 320-326.
[7]	刘世博,高辉,冯元春,李静,张彤,万利,刘燕鹰,李胜光,罗成华,张学武. 腹膜后纤维化致肾盂积水的临床分析:附17例报道[J]. 北京大学学报（医学版）, 2020, 52(6): 1069-1074.
[8]	赵世明,杨铁军,许春苗,郭孝峰,马永康,陈学军,李祥,何朝宏. 3.0T磁共振成像在接受过经尿道膀胱肿瘤切除术膀胱癌中诊断肌层浸润的应用[J]. 北京大学学报（医学版）, 2020, 52(4): 701-704.
[9]	轩艳,蔡宇,王啸轩,石巧,邱立新,栾庆先. 牙龈卟啉单胞菌感染对载脂蛋白e基因敲除小鼠动脉粥样硬化的影响[J]. 北京大学学报（医学版）, 2020, 52(4): 743-749.
[10]	陈章健,韩硕,郑湃,周淑佩,贾光. 纳米二氧化钛与葡萄糖亚慢性联合经口暴露对幼年大鼠血清叶酸和维生素B₁₂水平的影响[J]. 北京大学学报（医学版）, 2020, 52(3): 451-456.
[11]	赵磊,张洪宪,侯小飞,刘磊,陆敏,韩永,马潞林. 来氟米特联合羟苯磺酸钙治疗肾移植术后BK病毒相关性肾病[J]. 北京大学学报（医学版）, 2020, 52(2): 385-389.
[12]	马博,田志华,曲莉,刘月香,张宏,丁慧荣. 索拉菲尼诱导的肝癌耐药细胞株的建立及基因表达分析[J]. 北京大学学报（医学版）, 2020, 52(2): 207-213.
[13]	白向松,吕珑薇,周永胜. Tribbles同源蛋白3抑制人脂肪间充质干细胞成脂向分化[J]. 北京大学学报（医学版）, 2020, 52(1): 1-9.
[14]	田艳,朱军. p53 rs1625895基因多态性与弥漫大B细胞淋巴瘤预后相关性分析[J]. 北京大学学报（医学版）, 2019, 51(5): 791-796.
[15]	朱学华,杨明钰,夏海缀,何为,张智荧,刘余庆,肖春雷,马潞林,卢剑. 机器学习模型在预测肾结石输尿管软镜碎石术后早期结石清除率中的应用[J]. 北京大学学报（医学版）, 2019, 51(4): 653-659.

Metrics

Viewed

Full text

Abstract

Cited

Shared

Discussed