中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (20): 3768-3772.doi: 10.3969/j.issn.1673-8225.2012.20.036

• 组织构建基础实验 basic experiments in tissue construction • 上一篇    下一篇

心外膜脂肪组织转染人apM1基因抑制兔冠状动脉粥样硬化的形成**☆

杨文凯1, 2，董念国1，陈  庆3，王现国1，谢  霆1，陈新忠1   

  1. 华中科技大学同济医学院附属协和医院，1心外科，3普外科，湖北省武汉市  430022；2江西省人民医院胸心外科，江西省南昌市  330006
  • 收稿日期:2012-01-19 修回日期:2012-02-28 出版日期:2012-05-13 发布日期:2012-05-13
  • 通讯作者: 陈新忠，博士，主治医师，华中科技大学同济医学院附属协和医院心外科，湖北省武汉市 430022 xinzhongchen@yahoo.com.cn
  • 作者简介:杨文凯☆，男，1975年生，山东省新泰市人，汉族，华中科技大学同济医学院在读博士，主治医师，主要从事心外科、体外循环基础和临床的研究。 doctor_wenkaiyang@yahoo.com.cn
  • 基金资助:

    国家自然科学基金项目(30872541)；博士点基金新教师项目资助课题(200804871116)。

Human apM1 gene transfers into the epicardial adipose tissue to inhibit the formation of coronary atherosclerosis in rabbits

Yang Wen-kai1, 2, Dong Nian-guo1, Chen Qing3, Wang Xian-guo1, Xie Ting1, Chen Xin-zhong1   

  1. 1Department of Cardiac Surgery, 3Department of General Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan  430022, Hubei Province, China; 2Department of Cardiac Surgery, Jiangxi Provincial People’s Hospital, Nanchang  330006, Jiangxi Province, China
  • Received:2012-01-19 Revised:2012-02-28 Online:2012-05-13 Published:2012-05-13
  • Contact: Chen Xin-zhong, Doctor, Attending physician, Department of Cardiac Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, Hubei Province, China xinzhongchen@yahoo.com.cn
  • About author:Yang Wen-kai☆, Studying for doctorate, Attending physician, Department of Cardiac Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, Hubei Province, China; Department of Cardiac Surgery, Jiangxi Provincial People’s Hospital, Nanchang 330006, Jiangxi Province, China doctor_wenkaiyang@yahoo.com.cn
  • Supported by:

    the National Natural Science Foundation of China, No. 30872541*; Doctoral Foundation for New Teachers, No. 200804871116*

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摘要：

背景：心外膜脂肪分泌的脂联素水平下降可能是导致冠状动脉粥样硬化的主要原因之一。
目的：观察人脂联素基因apM1转染至兔心外膜脂肪组织对高脂喂食兔冠状动脉粥样硬化形成的影响。
方法：向兔心包腔内注射脂质体包裹的pEGFP-apM1重组质粒50 μL，分别于注射后2，7，28 d观察转染效率。采用高脂喂食方法制备兔冠状动脉粥样硬化模型，并按上述方法转染apM1基因，于转染后4周取材。
结果与结论：心包腔内转染apM1基因2 d后即可检测到兔心外膜脂肪组织apM1基因高表达，并可持续至28 d。心外膜脂肪转染apM1基因对高脂喂食引起的外周血脂联素、肿瘤坏死因子α、总胆固醇、高密度脂蛋白胆固醇及低密度脂蛋白胆固醇水平无显著影响，但可使心包腔液中脂联素水平显著增加、肿瘤坏死因子α水平显著降低(P < 0.01)，冠状动脉内膜/中膜厚度比减小40.66%。说明经心包腔注射可将apM1基因有效转染入心外膜脂肪组织，并可在局部通过抑制高脂诱导的炎性因子释放而抑制冠状动脉粥样硬化的形成。

关键词: 脂联素, apM1, 心外膜脂肪组织, 冠状动脉粥样硬化

Abstract:

BACKGROUND: The decreased level of adiponectin secreted from the epicardial adipose tissue may be one of the main reasons leading to coronary atherosclerosis.
OBJECTIVE: To investigate the effect of human apM1 gene transferring into the epicardial adipose tissue on the formation of coronary atherosclerosis in high-cholesterol fed rabbits.
METHODS: 50 μL mixture of pEGFP-apM1 and liposome was injected into the rabbit pericardial cavity. The epicardial adipose tissue was harvested at 2, 7 and 28 days respectively after injection and was made into fast frozen sections. The transfection efficiency and expression of apM1 were detected by fluorescence microscopy and immunohistochemical method. Twenty-four male New Zealand White rabbits were randomly divided into four groups: normal control group, high-fat group, high-fat plus pEGFP injection group, high-fat plus pEGFP-apM1 injection group. At 4 weeks after injection, the expression of apM1 in epicardial adipose tissue was detected by Western blot analysis.
RESULTS AND CONCLUSION: In the rabbit epicardial adipose tissue, the high expression of apM1 was detected at 2 days after injection and until to 28 days. The effective transfection was not detected in the epicardial connective and myocardial tissues. The levels of adiponectin, tumor necrosis factor-α, total cholesterol, high density lipoprotein cholesterol and low density lipoprotein cholesterol in the peripheral blood changed insignificantly change after pEGFP-apM1 injection into the pericardial cavity. In high-fat plus pEGFP-apM1 injection group, the distinct increase of adiponectin and decrease of tumor necrosis factor-α (P < 0.01) were measured in the pericardial cavity fluid. Compared with the high-fat group, pEGFP-apM1 injection could decrease the intima/media ratio of left coronary artery to 40.66%. The apM1 gene could be effectively transferred into the epicardial adipose tissue through the pericardial cavity injection. The increase of adiponectin expression in the epicardial adipose tissue could inhibit local inflammatory cytokines release induced by high-fat feeding and inhibit the formation of coronary atherosclerosis.

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