解剖学报 ›› 2020, Vol. 51 ›› Issue (3): 431-436.doi: 10.16098/j.issn.0529-1356.2020.03.020

• 组织学胚胎学发育生物学 • 上一篇    下一篇

过表达肝/骨/肾型碱性磷酸酶对小鼠急性心肌梗死后心室重塑的影响

 王立友 张冬颖*    

  1. 重庆医科大学附属第一医院心血管内科,重庆 400016
  • 收稿日期:2019-11-12 修回日期:2019-12-23 出版日期:2020-06-06 发布日期:2020-06-06
  • 通讯作者: 张冬颖 E-mail:cqmuzdy19@126.com
  • 基金资助:
    国家自然科学基金面上项目;中国心血管健康联盟心血管健康研究院“进阶”研究基金项目

Effect of alkaline phosphatase liver/bone/kidney overexpression on ventricular remodeling after acute myocardial infarction in mice#br#

WANG Li-you ZHANG Dong-ying*   

  1. Department of Cardiovascular Medicine, the First Affliated Hospital of Chongqing Medical University, Chongqing 400016, China
  • Received:2019-11-12 Revised:2019-12-23 Online:2020-06-06 Published:2020-06-06
  • Contact: ZHANG Dong-ying E-mail:cqmuzdy19@126.com
  • Supported by:
    National Natural Science Foundation of China;ACCESS Fund of China Heart House - China Cardiovascular Association

摘要:

目的 探讨肝/骨/肾型碱性磷酸酶(ALPL)过表达对小鼠急性心肌梗死(MI)后心室重塑的影响。  方法  36只8周龄雄性C57BL/6 J小鼠随机分为对照组(sham组),心肌梗死组(MI+Adv-EGFP组)和ALPL过表达组(MI+Adv-ALPL组),每组12只。术后2周,使用心脏彩色超声测定小鼠心脏功能,HE染色观察小鼠心脏组织病理变化,Real-time PCR检测小鼠心脏ALPL mRNA表达水平, Western blotting检测ALPL、α平滑肌肌动蛋白(α-SMA)、Ⅰ型胶原蛋白(ColⅠ)和Ⅲ型胶原蛋白(ColⅢ)表达情况,苦味酸天狼猩红染色偏振光法测定各组标本胶原容积分数(CVF)和Ⅰ型/Ⅲ型胶原比值。  结果  与MI组相比,过表达ALPL组小鼠心脏左室射血分数(LVEF)及左室短轴缩短率(LVFS)明显下降(P<0.05),过表达ALPL组HE染色可见心肌纤维断裂,排列紊乱,梗死区域纤维化明显,心脏组织α-SMA、ColⅠ、ColⅢ蛋白表达及CVF均显著增大(P<0.05),同时ColⅠ/ColⅢ比值显著增加(P<0.05)。  结论  ALPL过表达加重小鼠急性心肌梗死后心室病理性重塑。

关键词: 肝/骨/肾型碱性磷酸酶, 心肌梗死, 心室重塑Ⅰ型胶原, Ⅲ型胶原, 腺病毒转染, 实时定量聚合酶链反应, 免疫印迹法, 小鼠

Abstract:

Objective  To investigate the effect of alkaline phosphatase liver/bone/kidney (ALPL) overexpression on ventricular remodeling after acute myocardial infarction (MI).   Methods  Thirty six 8-week-old male C57BL/6 J mice were randomly divided into sham group(sham), MI group (MI+Adv-EGFP) and ALPL overexpression group (MI+Adv-ALPL) with 12 mice in each group. Two weeks after MI,cardiac function of mice was detected by echocardiography, pathological changes was detected by HE staining. ALPL mRNA expression in mouse heart was detected by Realtime PCR. The levels of ALPL, α-smooth muscle actin (α-SMA),collagen Ⅰ (ColⅠ) and collagen Ⅲ (ColⅢ) protein were detected by Western blotting. The collagen volume fraction (CVF) and the ratio of ColⅠ/ColⅢ were measured by polarizing method  with Sirius red staining.  Results  Compared with MI+Adv-EGFP group, the left ventricular ejection fraction (LVEF) (%) and left ventricular fractional shortening (LVFS) (%) of the heart in MI+Adv-ALPL group were reduced significantly (P<0.05). Hearts in the ALPL overexpression group showed myocardial fiber broken, and disordered and fibrous scars were obvious in myocardial infarction area. The expression of ALPL and the expression of α-SMA, ColⅠ and ColⅢ protein in MI+Adv-ALPL group increased significantly (P<0.05), and the proportion of ColⅠ/ColⅢ in MI+Adv-ALPL group increased significantly (P<0.05).Compared with MI+Adv-EGFP group, the CVF and the proportion of the ratio of ColⅠ/ColⅢ in the MI+Adv-ALPL group increased significantly (P<0.05).   Conclusion  ALPL overexpression can promote the ventricular pathological remodeling after acute myocardial infarction in mice.

Key words: Alkaline phosphatase liver/bone/kidney, Myocardial infarction, Ventricular remodeling, Type Ⅰ collagen, Type Ⅲ collagen, Adenovirus transfection, Real-time PCR, Western blotting, Mouse

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