石曌玲, 刘曼玲, 殷玥, 李晨, 杨峥, 薛涵, 王一石, 马恒. 脂质运载蛋白-2在川崎病相关心肌损伤中的作用及机制研究[J]. 心脏杂志, 2019, 30(1): 1-7. DOI: 10.12125/j.chj.201806035
    引用本文: 石曌玲, 刘曼玲, 殷玥, 李晨, 杨峥, 薛涵, 王一石, 马恒. 脂质运载蛋白-2在川崎病相关心肌损伤中的作用及机制研究[J]. 心脏杂志, 2019, 30(1): 1-7. DOI: 10.12125/j.chj.201806035
    Zhao-ling SHI, Man-ling LIU, Yue YIN, Chen LI, Zheng YANG, Han XUE, Yi-shi WANG, Heng MA. Role and mechanism of lipocalin-2 in Kawasaki disease-related myocardial injury[J]. Chinese Heart Journal, 2019, 30(1): 1-7. DOI: 10.12125/j.chj.201806035
    Citation: Zhao-ling SHI, Man-ling LIU, Yue YIN, Chen LI, Zheng YANG, Han XUE, Yi-shi WANG, Heng MA. Role and mechanism of lipocalin-2 in Kawasaki disease-related myocardial injury[J]. Chinese Heart Journal, 2019, 30(1): 1-7. DOI: 10.12125/j.chj.201806035

    脂质运载蛋白-2在川崎病相关心肌损伤中的作用及机制研究

    Role and mechanism of lipocalin-2 in Kawasaki disease-related myocardial injury

    • 摘要:
        目的   探讨川崎病(KD)相关心肌损伤的可能机制。
        方法   将45只(1月龄)SD大鼠随机分为对照组、干酪乳杆菌细胞壁成分(LCWE)模型组和4-苯基丁酸(4-PBA)治疗组。建立大鼠川崎病模型4周后,用酶联免疫吸附剂测定法ELISA检测KD大鼠血浆和心肌组织匀浆中脂质运载蛋白(Lcn)2的含量;超声心动图检测大鼠心脏功能;各组大鼠心肌组织制作石蜡切片并进行细胞凋亡TUNEL染色;蛋白质印迹法(Western blot)检测大鼠心肌内质网应激的标志物表达变化。离体实验采用原代培养新生大鼠心肌细胞,Lcn 2处理48 h后检测细胞活力、半胱氨酸天冬氨酸蛋白酶(Caspase)3活性以及内质网应激程度。
        结果   与对照组比较,KD大鼠血浆和心肌组织匀浆中Lcn 2的含量显著升高(P<0.05)。用Lcn 2(10 ng/ml)处理原代培养的心肌细胞48 h后可导致心肌细胞活力显著降低(P<0.05),同时标志物活化转录因子(ATF)6、CCAAT/增强子结合蛋白同源蛋白(CHOP)、磷酸化的蛋白激酶R样内质网激酶(p-PERK)、磷酸化的真核起始因子(p-elF)2α均出现升高,导致凋亡信号Caspase 12和活化的Caspase(cleave-Caspase)3水平显著增高(P<0.05)。KD大鼠心肌出现促凋亡性内质网应激,ATF6、CHOP、p-PERK、p-elF2α均显著升高,导致凋亡信号Caspase 12和cleave-Caspase 3水平显著增高(P<0.05)。同时伴有左室舒张末期内径(LVIDd)显著增加(P<0.05),左室射血分数(LVEF)显著降低(P<0.05)。使用4-PBA后能够有效抑制KD状态下的心肌内质网应激程度,降低ATF6、CHOP、p-PERK、p-elF2α水平,降低Caspase 12和cleave-Caspase 3水平(P<0.05),同时有效提升LVEF(P<0.05),改善KD大鼠心脏泵血功能,减少LVIDd(P<0.05),并最终改善KD大鼠舒张功能。
        结论   KD诱发心脏收缩舒张功能障碍同时伴有Lcn 2 显著升高。Lcn 2对心肌细胞的直接作用是促凋亡性内质网应激。抑制KD状态下的心肌内质网应激可抑制心肌凋亡改善心脏功能。

       

      Abstract:
        AIM   To investigate possible mechanisms of Kawasaki disease-related myocardial injury in a rat model of Kawasaki disease established utilizing Lactobacillus casei cell wall extract (LCWE).
        METHODS   Forty-five one-month old SD rats were randomly divided into control group, LCWE model group and 4-phenylbutyrate (4-PBA) treatment group. After 4 weeks of modeling, the content of Lcn 2 in the plasma and myocardial tissue homogenates of Kawasaki disease rats was detected by ELISA. The heart function of rats was detected by echocardiography. Paraffin sections were made from rat myocardial tissue and TUNEL staining was performed. Marker changes in expression of rat endoplasmic reticulum stress were detected by Western blotting. The primary culture of neonatal rat cardiomyocytes was performed in vitro. Cell viability, Caspase 3 activity, and endoplasmic reticulum stress were measured 48h after Lcn 2 treatment.
        RESULTS   The levels of Lcn 2 in Kawasaki disease rats were significantly higher than those in the control group (P<0.05). Lcn 2 induced apoptosis in cardiomyocytes. ER stress markers, ATF6, CHOP, p-PERK and p-elF2α in cardiomyocytes all increased, resulting in a significant increase in apoptotic signals Caspase 12 and cleave-Caspase 3 levels. Quantitative detection of Caspase 3 activity in cardiomyocytes demonstrated significant increases (P<0.05). Inhibition of endoplasmic reticulum stress by 4-PBA reduced the levels of ATF6, CHOP, p-PERK and p-elF2α and decreased the levels of Caspase 12 and cleave-Caspase 3 (P<0.05), which in turn relieved cardiomyocyte apoptosis and improved cardiac function in Kawasaki disease rats.
        CONCLUSION   Kawasaki disease induces systolic diastolic dysfunction and is accompanied by a significant increase in Lcn2. The direct effect of Lcn2 on cardiomyocytes is pro-apoptotic endoplasmic reticulum stress. Inhibition of endoplasmic reticulum stress in Kawasaki disease can inhibit myocardial apoptosis and improve cardiac function.

       

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