Protective effect of Barbaloin in a rat model of myocardial ischemia reperfusion injury through the regulation of the CNPY2-PERK pathway
Protective effect of Barbaloin in a rat model of myocardial ischemia reperfusion injury through the regulation of the CNPY2-PERK pathway
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DOI：
10.3892/ijmm.2019.4123
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发表时间：
2019-05-01
影响因子：
5.4
通讯作者：
Liu, Gang
中科院分区：
文献类型：
--
作者：
Cui, Yue;Wang, Yongqiang;Liu, Gang
Barbaloin （Bar）具有心肌保护作用，但其作用机制尚不明确。内质网应激（ERS）介导的细胞凋亡途径在心肌缺血再灌注损伤（MIRI）的发病机制中起重要作用。抑制ERS可能显著改善MIRI的进展，并起到预防MIRI的作用。因此，基于目前对ers介导的心肌细胞凋亡和Bar的心脏保护作用的了解，本研究的目的是进一步评估Bar预处理在MIRI中的心肌保护作用和可能的机制。本研究建立MIR大鼠模型，随机分为4组。大鼠在心肌缺血前给予Bar (20 mg/kg)，每天1次，连用1周。测定心肌血清乳酸脱氢酶和肌酸激酶。采用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记法评价Bar预处理对MIRI的心肌保护作用。为了评估ERS信号通路是否参与Bar预处理的心肌保护机制，采用western blot分析、免疫组织化学或逆转录定量聚合酶链反应检测ERS相关蛋白、蛋白冠同源物2 （CNPY2）、葡萄糖调节蛋白78、转录激活因子4、C/ ebp同源蛋白（CHOP）、PKR内质网激酶（PERK）、caspase-12和caspase-3的表达水平。结果证实，Bar预处理可显著降低MIR引起的损伤和细胞凋亡水平。Bar预处理显著抑制心肌细胞内ers相关蛋白的表达。此外，免疫组化结果显示，Bar预处理显著抑制心肌细胞cnpy2阳性细胞凋亡率。因此，本研究结果提示CNPY2存在于心肌细胞中，并通过启动PERK-CHOP信号通路参与MIRI的发展。Bar预处理可能通过抑制CNPY2-PERK凋亡通路来减弱MIRI。
Barbaloin (Bar) has a myocardial protective effect, but its mechanism of action is uncertain. The endoplasmic reticulum stress (ERS)-mediated apoptosis pathway serves an important role in the pathogenesis of myocardial ischemia-reperfusion injury (MIRI). Inhibiting ERS may significantly improve the progression of MIRI and serve a role in its prevention. Therefore, based on current knowledge of ERS-mediated cardiomyocyte apoptosis and the cardioprotective effect of Bar, the purpose of the present study was to further evaluate the myocardial protective effect and potential mechanisms of Bar pretreatment in MIRI. The present study established a MIR rat model and randomly divided these rats into four groups. Prior to myocardial ischemia, Bar (20 mg/kg) was administered to rats once daily for 1 week. Myocardial blood serum lactate dehydrogenase and creatine kinase were subsequently measured. A terminal deoxynucleotidyl transferase mediated dUTP nick end labeling assay was used to evaluate the myocardial protective effect of Bar pretreatment on MIRI. To assess whether the ERS signaling pathway was involved in the myocardial protection mechanism of Bar pretreatment, the expression levels of ERS-associated proteins, protein canopy homolog 2 (CNPY2), glucose regulatory protein 78, transcriptional activator 4, C/EBP-homologous protein (CHOP), PKR endoplasmic reticulum kinase (PERK), caspase-12 and caspase-3 were detected by western blot analysis, immunohistochemistry or reverse transcription-quantitative polymerase chain reaction. The results confirmed that Bar pretreatment significantly reduced the damage and the level of apoptosis caused by MIR. Bar pretreatment significantly inhibited the expression of ERS-associated proteins in cardiomyocytes. In addition, the immunohistochemistry results demonstrated that Bar pretreatment significantly inhibited the CNPY2-positive cell apoptosis ratio of cardiomyocytes. Therefore, the results of the current study suggested that CNPY2 is present in cardiomyocytes and participates in the development of MIRI by initiating the PERK-CHOP signaling pathway. Bar pretreatment may attenuate MIRI by inhibiting the CNPY2-PERK apoptotic pathway.