Heat Shock Protein 70 Protects the Heart from Ischemia/Reperfusion Injury through Inhibition of p38 MAPK Signaling
Heat Shock Protein 70 Protects the Heart from Ischemia/Reperfusion Injury through Inhibition of p38 MAPK Signaling
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热休克蛋白 70 通过抑制 p38 MAPK 信号传导保护心脏免受缺血/再灌注损伤
DOI：
10.1155/2020/3908641
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发表时间：
2020-03-21
影响因子：
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通讯作者：
Ji, Fu-hai
中科院分区：
文献类型：
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作者：
Song, Nan;Ma, Jiao;Ji, Fu-hai
热休克蛋白70（Hsp 70）具有心脏保护作用。p38丝裂原活化蛋白激酶（p38 MAPK）激活引起的细胞内钙超载是心肌缺血再灌注（I/R）损伤的重要原因。然而，Hsp 70是否与p38 MAPK信号转导相互作用尚不清楚。因此，本研究探讨了热休克蛋白70对p38 MAPK在缺血再灌注心肌损伤中的调节作用。方法采用新生大鼠心肌细胞缺氧缺糖6 h再复氧2 h（OGD/R），结扎左前动脉30 min再灌注30 min。在OGD/R或I/R之前使用p38 MAPK抑制剂（SB 203580）、Hsp 70抑制剂（Quercetin）和Hsp 70短发夹RNA（shRNA）。细胞存活率、乳酸脱氢酶（LDH）释放、血清心肌肌钙蛋白I（cTnI）、[Ca 2 +]i水平、细胞凋亡、心肌梗死面积、IL-1β和IL-6 mRNA水平、Hsp 70蛋白表达、磷酸化p38 MAPK（p-p38 MAPK）、肌浆网/内质网Ca 2 +-ATP酶2（SERCA 2）、磷酸化信号转导和转录激活因子3（p-STAT 3），和切割的半胱天冬酶3。结果p38 MAPK抑制剂SB 203580预处理可明显减轻OGD/R诱导的细胞损伤或I/R诱导的心肌损伤，表现为细胞活力增加，LDH释放减少，血清cTnI水平降低，心肌梗死面积缩小，[Ca 2 +]i超载减轻，细胞凋亡减轻，IL-1β和IL-6水平降低，p-p38 MAPK蛋白表达上调。SERCA 2、p-STAT 3和裂解型半胱天冬酶3。通过shRNA敲低Hsp 70加剧了OGD/R诱导的细胞损伤，SB 203580有效地消除了这种损伤。此外，槲皮素抑制Hsp 70增强I/R诱导的心肌损伤，而SB 203580预处理逆转槲皮素引起的有害影响。结论Hsp 70通过调节p38 MAPK信号通路抑制心肌I/R损伤时[Ca 2 +]i超载、炎症反应和细胞凋亡，为心肌保护策略提供新的思路。
Background Heat shock protein 70 (Hsp70) has been shown to exert cardioprotection. Intracellular calcium ([Ca2+]i) overload induced by p38 mitogen-activated protein kinase (p38 MAPK) activation contributes to cardiac ischemia/reperfusion (I/R) injury. However, whether Hsp70 interacts with p38 MAPK signaling is unclear. Therefore, this study investigated the regulation of p38 MAPK by Hsp70 in I/R-induced cardiac injury. Methods Neonatal rat cardiomyocytes were subjected to oxygen-glucose deprivation for 6 h followed by 2 h reoxygenation (OGD/R), and rats underwent left anterior artery ligation for 30 min followed by 30 min of reperfusion. The p38 MAPK inhibitor (SB203580), Hsp70 inhibitor (Quercetin), and Hsp70 short hairpin RNA (shRNA) were used prior to OGD/R or I/R. Cell viability, lactate dehydrogenase (LDH) release, serum cardiac troponin I (cTnI), [Ca2+]i levels, cell apoptosis, myocardial infarct size, mRNA level of IL-1β and IL-6, and protein expression of Hsp70, phosphorylated p38 MAPK (p-p38 MAPK), sarcoplasmic/endoplasmic reticulum Ca2+-ATPase2 (SERCA2), phosphorylated signal transducer and activator of transcription3 (p-STAT3), and cleaved caspase3 were assessed. Results Pretreatment with a p38 MAPK inhibitor, SB203580, significantly attenuated OGD/R-induced cell injury or I/R-induced myocardial injury, as evidenced by improved cell viability and lower LDH release, resulted in lower serum cTnI and myocardial infarct size, alleviation of [Ca2+]i overload and cell apoptosis, inhibition of IL-1β and IL-6, and modulation of protein expressions of p-p38 MAPK, SERCA2, p-STAT3, and cleaved-caspase3. Knockdown of Hsp70 by shRNA exacerbated OGD/R-induced cell injury, which was effectively abolished by SB203580. Moreover, inhibition of Hsp70 by quercetin enhanced I/R-induced myocardial injury, while SB203580 pretreatment reversed the harmful effects caused by quercetin. Conclusions Inhibition of Hsp70 aggravates [Ca2+]i overload, inflammation, and apoptosis through regulating p38 MAPK signaling during cardiac I/R injury, which may help provide novel insight into cardioprotective strategies.