Yuan Chunju, Chen Zhongpu, Zhou Qianxing
Department of Cardiovascular Medicine, Zhongda Hospital Southeast University, Nanjing, Jiangsu 210009, P.R. China.
Mol Med Rep. 2023 Jan;27(1). doi: 10.3892/mmr.2022.12907. Epub 2022 Dec 9.
Myocardial infarction (MI) refers to myocardial ischemic necrosis that is caused by coronary artery disease. Notably, crocin has protective effects on the heart. The present study aimed to i) investigate the protective effect of crocin, an active ingredient in Ellis and L., on myocardial ischemia and ii) to verify the interaction between crocin and kelch repeat and BTB domain‑containing protein 7 (KBTBD7), which is a novel member of the BTB‑kelch protein family. In the present study, the left anterior descending coronary artery was ligated to establish a myocardial ischemia‑reperfusion injury (MIRI) model in rats and the protective effect of crocin on rat myocardial tissue was observed. The levels of the inflammatory cytokines, interleukin (IL)‑1β, IL‑6 and tumor necrosis factor α (TNFα), in the sham, MI model, MI + crocin (100 mg/kg) and MI + crocin (200 mg/kg) groups were compared in the rat myocardial tissue. The TUNEL assay was used to detect apoptosis of myocardial cells. In addition, RAW264.7 cells were stimulated with the inflammatory factors recombinant mouse high mobility group box 1 (rmHMGB1) and recombinant mouse heat shock protein 60 (rmHSP60). The inhibitory effect of crocin on inflammatory cytokine levels was observed using ELISA. Western blotting was used to detect the inhibitory effect of crocin on KBTBD7. The inhibitory effect of KBTBD7 knockdown on MAPK and nuclear factor (NF)‑κB signaling pathways was also analyzed. The expression levels of IL‑1β, IL‑6 and TNFα were significantly decreased in the crocin‑treated groups compared with in the model group. Crocin significantly reduced the apoptosis of myocardial cells and significantly inhibited the release of inflammatory cytokines induced by rmHMGB1 and rmHSP60. KBTBD7 was determined to be a target of crocin. Knockdown of KBTBD7 significantly inhibited p38 and NF‑κB signaling pathways. Furthermore, the results demonstrated that KBTBD7 knockdown significantly reduced the production of inflammatory cytokines induced by rmHMGB1 and rmHSP60. KBTBD7 knockdown also significantly reduced p38 and NF‑κB signaling in the rmHMGB1‑ and rmHSP60‑treated groups. The present study demonstrated the potential protective effect of crocin on MIRI in rats. The underlying mechanism may be through direct inhibition of KBTBD7, thereby inhibiting excessive inflammatory responses and myocardial cell apoptosis following myocardial infarction.
心肌梗死（MI）是指由冠状动脉疾病引起的心肌缺血性坏死。值得注意的是，藏红花素对心脏具有保护作用。本研究旨在：i）研究藏红花素（一种番红花属植物中的活性成分）对心肌缺血的保护作用；ii）验证藏红花素与含kelch重复序列和BTB结构域蛋白7（KBTBD7）之间的相互作用，KBTBD7是BTB-kelch蛋白家族的一个新成员。在本研究中，结扎大鼠左冠状动脉前降支以建立心肌缺血-再灌注损伤（MIRI）模型，并观察藏红花素对大鼠心肌组织的保护作用。比较假手术组、MI模型组、MI + 藏红花素（100 mg/kg）组和MI + 藏红花素（200 mg/kg）组大鼠心肌组织中炎性细胞因子白细胞介素（IL）-1β、IL-6和肿瘤坏死因子α（TNFα）的水平。采用TUNEL法检测心肌细胞凋亡。此外，用炎性因子重组小鼠高迁移率族蛋白B1（rmHMGB1）和重组小鼠热休克蛋白60（rmHSP60）刺激RAW264.7细胞。用ELISA观察藏红花素对炎性细胞因子水平的抑制作用。用蛋白质印迹法检测藏红花素对KBTBD7的抑制作用。还分析了KBTBD7基因敲低对丝裂原活化蛋白激酶（MAPK）和核因子（NF）-κB信号通路的抑制作用。与模型组相比，藏红花素处理组中IL-1β、IL-6和TNFα的表达水平显著降低。藏红花素显著减少心肌细胞凋亡，并显著抑制rmHMGB1和rmHSP60诱导的炎性细胞因子释放。确定KBTBD7是藏红花素的一个靶点。敲低KBTBD7可显著抑制p38和NF-κB信号通路。此外，结果表明，敲低KBTBD7可显著减少rmHMGB1和rmHSP60诱导的炎性细胞因子产生。敲低KBTBD7还可显著降低rmHMGB1和rmHSP60处理组中的p38和NF-κB信号。本研究证明了藏红花素对大鼠MIRI具有潜在保护作用。其潜在机制可能是通过直接抑制KBTBD7，从而抑制心肌梗死后过度的炎症反应和心肌细胞凋亡。
