[关键词]
[摘要]
目的 基于动物心肌缺血模型及体外心肌细胞损伤模型探究复方丹参滴丸调控脂氧合酶(lipoxygenase,LOX)-核因子-κB(nuclear factor-κB,NF-κB)途径抑制炎症反应治疗心肌缺血的作用机制。方法 采用冠状动脉左前降支结扎术制备大鼠心肌缺血模型,心电图检测心肌缺血情况;Western blotting检测心肌组织LOX途径关键酶LOX5、LOX12、LOX15及下游分子p-NF-κB p65和白细胞介素-1β(interleukin-1β,IL-1β)的蛋白表达。体外培养H9c2心肌细胞,采用缺氧缺糖6 h诱导心肌细胞损伤模型,荧光倒置显微镜下观察心肌细胞形态;MTT法检测细胞活力;Western blotting检测LOX-NF-κB途径相关蛋白表达。结果 复方丹参滴丸显著抑制心肌缺血大鼠心肌组织中LOX5、LOX12、LOX15和IL-1β的蛋白表达及NF-κB p65的磷酸化水平(P<0.01、0.001)。体外细胞实验结果显示复方丹参滴丸明显改善H9c2心肌细胞缺氧缺糖损伤,显著提高心肌细胞存活率(P<0.01、0.001),降低LOX5、LOX12、LOX15和IL-1β的蛋白表达及NF-κB p65的磷酸化水平(P<0.05、0.01、0.001)。结论 复方丹参滴丸可通过抑制LOX-NF-κB炎症途径减轻心肌缺血损伤。
[Key word]
[Abstract]
Objective To explore the mechanism of Compound Danshen Dripping Pills (复方丹参滴丸) on treatment of myocardial ischemia by regulating lipoxygenase (LOX)-nuclear factor-κB (NF-κB) pathway to inhibit inflammatory response based on animal models of myocardial ischemia and in vitro myocardial cell injury model. Methods The rat model of myocardial ischemia was established by ligation of left anterior descending coronary artery, with myocardial ischemia detected by ECG; Protein expressions of LOX5, LOX12, LOX15, p-NF-κB p65 and interleukin-1β (IL-1β) were detected by Western blotting. H9c2 cardiomyocytes were cultured in vitro, and the model of myocardial cell injury was induced by deprivation of oxygen-glucose (OGD) for 6 h, morphology of myocardial cells was observed under fluorescence inverted microscope; Cell viability was detected by MTT; Western blotting was used to detect LOX-NF-κB pathway related protein expressions.Results Compound Danshen Dripping Pills significantly inhibited the protein expression levels of LOX5, LOX12, LOX15, p-NF-κB p65 and IL-1β in myocardial tissues of rats with ischemia (P < 0.01, 0.001). The in vitro cell experiments indicated that Compound Danshen Dripping Pills could dramatically improve the OGD injury of H9c2 myocardial cells, significantly increase the survival rate of myocardial cells (P < 0.01, 0.001), and reduce LOX5, LOX12, LOX15 and IL-1β protein expressions and NF-κB p65 phosphorylation level (P < 0.05, 0.01, 0.001). Conclusion Compound Danshen Dripping Pills can alleviate myocardial ischemia injury by inhibiting LOX-NF-κB inflammatory pathway.
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[基金项目]
国家自然科学基金资助项目(82004095);国家自然科学基金资助项目(81903950);国家自然科学基金资助项目(81904169);中央高校基本科研业务费专项(2021-JYB-XJSJJ031,2019-JYB-JS-095,2022-JYB-XJSJJ008)