Department of Cardiology, Dongzhimen Hospital Affiliated to Beijing University of Chinese Medicine, 100700, Beijing, China.
Department of Cardiology, Guanganmen Hospital, China Academy of Chinese Medical Sciences, 100053, Beijing, China.
Cell Biochem Biophys. 2024 Jun;82(2):1179-1191. doi: 10.1007/s12013-024-01267-z. Epub 2024 May 7.
Panax quinquefolius saponins (PQS) and Panax notoginseng saponins (PNS) are key bioactive compounds in Panax quinquefolius L. and Panax notoginseng, commonly used in the treatment of clinical ischemic heart disease. However, their potential in mitigating myocardial ischemia-reperfusion injury remains uncertain. This study aims to evaluate the protective effects of combined PQS and PNS administration in myocardial hypoxia/reoxygenation (H/R) injury and explore the underlying mechanisms.
To investigate the involvement of HIF-1α/BNIP3 mitophagy pathway in the myocardial protection conferred by PNS and PQS, we employed small interfering BNIP3 (siBNIP3) to silence key proteins of the pathway. H9C2 cells were categorized into four groups: control, H/R, H/R + PQS + PNS, and H/R + PQS + PNS+siBNIP3. Cell viability was assessed by Cell Counting Kit-8, apoptosis rates determined via flow cytometry, mitochondrial membrane potential assessed with the JC-1 fluorescent probes, intracellular reactive oxygen species detected with 2',7'-dichlorodihydrofluorescein diacetate, mitochondrial superoxide production quantified with MitoSOX Red, and autophagic flux monitored with mRFP-GFP-LC3 adenoviral vectors. Autophagosomes and their ultrastructure were visualized through transmission electron microscopy. Moreover, mRNA and protein levels were analyzed via real-time PCR and Western blotting.
PQS + PNS administration significantly increased cell viability, reduced apoptosis, lowered reactive oxygen species levels and mitochondrial superoxide production, mitigated mitochondrial dysfunction, and induced autophagic flux. Notably, siBNIP3 intervention did not counteract the cardioprotective effect of PQS + PNS. The PQS + PNS group showed downregulated mRNA expression of HIF-1α and BNIP3, along with reduced HIF-1α protein expression compared to the H/R group.
PQS + PNS protects against myocardial H/R injury, potentially by downregulating mitophagy through the HIF-1α/BNIP3 pathway.
西洋参总皂苷（PQS）和三七总皂苷（PNS）是西洋参和三七中关键的生物活性化合物，常用于治疗临床缺血性心脏病。然而，它们在减轻心肌缺血再灌注损伤方面的潜力尚不确定。本研究旨在评估联合使用 PQS 和 PNS 给药对心肌缺氧/复氧（H/R）损伤的保护作用，并探讨其潜在机制。
为了研究 HIF-1α/BNIP3 线粒体自噬途径在 PNS 和 PQS 赋予心肌保护中的作用，我们使用小干扰 BNIP3（siBNIP3）沉默该途径的关键蛋白。将 H9C2 细胞分为四组：对照组、H/R 组、H/R+PQS+PNS 组和 H/R+PQS+PNS+siBNIP3 组。通过 Cell Counting Kit-8 评估细胞活力，通过流式细胞术测定细胞凋亡率，通过 JC-1 荧光探针评估线粒体膜电位，通过 2',7'-二氯二氢荧光素二乙酸酯检测细胞内活性氧，通过 MitoSOX Red 定量线粒体超氧化物产生，通过 mRFP-GFP-LC3 腺病毒载体监测自噬流。通过透射电子显微镜观察自噬体及其超微结构。此外，通过实时 PCR 和 Western blot 分析 mRNA 和蛋白水平。
PQS+PNS 给药显著增加了细胞活力，减少了细胞凋亡，降低了活性氧水平和线粒体超氧化物产生，减轻了线粒体功能障碍，并诱导了自噬流。值得注意的是，siBNIP3 干预并没有抵消 PQS+PNS 的心脏保护作用。与 H/R 组相比，PQS+PNS 组 HIF-1α 和 BNIP3 的 mRNA 表达下调，HIF-1α 蛋白表达降低。
PQS+PNS 可预防心肌 H/R 损伤，可能通过下调 HIF-1α/BNIP3 通路来抑制线粒体自噬。
