Jiangxi Provincial Institute of Hypertension, First Affiliated Hospital of Nanchang University, Nanchang 330006, China.
Jiangxi Provincial Key Laboratory of Basic Pharmacology, Nanchang University School of Pharmaceutical Science, Nanchang 330006, China.
Biomed Pharmacother. 2020 Jun;126:110040. doi: 10.1016/j.biopha.2020.110040. Epub 2020 Mar 4.
Kaempferol (Kae), a flavonoid, has been found in fruits and other vegetables, possesses many biological activities. 14-3-3 protein exerts protection on various types of injured tissues and cells. Doxorubicin (Dox) causes excessive reactive oxygen species (ROS) generation, which induces endotheliotoxicity and cardiotoxicity. We hypothesized that Kae could protect vascular endothelium by regulating 14-3-3γ or related pathways against Dox toxicity. HUVECs were established Dox-toxic injury models. Kae's effects were assessed with many physiological, enzymatic, cellular, and molecular biological indexes. Our results showed that Dox-induced damage in HUVECs were reduced through Kae to promote the expression of total protein 14-3-3γ and mitochondrial Bcl-2, phosphorylate Bad, increase cell viability, NO content, DDAHⅡactivity, p-eNOS/eNOS ratio, and MMP levels, maintained NAD/NADH and GSH/GSSG balance, and decrease LDH and caspase-3 activities, ADMA content, ROS generation, mPTP openness, and apoptosis. Kae's effects were abolished with pAD/14-3-3γ-shRNA downregulating 14-3-3γ expression, or ABT-737 inhibiting Bcl-2 activity. This study demonstrated that Kae protected the vascular endothelium against Dox-induced damage by regulating 14-3-3γ and ADMA/DDAHⅡ/eNOS/NO pathway, inhibiting oxidative stress, and improving mitochondrial function.
山奈酚（Kae）是一种存在于水果和其他蔬菜中的类黄酮，具有多种生物活性。14-3-3 蛋白对各种受损组织和细胞具有保护作用。多柔比星（Dox）会导致过量的活性氧（ROS）生成，从而引起内皮毒性和心脏毒性。我们假设 Kae 可以通过调节 14-3-3γ 或相关途径来抵抗 Dox 毒性，从而保护血管内皮。建立了 HUVECs 的 Dox 毒性损伤模型。使用多种生理、酶学、细胞和分子生物学指标评估 Kae 的作用。结果表明，Kae 通过促进总蛋白 14-3-3γ 和线粒体 Bcl-2 的表达、磷酸化 Bad、增加细胞活力、NO 含量、DDAHⅡ活性、p-eNOS/eNOS 比值和 MMP 水平，维持 NAD/NADH 和 GSH/GSSG 平衡，降低 LDH 和 caspase-3 活性、ADMA 含量、ROS 生成、mPTP 开放和细胞凋亡，减轻 Dox 诱导的 HUVECs 损伤。下调 14-3-3γ 表达的 pAD/14-3-3γ-shRNA 或抑制 Bcl-2 活性的 ABT-737 可消除 Kae 的作用。本研究表明，Kae 通过调节 14-3-3γ 和 ADMA/DDAHⅡ/eNOS/NO 途径、抑制氧化应激和改善线粒体功能来保护血管内皮免受 Dox 诱导的损伤。
