State Key Laboratory of Functions and Applications of Medicinal Plants, Engineering Research Center for the Development and Application of Ethnic Medicine and Traditional Chinese Medicine (Ministry of Education), Guizhou Medical University, Guiyang 550004, China.
School of Pharmacy, Guizhou Medical University, Guiyang 550004, China.
J Tradit Chin Med. 2021 Feb;41(1):51-58. doi: 10.19852/j.cnki.jtcm.2021.01.007.
To investigate the efficacy of water fraction from Dioscorea cirrhosa (WF) on oxidative damage and apoptosis of cardiomyocytes induced by H2O2, and to study its mechanism.
Cell viability was measured by the MST assay kit. The content of malondialdehyde (MDA), release of lactate dehydrogenase (LDH) and activity of catalase (CAT) and superoxide dismutase (SOD) were detected by biochemical kit. The content of reactive oxygen species (ROS) was assessed by nonfluorescent probe 2' ,7'-dichlorofluorescin diacetate (DCFH-DA). JC-1 was used to analyze the mitochondrial membrane potential (mtΔΨ) and Annexin-V-FITC/PI staining was applied to assess apoptosis of H9c2 by flow cytometry. Moreover, the expression of B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X (Bax), caspase-3, caspase-9, cleaved-caspase-3 and cleaved-caspase -9 proteins was determined by western blot analysis.
WF increased cell viability and decreased LDH leakage in H9c2 cells exposed to H2O2. WF treatment decreased ROS and MDA level, enhanced SOD and CAT activities, improved mtΔΨ and inhibited apoptosis. Western blot analysis demonstrated that the ratio of Bcl-2/Bax was increased and the expression cleaved-caspase-3, caspase-3, cleaved-caspase-9 and caspase-9 were decreased in group treated with WF.
WF protects H9c2 myocardial cells on H2O2-induced oxidative stress and apoptosis by scavenging ROS, improving antioxidant capacity, protecting mitochondrial and regulating the proteins expression related to apoptosis.
研究黄姜水提物（WF）对 H2O2 诱导的心肌细胞氧化损伤和凋亡的作用，并探讨其机制。
采用 MST 试剂盒检测细胞活力。采用生化试剂盒检测丙二醛（MDA）含量、乳酸脱氢酶（LDH）释放、过氧化氢酶（CAT）和超氧化物歧化酶（SOD）活性。采用非荧光探针 2',7'-二氯荧光素二乙酸酯（DCFH-DA）评估活性氧（ROS）含量。JC-1 用于分析线粒体膜电位（mtΔΨ），流式细胞术采用 Annexin-V-FITC/PI 染色评估 H9c2 细胞的凋亡。此外，采用 Western blot 分析检测 B 细胞淋巴瘤-2（Bcl-2）、Bcl-2 相关 X（Bax）、半胱氨酸天冬氨酸蛋白酶-3（caspase-3）、半胱氨酸天冬氨酸蛋白酶-9（caspase-9）、cleaved-caspase-3 和 cleaved-caspase-9 蛋白的表达。
WF 增加了 H2O2 处理的 H9c2 细胞的活力并降低了 LDH 漏出。WF 处理降低了 ROS 和 MDA 水平，增强了 SOD 和 CAT 活性，改善了 mtΔΨ 并抑制了凋亡。Western blot 分析表明，WF 处理组的 Bcl-2/Bax 比值增加，cleaved-caspase-3、caspase-3、cleaved-caspase-9 和 caspase-9 的表达减少。
WF 通过清除 ROS、提高抗氧化能力、保护线粒体和调节与凋亡相关的蛋白表达，保护 H9c2 心肌细胞免受 H2O2 诱导的氧化应激和凋亡。
