Jiang Qiao, Pan Defeng, Yang Yu, Hu Ya, Fang Liang, Shang Pingping, Xia Yong, Li Dongye
Institute of Cardiovascular Disease Research, Xuzhou Medical University, Xuzhou, Jiangsu, China.
Department of Cardiology, the Affiliated Hospital of Xuzhou Medical University, Xuzhou, Jiangsu, China.
Curr Pharm Biotechnol. 2018;19(5):428-437. doi: 10.2174/1389201019666180629143251.
The aim of this study was to investigate anti-apoptotic effects of luteolin on angiotensin II-stimulated murine peritoneal macrophages and to explore its mechanisms.
The viability and cytotoxicity of murine peritoneal macrophages were assessed using the Cell Counting Kit-8 assay and measuring lactate dehydrogenase levels, respectively. Apoptotic rates were determined using Annexin V/propidium iodide staining. Protein expression was examined by western blotting, and markers of macrophage phenotypes were analyzed by flow cytometry and ELISA. Luteolin decreased the apoptotic rate of angiotensin II-stimulated macrophages. This effect was associated with increased Bcl-2 and caspase-3 levels as well as decreased Bax and cleaved caspase-3 levels. Additionally, luteolin reduced the expression of M1 macrophage phenotype markers (IL-6, TNF-α, iNOS, CD16/32) and increased the expression of M2 macrophage phenotype markers (Dectin-1, IL-10, Arg-1, CD206). Moreover, luteolin blocked Akt phosphorylation on residues 308 and 473, which were up-regulated in presence of angiotensin II. The effects of luteolin were similar to those of LY294002, a specific PI3K/Akt pathway inhibitor.
These results indicated that luteolin has anti-apoptotic effects on angiotensin II-stimulated macrophages via macrophage polarization, which might be associated with PI3K/Akt signaling.
本研究旨在探讨木犀草素对血管紧张素II刺激的小鼠腹腔巨噬细胞的抗凋亡作用，并探究其机制。
分别使用细胞计数试剂盒-8检测法和测量乳酸脱氢酶水平来评估小鼠腹腔巨噬细胞的活力和细胞毒性。采用膜联蛋白V/碘化丙啶染色法测定凋亡率。通过蛋白质印迹法检测蛋白质表达，并通过流式细胞术和酶联免疫吸附测定法分析巨噬细胞表型标志物。木犀草素降低了血管紧张素II刺激的巨噬细胞的凋亡率。这种作用与Bcl-2和半胱天冬酶-3水平升高以及Bax和裂解的半胱天冬酶-3水平降低有关。此外，木犀草素降低了M1巨噬细胞表型标志物（IL-6、TNF-α、诱导型一氧化氮合酶、CD16/32）的表达，并增加了M2巨噬细胞表型标志物（树突状细胞特异性C型凝集素-1、IL-10、精氨酸酶-1、CD206）的表达。此外，木犀草素阻断了308位和473位残基上的Akt磷酸化，在血管紧张素II存在的情况下，这两个位点的磷酸化上调。木犀草素的作用与特异性PI3K/Akt通路抑制剂LY294002的作用相似。
这些结果表明，木犀草素通过巨噬细胞极化对血管紧张素II刺激的巨噬细胞具有抗凋亡作用，这可能与PI3K/Akt信号传导有关。
