Rom Oren, Volkova Nina, Nandi Sukhendu, Jelinek Raz, Aviram Michael
*The Lipid Research Laboratory, Rappaport Faculty of Medicine, Technion-Israel Institute of Technology, Haifa, Israel; and †Department of Chemistry, Ben-Gurion University, Beer Sheva, Israel.
J Cardiovasc Pharmacol. 2016 Aug;68(2):106-14. doi: 10.1097/FJC.0000000000000391.
At high concentrations, polyphenols induce cell death, and the polyphenols-rich pomegranate juice (PJ), known for its antioxidative/antiatherogenic properties, can possibly affect cell death, including macrophage death involved in atherogenesis. In the present study, apoptotic/necrotic macrophage death was analyzed in J774A.1 macrophages and in peritoneal macrophages isolated from atherosclerotic apoE-/- mice treated with PJ. The effects of PJ were compared with those of the free radical generator 2, 2'-azobis (2-amidinopropane) dihydrochloride (AAPH). Both PJ and AAPH significantly increased J774A.1 macrophage death; however, flow cytometric and microscopic analyses using annexin V/propidium iodide revealed that PJ increased the early apoptosis of the macrophage dose dependently (up to 2.5-fold, P < 0.01), whereas AAPH caused dose-dependent increases in late apoptosis/necrosis (up to 12-fold, P < 0.001). Unlike PJ, AAPH-induced macrophage death was associated with increased intracellular oxidative stress (up to 7-fold, P < 0.001) and with lipid stress demonstrated by triglyceride accumulation (up to 3-fold, P < 0.01) and greater chromatic vesicle response to culture medium (up to 5-fold, P < 0.001). Accordingly, recombinant paraoxonase 1, which hydrolyzes oxidized lipids, attenuated macrophage death induced by AAPH, but not by PJ. Similar apoptotic and oxidative effects were found in macrophages from apoE-/- mice treated with PJ or AAPH. As macrophage apoptotic/necrotic death has considerable impact on atherosclerosis progression, these findings may provide novel mechanisms for the antiatherogenicity of PJ.
在高浓度时，多酚会诱导细胞死亡，而富含多酚的石榴汁（PJ）以其抗氧化/抗动脉粥样硬化特性而闻名，它可能会影响细胞死亡，包括参与动脉粥样硬化形成的巨噬细胞死亡。在本研究中，分析了J774A.1巨噬细胞以及从经PJ处理的动脉粥样硬化载脂蛋白E基因敲除（apoE-/-）小鼠分离的腹腔巨噬细胞中的凋亡/坏死性巨噬细胞死亡情况。将PJ的作用与自由基发生器2,2'-偶氮二（2-脒基丙烷）二盐酸盐（AAPH）的作用进行了比较。PJ和AAPH均显著增加J774A.1巨噬细胞死亡；然而，使用膜联蛋白V/碘化丙啶进行的流式细胞术和显微镜分析显示，PJ剂量依赖性地增加巨噬细胞的早期凋亡（高达2.5倍，P<0.01），而AAPH导致晚期凋亡/坏死剂量依赖性增加（高达12倍，P<0.001）。与PJ不同，AAPH诱导的巨噬细胞死亡与细胞内氧化应激增加（高达7倍，P<0.001）以及甘油三酯积累所显示的脂质应激（高达3倍，P<0.01）和对培养基更大的色泡反应（高达5倍，P<0.001）相关。因此，水解氧化脂质的重组对氧磷酶1减弱了AAPH诱导的巨噬细胞死亡，但未减弱PJ诱导的巨噬细胞死亡。在用PJ或AAPH处理的apoE-/-小鼠的巨噬细胞中也发现了类似的凋亡和氧化作用。由于巨噬细胞凋亡/坏死性死亡对动脉粥样硬化进展有相当大的影响，这些发现可能为PJ的抗动脉粥样硬化作用提供新机制。
