Soltani Behrooz, Bodaghabadi Narges, Ghaemi Nasser, Sadeghizadeh Majid
a Department of Biotechnology , College of Science, University of Tehran , Tehran , Iran.
b Department of Genetics, Faculty of Biological Sciences , Tarbiat Modares University , Tehran , Iran.
Int J Radiat Biol. 2017 Mar;93(3):303-314. doi: 10.1080/09553002.2016.1242817. Epub 2016 Nov 3.
We examined the potential of a dendrosomal nanoformulation of curcumin (DNC) for intervention of ionizing radiation (IR)-induced damage (particularly leading to atherosclerosis), employing an irradiated THP-1 macrophage model.
Differentiated THP-1 macrophages were irradiated and treated with curcumin or DNC nanoformulation (and oxidized low density lipoprotein, ox-LDL, to promote foam cells). Chemical, biochemical, and genetics tools including viability and apoptosis, multiple ELISA, real-time PCR, Western blotting, enzyme activity, and fluorimetry assays were employed to illustrate IR damage as well as the DNC intervention potential.
DNC per se at 10 μM exerted no cytotoxic effects on macrophages. However, it caused apoptosis in 2 Gy-irradiated macrophages which were treated with ox-LDL, chiefly through a caspase-dependent pathway involving caspase-3. Concurrently, 10 μM DNC prevented the IR-induced rise in lipid accumulation (72% decrease compared to IR control, p < .0001), dil-oxLDL uptake (78% decrease, p < .005), protein and mRNA expression of cholesterol influx genes, CD36 and SR-A, NF-κB activation (81% less binding activity, p < .001; and lower nuclear presence of p65), cytokine (monocyte chemoattractant protein-1 and interleukin-1β) release, reactive oxygen species (ROS), and oxidative damage to DNA (37% decrease in 8-OHdG, p < .05) and lipids (62% decrease in 8-isoprostane, p < .005). DNC facilitated the uptake of curcumin in irradiated macrophages, increased glutathione peroxidase expression and activity, restored glutathione (GSH) level, and upregulated the expression of a cholesterol efflux gene, ABCA1. Two other antioxidants, resveratrol and N-acetyl cycteine (NAC), could simulate some of the beneficial effects of DNC against IR-induced CD36 expression and lipid accumulation, which were obviated by buthionine sulfoximine (BSO) pre-treatment of macrophages. However, some modulatory effects of DNC, particularly on lipid accumulation and the expression of SR-A and ABCA1 genes, seemed to be independent of its antioxidant effect, since they were still observed in BSO-pretreated macrophages, depleted of GSH.
DNC treatment suppresses IR-induced oxidative damage, inflammation, and foam cell formation in macrophages through multiple mechanisms.
我们使用经辐照的THP-1巨噬细胞模型，研究了姜黄素的树枝状纳米制剂（DNC）干预电离辐射（IR）诱导损伤（特别是导致动脉粥样硬化的损伤）的潜力。
将分化的THP-1巨噬细胞进行辐照，并用姜黄素或DNC纳米制剂处理（以及氧化低密度脂蛋白，即ox-LDL，以促进泡沫细胞形成）。采用包括细胞活力和凋亡、多种酶联免疫吸附测定（ELISA）、实时聚合酶链反应（PCR）、蛋白质印迹法、酶活性测定和荧光测定等化学、生化及遗传学工具，以阐明IR损伤以及DNC的干预潜力。
10 μM的DNC本身对巨噬细胞无细胞毒性作用。然而，它导致经2 Gy辐照且用ox-LDL处理的巨噬细胞发生凋亡，主要通过涉及半胱天冬酶-3的半胱天冬酶依赖性途径。同时，10 μM的DNC可防止IR诱导的脂质积累增加（与IR对照组相比降低72%，p <.0001）、二碘氧化低密度脂蛋白（dil-oxLDL）摄取增加（降低78%，p <.005）、胆固醇内流基因CD36和清道夫受体A（SR-A）的蛋白质和mRNA表达增加、核因子κB（NF-κB）激活（结合活性降低81%，p <.001；且p65的核内存在减少）、细胞因子（单核细胞趋化蛋白-1和白细胞介素-1β）释放、活性氧（ROS）以及对DNA的氧化损伤（8-羟基脱氧鸟苷降低37%，p <.05）和脂质氧化损伤（8-异前列腺素降低62%，p <.005）。DNC促进了辐照巨噬细胞对姜黄素的摄取，增加了谷胱甘肽过氧化物酶的表达和活性，恢复了谷胱甘肽（GSH）水平，并上调了胆固醇外流基因ABCA1的表达。另外两种抗氧化剂白藜芦醇和N-乙酰半胱氨酸（NAC）可模拟DNC对IR诱导的CD36表达和脂质积累的一些有益作用，而巨噬细胞经丁硫氨酸亚砜胺（BSO）预处理可消除这些作用。然而，DNC的一些调节作用，特别是对脂质积累以及SR-A和ABCA1基因表达的调节作用，似乎与其抗氧化作用无关，因为在经BSO预处理且GSH耗竭的巨噬细胞中仍可观察到这些作用。
DNC处理可通过多种机制抑制IR诱导的巨噬细胞氧化损伤、炎症和泡沫细胞形成。
