Department of Ophthalmology, Duke Eye Center, Duke University Medical Center, Durham, North Carolina 27710, USA.
Invest Ophthalmol Vis Sci. 2011 Jun 21;52(7):4395-401. doi: 10.1167/iovs.10-6652.
To evaluate the potential role that FoxO transcription factors play in modulating resveratrol's protective effects against oxidative stress in lens epithelial cells.
Primary human or porcine lens epithelial cells (LECs) were treated with resveratrol (RES) 25 μM and incubated under either physiologic (5%) or chronic hyperoxic (40%) oxygen conditions. Acute oxidative stress was applied using 600 μM H(2)O(2). Changes in expression of FoxO1A, FoxO3A, and FoxO4 were analyzed. The production of intracellular reactive oxygen species (iROS), SA-β-galactosidase (SA-β-gal) activity, and autofluorescence (AF) was assessed by flow cytometry. SiRNAs of FoxO1A, FoxO3A, and FoxO4 were used to study the roles that these transcription factors play in resveratrol's protective effects against cell death induced by oxidative stress.
RES incubation under 40% oxygen increased the expression of FoxO1A, FoxO3A, and FoxO4. RES also increases mitochondrial membrane potential under 5% and/or 40% O(2) conditions and significantly decreased iROS, SA-β-gal, and AF normally induced by hyperoxic conditions. While RES had a mild pro-apoptotic effect in nonstressed cells, it significantly prevented apoptosis induced by H(2)O(2) stress. SiRNA inhibition of FoxO1A, FoxO3A, and FoxO4 not only led to loss of the anti-apoptotic effects of RES in stressed cells but actually exhibited a mild pro-apoptotic effect.
RES exerts a protective effect against oxidative damage in LEC cultures. The levels of expression of FoxO1A, FoxO3A, and FoxO4 appear to play a central role in determining the pro- or anti-apoptotic effects of RES. This has implications for future studies on oxidative stress-related lenticular disorders such as cataract formation.
评估 FoxO 转录因子在调节白藜芦醇对晶状体上皮细胞氧化应激的保护作用中的潜在作用。
用白藜芦醇（RES）25μM 处理原代人或猪晶状体上皮细胞（LEC），并在生理（5%）或慢性高氧（40%）条件下孵育。用 600μM H2O2 施加急性氧化应激。分析 FoxO1A、FoxO3A 和 FoxO4 的表达变化。通过流式细胞术评估细胞内活性氧（iROS）、SA-β-半乳糖苷酶（SA-β-gal）活性和自发荧光（AF）的产生。使用 FoxO1A、FoxO3A 和 FoxO4 的 siRNA 研究这些转录因子在白藜芦醇对氧化应激诱导的细胞死亡的保护作用中的作用。
在 40%氧气下孵育 RES 会增加 FoxO1A、FoxO3A 和 FoxO4 的表达。RES 还会增加 5%和/或 40%O2 条件下的线粒体膜电位，并显著降低高氧条件下通常诱导的 iROS、SA-β-gal 和 AF。虽然 RES 在非应激细胞中有轻微的促凋亡作用，但它能显著预防 H2O2 应激诱导的凋亡。FoxO1A、FoxO3A 和 FoxO4 的 siRNA 抑制不仅导致应激细胞中 RES 的抗凋亡作用丧失，而且实际上表现出轻微的促凋亡作用。
RES 对 LEC 培养物中的氧化损伤具有保护作用。FoxO1A、FoxO3A 和 FoxO4 的表达水平似乎在决定 RES 的促凋亡或抗凋亡作用中起核心作用。这对未来与氧化应激相关的晶状体疾病（如白内障形成）的研究具有重要意义。
