白藜芦醇对高氧诱导早产儿外周血单个核细胞氧化应激损伤的影响及其机制研究

Resveratrol increases sirtuin 1 expression in peripheral blood mononuclear cells of premature infants and inhibits the oxidative stress induced by hyperoxia in vivo

摘要:

目的 探讨沉默接合型信息调节因子2 同源蛋白1(SIRT1)激动剂白藜芦醇(Res)对高氧诱导早产儿外周血单个核细胞(PBMC)中SIRT1 表达和活性氧簇(ROS)产生的影响。方法 采集胎龄<32 周且未吸氧的早产儿外周血分离PBMC,随机分为对照组、空气+Res 组、高氧组和高氧+Res 组,在体外建模及培养48 h 后,采用激光共聚焦显微镜检测PBMC 内ROS 水平,全光谱分光光度仪检测培养液中丙二醛(MDA)含量,细胞免疫荧光检测SIRT1 定位,Western blot 检测PBMC 内SIRT1 蛋白表达水平。结果 与对照组相比,空气+Res 组中SIRT1 表达水平明显增加(P<0.05);高氧组中ROS、MDA 水平及SIRT1 转位率明显增加(P<0.05),SRIT1 蛋白表达水平明显降低(P<0.05)。与高氧组相比,高氧+Res 组中ROS、MDA 水平及SIRT1 转位率明显降低(P<0.05),SIRT1 蛋白表达水平明显增加(P<0.05)。结论 在高氧诱导下,Res 可以通过提高早产儿PBMC 的SIRT1 表达并抑制SIRT1 核-浆穿梭从而增加早产儿抗氧化应激的能力,进而减少早产儿氧化应激损伤。

Abstract:

Objective To explore the effect of resveratrol on the levels of sirtuin 1 (SIRT1) and reactive oxygen species (ROS) in peripheral blood mononuclear cells (PBMCs) of premature infants exposed to hyperoxia. Methods Peripheral blood and isolated PBMCs from premature infants (gestational age <32 weeks) without oxygen supplement were collected and were randomly assigned into four groups: control, air+resveratrol, hyperoxia, and hyperoxia + resveratrol. The PBMCs were cultured in vitro for 48 hours, then the ROS content in PBMCs was measured by laser scanning confocal microscopy. Malondialdehyde (MDA) content in the medium was measured by the whole spectrum spectrophotometer. SIRT1 positioning was assessed by immunofluorescence. SIRT1 expression levels in PBMCs were measured by Western bolt. Results Compared with the control group, the level of SIRT1 in the air+resveratrol group increased significantly (P<0.05). The levels of ROS and MDA and the SIRT1 transposition rate in the hyperoxia group increased significantly, while the expression level of SIRT1 decreased significantly compared with the control group (P<0.05). The levels of ROS and MDA and the SIRT1 transposition rate decreased significantly (P<0.05), and the expression level of SIRT1 increased significantly in the hyperoxia+resveratrol group (P<0.05). Conclusions Resveratrol can increase SIRT1 expression in PBMCs and inhibit SIRT1 shuttle from nucleus to cytoplasm in order to increase the ability of antioxidative stress in premature infants exposed to hyperoxia, thereby reducing the oxidative stress injury in premature infants.