Liu Yuefeng, Luo Weimin, Luo Xiangyu, Yong Zhang, Zhong Xiaodong
Department of Ophthalmology, Taihe Hospital of Shiyan Affiliated to Hubei University of Medicine Shiyan 442000, Hubei, China.
Department of Cardiothoracic Surgery, Taihe Hospital of Shiyan Affiliated to Hubei University of Medicine Shiyan 442000, Hubei, China.
Int J Clin Exp Med. 2015 Sep 15;8(9):15759-65. eCollection 2015.
This study was to investigate the effects of Rosa laevigata Michx. (RLM) extract on reactive oxygen species (ROS) production and mitochondrial membrane potential (MMP) in lens epithelial cells (LECs) cultured under high glucose. SRA01/04 cell models of diabetic cataract were established by high glucose culture, and these cells were administrated with RLM extract. Levels of ROS and MMP in SRA01/04 cells were detected with fluorescent probes. HO-1 expression level, Akt phosphorylation, and Nrf2 translocation were detected by Western blot. RLM treatment could exert protective effects on SRA01/04 cells under high glucose condition, decreasing the ROS production and elevating the MMP in these cells. Western blot analysis indicated that the expression level of HO-1 was significantly elevated in SRA01/04 cells after the RLM treatment. Moreover, when HO-1 was interfered with siRNA, the effects of RLM on the levels of ROS and MMP were diminished, indicating that HO-1 induction was necessary for the function of RLM. Furthermore, HO-1 inducing effects of RLM were mediated by the PI3K/Akt and Nrf2/ARE pathways. The effects would be abolished when either pathway was inhibited by pharmacological manipulation or siRNA silencing. RLM inhibits ROS production and elevates MMP, through the induction of HO-1 expression, in LECs under high glucose condition. The protective effects of RLM are mediated by the PI3K/Akt and Nrf2/ARE pathways. Our findings provide theoretic basis and experimental evidence for the application of RLM in the treatment of diabetic cataract.
本研究旨在探讨金樱子提取物对高糖培养的晶状体上皮细胞(LECs)中活性氧(ROS)生成及线粒体膜电位(MMP)的影响。通过高糖培养建立糖尿病性白内障的SRA01/04细胞模型,并对这些细胞给予金樱子提取物处理。用荧光探针检测SRA01/04细胞中ROS和MMP的水平。通过蛋白质免疫印迹法检测血红素加氧酶-1(HO-1)的表达水平、蛋白激酶B(Akt)磷酸化及核因子E2相关因子2(Nrf2)的转位。金樱子处理可对高糖条件下的SRA01/04细胞发挥保护作用,减少这些细胞中ROS的生成并提高MMP。蛋白质免疫印迹分析表明,金樱子处理后SRA01/04细胞中HO-1的表达水平显著升高。此外,当用小干扰RNA(siRNA)干扰HO-1时,金樱子对ROS和MMP水平的影响减弱,表明HO-1的诱导对于金樱子的功能是必需的。此外,金樱子诱导HO-1的作用是由磷脂酰肌醇-3激酶(PI3K)/Akt和Nrf2/抗氧化反应元件(ARE)通路介导的。当通过药理学操作或siRNA沉默抑制任一通路时,这些作用均会被消除。在高糖条件下,金樱子通过诱导HO-1的表达来抑制LECs中ROS的生成并提高MMP。金樱子的保护作用由PI3K/Akt和Nrf2/ARE通路介导。我们的研究结果为金樱子在糖尿病性白内障治疗中的应用提供了理论依据和实验证据。