University of Texas at San Antonio, One UTSA Circle, San Antonio, TX 78249, USA.
Curr Eye Res. 2012 Nov;37(11):1045-53. doi: 10.3109/02713683.2012.713152. Epub 2012 Aug 20.
Acrolein has been implicated in retinal pigment epithelium (RPE) cell death, and has been associated with diabetic retinopathy. Our purpose was to investigate the potential effect of high glucose in influencing acrolein-mediated RPE cytokine production and cell death. We investigated the influence of the acrolein effect on ARPE-19 cells in high glucose conditions and quantified the release of transforming growth factor β (TGFβ1 and 2) and vascular endothelial growth factor (VEGF). We assessed the ability of N-benzylhydroxylamine(NBHA) as well as TGFβ pathway inhibitors SIS3 and SB431542 to prevent this effect of acrolein on ARPE-19 cells.
Confluent ARPE-19 cells were treated with acrolein and/or NBHA in both 5.5 and 18.8 mM glucose conditions. Cells were also pretreated with SIS3, a specific inhibitor of the SMAD3 pathway, and SB431542, a specific inhibitor of TGFβ signaling pathway, before treating them with acrolein. Viable cells were counted and ELISAs were performed to measure the cytokines TGFβ1 and 2, and VEGF released into the conditioned media.
In ARPE-19 cells exposed to acrolein and hyperglycemia there was reduced cell viability and an increase in the cell media of VEGF, TGFβ1, and TGFβ2, which was reversed by NBHA. Acrolein/hyperglycemia-induced cell viability reduction and cytokine overproduction was also reduced by TGFβ pathway blockade.
We conclude that the effect of acrolein on the reduction of viability and VEGF increase by ARPE-19 cells in hyperglycemic media is conducted through the TGFβ signaling pathway. Our results suggest that benefits of sequestering acrolein by NBHA and the blockage of the TGFβ pathway by SB431542 and SIS3 offer suggestions as to potential useful pharmacological drug candidates for the prevention of diabetes-induced complications in the eye.
丙烯醛已被牵连到视网膜色素上皮 (RPE) 细胞死亡，并与糖尿病视网膜病变有关。我们的目的是研究高葡萄糖对丙烯醛介导的 RPE 细胞因子产生和细胞死亡的潜在影响。我们研究了丙烯醛在高葡萄糖条件下对 ARPE-19 细胞的影响，并量化了转化生长因子β(TGFβ1 和 2)和血管内皮生长因子 (VEGF)的释放。我们评估了 N-苄基羟胺 (NBHA) 以及 TGFβ 途径抑制剂 SIS3 和 SB431542 阻止丙烯醛对 ARPE-19 细胞这种作用的能力。
在 5.5 和 18.8 mM 葡萄糖条件下，用丙烯醛和/或 NBHA 处理汇合的 ARPE-19 细胞。在用丙烯醛处理细胞之前，还预先用 SMAD3 途径的特异性抑制剂 SIS3 和 TGFβ 信号通路的特异性抑制剂 SB431542 预处理细胞。计数存活细胞，并通过 ELISA 测量条件培养基中释放的细胞因子 TGFβ1 和 2 和 VEGF。
在暴露于丙烯醛和高血糖的 ARPE-19 细胞中，细胞活力降低，细胞培养基中的 VEGF、TGFβ1 和 TGFβ2 增加，这被 NBHA 逆转。TGFβ 途径阻断也减少了丙烯醛/高血糖诱导的细胞活力降低和细胞因子过度产生。
我们得出结论，丙烯醛在高糖培养基中对 ARPE-19 细胞活力降低和 VEGF 增加的影响是通过 TGFβ 信号通路进行的。我们的结果表明，通过 NBHA 隔离丙烯醛和通过 SB431542 和 SIS3 阻断 TGFβ 途径提供了一些有希望的药理学药物候选物，用于预防糖尿病引起的眼部并发症。
