Abstract：
Adalimumab (ADA) is an anti-inflammatory antibody that has FDA approval as a systemic medication for treating noninfectious uveitis. It is also provisionally being investigated as an intravitreal injection for various retinal conditions. This study aimed to assess the effect of ADA on apoptotic, inflammatory, and fibrogenesis gene expression at mRNA and protein levels in retinal pigment epithelial (RPE) cells. RPEs were treated with serial concentrations of ADA (0.5x, x, 2x, and 4x; [x = 250 µg/mL]) for 24 hours. MTT assay was done and the mRNA and protein expressions were quantified using real-time PCR and ELISA assay, respectively. The mRNA levels of IL-1b and IL-6 were significantly increased in ADA-treated RPEs at 0.5x and x concentrations. However, the increase in cytokine secretion was observed only in IL-1b at x concentration. TGF-β was significantly upregulated in the 0.5x and 4x doses of ADA both at mRNA and protein levels. MTT assay, along with an unchanged BCL-2/BAX ratio confirmed the safety of ADA on RPEs at all studied concentrations. In conclusion, despite its safety, the 2x concentration of ADA was the only dose that did not ignite the expression of any of the studied inflammatory and fibrogenesis genes. This dosage, which is roughly equal to 2 mg intravitreal dose in a clinical setting, might be referred to as a reference starting point for future in-vivo studies in ocular conditions.
摘要：
阿达木单抗（ADA）是一种抗炎抗体，已被FDA批准作为治疗非感染性葡萄膜炎的全身性药物。它也被暂时作为用于各种视网膜病症的玻璃体内注射进行研究。本研究旨在评估ADA对细胞凋亡的影响，炎症，视网膜色素上皮（RPE）细胞中mRNA和蛋白质水平的纤维发生基因表达。用连续浓度的ADA处理RPE（0.5x，x,2x,和4x；[x=250µg/mL]）持续24小时。MTT法检测mRNA和蛋白的表达,采用实时PCR和ELISA法,分别。在ADA处理的RPE中,IL-1b和IL-6的mRNA水平在0.5x和x浓度下显著增加。然而,在x浓度下,仅在IL-1b中观察到细胞因子分泌的增加。在mRNA和蛋白质水平上,TGF-β在ADA的0.5x和4x剂量中显著上调。MTT测定，BCL-2/BAX比值不变,证实了ADA在所有研究浓度下对RPE的安全性.总之,尽管安全,2x浓度的ADA是唯一不引起任何研究的炎症和纤维发生基因表达的剂量。这个剂量，在临床上大约等于2毫克玻璃体内剂量，可能被称为未来眼部疾病体内研究的参考起点。
