Poostchi Ophthalmology Research Center, Department of Ophthalmology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Islamic Republic of Iran.
Biomed Pharmacother. 2024 May;174:116568. doi: 10.1016/j.biopha.2024.116568. Epub 2024 Apr 9.
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.
阿达木单抗（ADA）是一种抗炎抗体，已获得美国食品药品监督管理局（FDA）批准，作为治疗非感染性葡萄膜炎的全身性药物。它也被临时用作各种视网膜疾病的玻璃体内注射。本研究旨在评估 ADA 在视网膜色素上皮（RPE）细胞的 mRNA 和蛋白质水平上对细胞凋亡、炎症和纤维化基因表达的影响。用不同浓度的 ADA（0.5x、x、2x 和 4x；[x=250μg/mL]）处理 RPE 24 小时。进行 MTT 检测，并分别使用实时 PCR 和 ELISA 检测来定量 mRNA 和蛋白质表达。在 0.5x 和 x 浓度的 ADA 处理的 RPE 中，IL-1b 和 IL-6 的 mRNA 水平显著增加。然而，仅在 x 浓度下观察到细胞因子分泌增加。TGF-β在 ADA 的 0.5x 和 4x 剂量下均在 mRNA 和蛋白质水平上显著上调。MTT 检测以及 BCL-2/BAX 比值不变证实 ADA 在所有研究浓度下对 RPE 均安全。总之，尽管 ADA 安全，但只有 2x 浓度的 ADA 不会引发任何研究的炎症和纤维化基因的表达。该剂量大约相当于临床中 2mg 的玻璃体内剂量，可能是未来眼部疾病体内研究的参考起点。
