Deliktaş Mehmet, Ergin Hacer, Demiray Aydın, Akça Hakan, Özdemir Özmert M A, Özdemir Mehmet Bülent
a Department of Pediatrics, Division of Neonatology, Faculty of Medicine , Pamukkale University , Denizli , Turkey.
b Department of Medical Biology, Division of Neonatology, Faculty of Medicine , Pamukkale University , Denizli , Turkey.
J Matern Fetal Neonatal Med. 2019 Jun;32(11):1813-1819. doi: 10.1080/14767058.2017.1419175. Epub 2018 Jan 2.
Unconjugated bilirubin (UCB) may cause neurotoxicity in preterm neonates due to immaturity of UGT1A1 leading to bilirubin accumulation in the brain. Caffeine used in the treatment of apnea of prematurity was reported to decrease mechanical ventilation requirement, the frequencies of bronchopulmonary dysplasia, patent ductus arteriosus, cerebral palsy and neurodevelopmental disorders in very low birth weight infants. However, the effect of caffeine on hyperbilirubinemia was not yet clarified.
We used astrocyte cell cultures obtained from 2-day-old Wistar albino rats via modified Cole and de Vellis method. UCB concentration toxic to 50% of astrocytes, and caffeine concentration increasing cell viability 100% were used in experiments. While no medication was applied to the control group, UCB (50 μM) and caffeine (100 μM) were applied to the bilirubin and caffeine groups for 24 h. Prophylactic and therapeutic caffeine groups were treated with caffeine 4 h before and after UCB exposure. The effects of caffeine were investigated in rat astrocytes exposed to UCB in terms of cell viability, apoptosis, antioxidant defense, proinflammatory cytokines, and Toll-like receptor (TLR)s.
Compared to the control group, UCB increased apoptosis, malondialdehyde (MDA), tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-6, total nitrate/nitrite, and TLR4 levels, and decreased cell viability, catalase (CAT), glutathione peroxidase (GPx), superoxide dismutase (SOD) activities, glutathione, and TLR9 levels (for all p < .001). Conversely, prophylactic and therapeutic caffeine improved the detrimental effects of UCB.
Caffeine seems encouraging for the prevention and treatment of bilirubin neurotoxicity in rats by means of its antiapoptotic, antioxidant, anti-inflammatory, anti-nitrosative, and anti-TLR-4 properties.
由于尿苷二磷酸葡萄糖醛酸基转移酶1A1（UGT1A1）不成熟导致胆红素在脑内蓄积，未结合胆红素（UCB）可能会引起早产儿神经毒性。据报道，用于治疗早产儿呼吸暂停的咖啡因可减少极低出生体重儿的机械通气需求、支气管肺发育不良、动脉导管未闭、脑瘫和神经发育障碍的发生率。然而，咖啡因对高胆红素血症的影响尚未阐明。
我们采用改良的Cole和de Vellis方法，从2日龄Wistar白化大鼠获取星形胶质细胞培养物。实验中使用对50%星形胶质细胞有毒的UCB浓度以及使细胞活力提高100%的咖啡因浓度。对照组不施加任何药物，胆红素组和咖啡因组分别施加UCB（50μM）和咖啡因（100μM）24小时。预防性咖啡因组和治疗性咖啡因组在UCB暴露前4小时和暴露后进行咖啡因处理。从细胞活力、凋亡、抗氧化防御、促炎细胞因子和Toll样受体（TLR）方面，研究咖啡因对暴露于UCB的大鼠星形胶质细胞的影响。
与对照组相比，UCB增加了凋亡、丙二醛（MDA）、肿瘤坏死因子-α（TNF-α）、白细胞介素（IL）-1β、IL-6、总硝酸盐/亚硝酸盐和TLR4水平，降低了细胞活力、过氧化氢酶（CAT）、谷胱甘肽过氧化物酶（GPx）、超氧化物歧化酶（SOD）活性、谷胱甘肽和TLR9水平（所有p < 0.001）。相反，预防性和治疗性咖啡因改善了UCB的有害影响。
咖啡因因其抗凋亡、抗氧化、抗炎、抗亚硝化和抗TLR-4特性，似乎对预防和治疗大鼠胆红素神经毒性具有积极作用。
