Rey-Funes Manuel, Contartese Daniela S, Peláez Rafael, García-Sanmartín Josune, Narro-Íñiguez Judit, Soliño Manuel, Fernández Juan Carlos, Sarotto Aníbal, Ciranna Nicolás S, López-Costa Juan José, Dorfman Verónica B, Larrayoz Ignacio M, Loidl C Fabián, Martínez Alfredo
Laboratorio de Neuropatología Experimental, Instituto de Biología Celular y Neurociencia "Prof, E. De Robertis" (IBCN), Facultad de Medicina, CONICET - Universidad de Buenos Aires, Buenos Aires, Argentina.
Departamento de Biología Celular, Histología, Embriología y Genética, Instituto de Biología Celular y Neurociencia "Prof, E. De Robertis" (IBCN), Facultad de Medicina, Universidad de Buenos Aires, Buenos Aires, Argentina.
Front Pharmacol. 2021 Apr 8;12:651599. doi: 10.3389/fphar.2021.651599. eCollection 2021.
Perinatal asphyxia (PA) can cause retinopathy and different degrees of visual loss, including total blindness. In a rat model of PA, we have previously shown a protective effect of hypothermia on the retina when applied simultaneously with the hypoxic insult. In the present work, we evaluated the possible protective effect of hypothermia on the retina of PA rats when applied immediately after delivery. Four experimental groups were studied: Rats born naturally as controls (CTL), animals that were exposed to PA for 20 min at 37°C (PA), animals exposed to PA for 20 min at 15°C (HYP), and animals that were exposed to PA for 20 min at 37°C and, immediately after birth, kept for 15 min at 8°C (HYP-PA). To evaluate the integrity of the visual pathway, animals were subjected to electroretinography at 45 days of age. Molecular (real time PCR) and histological (immunohistochemistry, immunofluorescence, TUNEL assay) techniques were applied to the eyes of all experimental groups collected at 6, 12, 24, and 48 h, and 6 days after birth. PA resulted in a significant reduction in the amplitude of the a- and b-wave and oscillatory potentials (OP) of the electroretinogram. All animals treated with hypothermia had a significant correction of the a-wave and OP, but the b-wave was fully corrected in the HYP group but only partially in the HYP-PA group. The number of TUNEL-positive cells increased sharply in the ganglion cell layer of the PA animals and this increase was significantly prevented by both hypothermia treatments. Expression of the cold-shock proteins, cold-inducible RNA binding protein (CIRP) and RNA binding motif protein 3 (RBM3), was undetectable in retinas of the CTL and PA groups, but they were highly expressed in ganglion neurons and cells of the inner nuclear layer of the HYP and HYP-PA groups. In conclusion, our results suggest that a post-partum hypothermic shock could represent a useful and affordable method to prevent asphyxia-related vision disabling sequelae.
围产期窒息(PA)可导致视网膜病变和不同程度的视力丧失,包括完全失明。在PA大鼠模型中,我们之前已表明,在缺氧损伤同时应用低温对视网膜具有保护作用。在本研究中,我们评估了产后立即应用低温对PA大鼠视网膜可能产生的保护作用。研究了四个实验组:自然出生的大鼠作为对照组(CTL);在37°C下暴露于PA 20分钟的动物(PA);在15°C下暴露于PA 20分钟的动物(HYP);以及在37°C下暴露于PA 20分钟且出生后立即在8°C下保持15分钟的动物(HYP-PA)。为评估视觉通路的完整性,在动物45日龄时进行视网膜电图检查。对所有实验组出生后6、12、24和48小时以及6天收集的眼睛应用分子(实时PCR)和组织学(免疫组织化学、免疫荧光、TUNEL检测)技术。PA导致视网膜电图的a波和b波振幅以及振荡电位(OP)显著降低。所有接受低温治疗的动物的a波和OP均有显著改善,但HYP组的b波完全恢复,而HYP-PA组仅部分恢复。PA动物神经节细胞层中TUNEL阳性细胞数量急剧增加,两种低温治疗均显著抑制了这种增加。冷休克蛋白、冷诱导RNA结合蛋白(CIRP)和RNA结合基序蛋白3(RBM3)在CTL组和PA组的视网膜中未检测到表达,但在HYP组和HYP-PA组的神经节神经元和内核层细胞中高表达。总之,我们的结果表明,产后低温休克可能是一种预防窒息相关视力致残后遗症的有用且经济的方法。