Each year, 1 million children die due to perinatal asphyxia; however, there are no effective drugs to protect the neonatal brain against hypoxic/ischemic damage. In this study, we demonstrated for the first time the neuroprotective capacity of 3,3’-diindolylmethane (DIM) in an in vivo model of rat perinatal asphyxia, which has translational value and corresponds to hypoxic/ischemic episodes in human newborns. Posttreatment with DIM restored the weight of the ipsilateral hemisphere and normalized cell number in the brain structures of rats exposed to perinatal asphyxia. DIM also downregulated the mRNA expression of HIF1A-regulated Bnip3 and Hif1a which is a hypoxic marker, and the expression of miR-181b which is an indicator of perinatal asphyxia. In addition, DIM inhibited apoptosis and oxidative stress accompanying perinatal asphyxia through: downregulation of FAS, CASP-3, CAPN1, GPx3 and SOD-1, attenuation of caspase-9 activity, and upregulation of anti-apoptotic Bcl2 mRNA. The protective effects of DIM were accompanied by the inhibition of the AhR and NMDA signaling pathways, as indicated by the reduced expression levels of AhR, ARNT, CYP1A1, GluN1 and GluN2B, which was correlated with enhanced global DNA methylation and the methylation of the Ahr and Grin2b genes. Because our study provided evidence that in rat brain undergoing perinatal asphyxia, DIM predominantly targets AhR and NMDA, we postulate that compounds that possess the ability to inhibit their signaling are promising therapeutic tools to prevent stroke.

每年有 100 万儿童因围产期窒息而死亡；然而，没有有效的药物来保护新生儿大脑免受缺氧/缺血性损伤。在这项研究中，我们首次证明了 3,3'-二吲哚基甲烷 (DIM) 在大鼠围产期窒息体内模型中的神经保护能力，该模型具有转化价值，对应于人类新生儿的缺氧/缺血发作。用昏暗的后处理恢复了同侧半球的重量，并使暴露于围产期窒息的大鼠大脑结构中的细胞数量恢复正常。DIM 还下调了 HIF1A 调节的Bnip3和Hif1a的 mRNA 表达这是一种缺氧标志物，以及作为围产期窒息指标的 miR-181b 的表达。此外，DIM 通过以下方式抑制伴随围产期窒息的细胞凋亡和氧化应激：FAS、CASP-3、CAPN1、GPx3 和 SOD-1 的下调，caspase-9 活性的减弱，以及抗凋亡Bcl2 mRNA 的上调。DIM 的保护作用伴随着 AhR 和 NMDA 信号通路的抑制，表现为 AhR、ARNT、CYP1A1、GluN1 和 GluN2B 的表达水平降低，这与增强的整体 DNA 甲基化和Ahr的甲基化相关和Grin2b基因。因为我们的研究提供的证据表明，在经历围产期窒息的大鼠大脑中，DIM 主要针对 AhR 和 NMDA，我们假设具有抑制其信号传导能力的化合物是预防中风的有希望的治疗工具。