Department of Physiology & Pharmacology, Loma Linda University School of Medicine, Loma Linda, CA, USA.
Department of Physiology & Pharmacology, Loma Linda University School of Medicine, Loma Linda, CA, USA; Departments of Anesthesiology and Neurosurgery, Loma Linda University School of Medicine, Loma Linda, CA, USA.
Exp Neurol. 2023 Jan;359:114257. doi: 10.1016/j.expneurol.2022.114257. Epub 2022 Oct 21.
Germinal matrix hemorrhage (GMH) is one of the leading causes of morbidity and mortality in preterm infants in the United States, with little progress made in its clinical management. Blood clots disrupting normal cerebrospinal fluid circulation and absorption after germinal matrix hemorrhage are key contributors towards post-hemorrhagic hydrocephalus development. n-formyl peptide receptor 2 (FPR2), a G-protein-coupled receptor, has been associated with the activation of p-ERK1/2, which in turn promotes the transcription of the DUSP1 gene, which may play a role in CD36 signaling. CD36 scavenger, a transmembrane glycoprotein, plays an essential role in microglia phagocytic blood clot clearance after GMH. FPR2's role in blood clot clearance after hemorrhagic stroke is unknown. We hypothesize that FPR2 activation by FPR2 agonist Annexin A1 (AnxA1) will enhance hematoma resolution via the upregulation of the CD36 signaling pathway, thereby improving short- and long-term neurological outcomes. Bacterial collagenase (0.3 U) was infused intraparenchymally into the right hemispheric ganglionic eminence in P7 rat pups to induce GMH. AnxA1 and FPR2 Inhibitor (Boc2) were given at 1-h post-GMH via intranasal administration. FPR2 CRISPR was given 48-h prior to GMH induction. Short-term neurological deficits were assessed using negative geotaxis test. Hematoma volume was assessed using hemoglobin assay. Protein expression was assessed using western blots. Long-term neurocognitive deficits and motor coordination were assessed using Morris water maze, rotarod, and foot fault tests. We have demonstrated that AnxA1 treatment enhances hematoma resolution and improved short and long-term outcomes. Lastly, FPR2 agonist AnxA1 treatment resulted in the upregulation of the FPR2/p-ERK(1/2)/DUSP1/CD36 signaling pathway.
脑室内出血(GMH)是美国早产儿发病率和死亡率的主要原因之一,但在其临床管理方面几乎没有进展。GMH 后,阻断正常脑脊液循环和吸收的血凝块是导致出血后脑积水发展的关键因素。形式基肽受体 2(FPR2)是一种 G 蛋白偶联受体,与 p-ERK1/2 的激活有关,p-ERK1/2 又促进 DUSP1 基因的转录,该基因可能在 CD36 信号传导中发挥作用。CD36 清道夫,一种跨膜糖蛋白,在 GMH 后小胶质细胞吞噬血凝块清除中起着至关重要的作用。FPR2 在出血性中风后血凝块清除中的作用尚不清楚。我们假设 FPR2 激动剂 Annexin A1(AnxA1)通过激活 FPR2 来增强血肿的溶解,从而上调 CD36 信号通路,从而改善短期和长期神经预后。在 P7 大鼠幼仔的右神经节隆起内脑实质内注入细菌胶原酶(0.3 U)以诱导 GMH。在 GMH 后 1 小时通过鼻内给药给予 AnxA1 和 FPR2 抑制剂(Boc2)。FPR2 CRISPR 在 GMH 诱导前 48 小时给予。使用负趋地性试验评估短期神经功能缺损。使用血红蛋白测定法评估血肿体积。使用 Western blot 评估蛋白质表达。使用 Morris 水迷宫,转棒和足失误测试评估长期神经认知缺陷和运动协调能力。我们已经证明,AnxA1 治疗可增强血肿溶解并改善短期和长期结果。最后,FPR2 激动剂 AnxA1 治疗导致 FPR2/p-ERK(1/2)/DUSP1/CD36 信号通路的上调。