Hemorrhagic transformation can complicate ischemic strokes after recanalization treatment within a time window that requires early intervention. To determine potential therapeutic effects of matrilin-3, rat cerebral ischemia-reperfusion was produced using transient middle cerebral artery occlusion (tMCAO); intracranial hemorrhage and infarct volumes were assayed through hemoglobin determination and 2,3,5-triphenyltetrazoliumchloride (TTC) staining, respectively. Oxygen-glucose deprivation (OGD) modeling of ischemia was performed on C8-D1A cells. Interactions between matrilin-3 and YTH N6-methyladenosine RNA binding protein F2 (YTHDF2) were determined using RNA immunoprecipitation assay and actinomycin D treatment. Reperfusion after tMCAO modeling increased hemorrhage, hemoglobin content, and infarct volumes; these were alleviated by matrilin treatment. Matrilin-3 was expressed at low levels and YTHDF2 was expressed at high levels in ischemic brains. In OGD-induced cells, matrilin-3 was negatively regulated by YTHDF2. Matrilin-3 overexpression downregulated p-PI3K/PI3K, p-AKT/AKT, ZO-1, VE-cadherin and occludin, and upregulated p-JNK/JNK in ischemic rat brains; these effects were reversed by LY294002 (a PI3K inhibitor). YTHDF2 knockdown inactivated the PI3K/AKT pathway, inhibited inflammation and decreased blood-brain barrier-related protein levels in cells; these effects were reversed by matrilin-3 deficiency. These results indicate that YTHDF2-regulated matrilin-3 protected ischemic rats against post-reperfusion hemorrhagic transformation via the PI3K/AKT pathway and that matrilin may have therapeutic potential in ischemic stroke.

中文翻译：

---

YTHDF2 调节的 matrilin-3 通过 PI3K/AKT 途径减轻缺血性中风再灌注后出血性转化

在需要早期干预的时间窗口内进行再通治疗后，出血性转化可能会使缺血性中风复杂化。为了确定 matrilin-3 的潜在治疗效果，使用短暂性大脑中动脉闭塞 (tMCAO) 产生大鼠脑缺血再灌注；分别通过血红蛋白测定和2,3,5-三苯基氯化四唑（TTC）染色来测定颅内出血和梗塞体积。对 C8-D1A 细胞进行缺氧葡萄糖剥夺 (OGD) 缺血模型。使用 RNA 免疫沉淀测定和放线菌素 D 处理测定 matrilin-3 和 YTH N6-甲基腺苷 RNA 结合蛋白 F2 (YTHDF2) 之间的相互作用。tMCAO 建模后的再灌注增加了出血量、血红蛋白含量和梗塞体积；这些通过母体素治疗得到缓解。在缺血脑中，Matrilin-3 低水平表达，YTHDF2 高水平表达。在 OGD 诱导的细胞中，matrilin-3 受到 YTHDF2 的负调节。在缺血大鼠脑中，Matrilin-3 过表达下调 p-PI3K/PI3K、p-AKT/AKT、ZO-1、VE-cadherin 和 occludin，上调 p-JNK/JNK；这些效应可被 LY294002（一种 PI3K 抑制剂）逆转。YTHDF2敲低使PI3K/AKT通路失活，抑制炎症并降低细胞中血脑屏障相关蛋白水平；这些效应因 matrilin-3 缺乏而逆转。这些结果表明，YTHDF2调节的matrilin-3通过PI3K/AKT途径保护缺血大鼠免受再灌注后出血性转化，并且matrilin可能具有治疗缺血性中风的潜力。