Anxiety is reportedly one of the most common mental changes after traumatic brain injury (TBI). Perineuronal nets (PNNs) produced by astrocytes in the lateral hypothalamus (LHA) that surround gamma-aminobutyric acid-ergic (GABAergic) neurons have been associated with anxiety. The potent anti-tumor effects of Spautin-1, a novel autophagy inhibitor, have been documented in malignant melanoma; moreover, the inhibition of autophagy is reported to mitigate anxiety disorders. However, little is known about the ability of spautin-1 to alleviate anxiety. In this study, we sought to investigate whether spautin-1 could alleviate anxiety-like behaviors post-TBI by reducing the loss of PNNs in the LHA. A mild TBI was established in mice through Feeney’s weight-drop model. Then, Spautin-1 (20 mmol/2 μl) was immediately administered into the left lateral ventricle. Behavioral and pathological changes were assessed at 24 h, 7 days, 30 days, 31 days and 32 days after TBI by the neurological severity scores (NSS), open field test (OFT), elevated plus-maze (EPM) test, western blot, immunofluorescence assays and electron microscopy. Spautin-1 significantly reversed TBI-induced decreased time in the central zone during OFT and in the open-arm during the EPM test. Spautin-1 also increased PNNs around GABAergic neurons indicated by WFA- plus GAD2- positive A2-type astrocytes and attenuated M1-type microglia in the LHA 32 days after TBI compared to TBI alone. Moreover, compared to mice that only underwent TBI, spautin-1 downregulated autophagic vacuoles, abnormal organelles, the expression of Beclin 1, USP13, phospho-TBK1, and phospho-IRF3 and upregulated the levels of cleaved caspase-3, -7 and -9, but failed to increase TUNEL-positive cells in the LHA at 24 h. Spautin-1 alleviated anxiety-like behavior in mice exposed to mild TBI; this protective mechanism may be associated with decreased PNNs loss around GABAergic neurons via immunologically silent apoptosis induced by the caspase cascade.

据报道，焦虑是创伤性脑损伤（TBI）后最常见的精神变化之一。由下丘脑外侧 (LHA) 中的星形胶质细胞产生的围绕伽马氨基丁酸能 (GABA 能) 神经元的神经周围网 (PNN) 与焦虑有关。Spautin-1（一种新型自噬抑制剂）的有效抗肿瘤作用已在恶性黑色素瘤中得到证实；此外，据报道，抑制自噬可以减轻焦虑症。然而，人们对 spautin-1 缓解焦虑的能力知之甚少。在本研究中，我们试图研究 spautin-1 是否可以通过减少 LHA 中 PNN 的丢失来减轻 TBI 后的焦虑样行为。通过菲尼的体重下降模型，在小鼠身上建立了轻度脑损伤。然后，立即将Spautin-1（20mmol/2μl）注入左侧侧脑室。通过神经严重程度评分（NSS）、旷场试验（OFT）、高架十字迷宫（EPM）试验、蛋白质印迹评估TBI后24小时、7天、30天、31天和32天的行为和病理变化。 、免疫荧光测定和电子显微镜。Spautin-1 显着逆转了 TBI 引起的 OFT 中心区时间缩短和 EPM 测试期间开臂时间缩短。与单独的 TBI 相比，在 TBI 后 32 天，Spautin-1 还增加了 GABA 能神经元周围的 PNN（由 WFA 加 GAD2 阳性的 A2 型星形胶质细胞表明），并减弱了 LHA 中的 M1 型小胶质细胞。此外，与仅接受TBI的小鼠相比，spautin-1下调自噬泡、异常细胞器、Beclin 1、USP13、磷酸-TBK1和磷酸-IRF3的表达，并上调cleaved caspase-3、-7和-的水平。 9，但未能在24小时增加LHA中的TUNEL阳性细胞。Spautin-1 缓解了轻度 TBI 小鼠的焦虑样行为；这种保护机制可能与通过 caspase 级联诱导的免疫沉默细胞凋亡减少 GABA 能神经元周围的 PNN 丢失有关。