Pandya Jignesh D, Musyaju Sudeep, Modi Hiren R, Cao Ying, Flerlage William J, Huynh Linda, Kociuba Brittany, Visavadiya Nishant P, Kobeissy Firas, Wang Kevin, Gilsdorf Janice S, Scultetus Anke H, Shear Deborah A
Brain Trauma Neuroprotection (BTN) Branch, Walter Reed Army Institute of Research (WRAIR), Silver Spring, MD, 20910, USA.
Brain Trauma Neuroprotection (BTN) Branch, Walter Reed Army Institute of Research (WRAIR), Silver Spring, MD, 20910, USA.
Free Radic Biol Med. 2023 Mar;198:44-58. doi: 10.1016/j.freeradbiomed.2023.02.001. Epub 2023 Feb 7.
Traumatic Brain Injury (TBI) is caused by the external physical assaults damages the brain. It is a heterogeneous disorder that remains a leading cause of death and disability in the military and civilian population of the United States. Preclinical investigations of mitochondrial responses in TBI have ascertained that mitochondrial dysfunction is an acute indicator of cellular damage and plays a pivotal role in long-term injury progression through cellular excitotoxicity. The current study was designed to provide an in-depth evaluation of mitochondrial endpoints with respect to redox and calcium homeostasis, and cell death responses following penetrating TBI (PTBI). To evaluate these pathological cascades, anesthetized adult male rats (N = 6/group) were subjected to either 10% unilateral PTBI or Sham craniectomy. Animals were euthanized at 24 h post-PTBI, and purified mitochondrial fractions were isolated from the brain injury core and perilesional areas. Overall, increased reactive oxygen and nitrogen species (ROS/RNS) production, and elevated oxidative stress markers such as 4-hydroxynonenal (4-HNE), 3-nitrotyrosine (3-NT), and protein carbonyls (PC) were observed in the PTBI group compared to Sham. Mitochondrial antioxidants such as glutathione, peroxiredoxin (PRX-3), thioredoxin (TRX), nicotinamide adenine dinucleotide phosphate (NADPH), superoxide dismutase (SOD), and catalase (CAT) levels were significantly decreased after PTBI. Likewise, PTBI mitochondria displayed significant loss of Ca homeostasis, early opening of mitochondrial permeability transition pore (mPTP), and increased mitochondrial swelling. Both, outer and inner mitochondrial membrane integrity markers, such as voltage-dependent anion channels (VDAC) and cytochrome c (Cyt C) expression were significantly decreased following PTBI. The apoptotic cell death was evidenced by significantly decreased B-cell lymphoma-2 (Bcl-2) and increased glyceraldehyde 3-phosphate dehydrogenase (GAPDH) expression after PTBI. Collectively, current results highlight the comprehensive picture of mitochondria-centric acute pathophysiological responses following PTBI, which may be utilized as novel prognostic indicators of disease progression and theragnostic indicators for evaluating neuroprotection therapeutics following TBI.
创伤性脑损伤（TBI）是由外部物理攻击导致大脑受损引起的。它是一种异质性疾病，在美国军人和平民中仍然是死亡和残疾的主要原因。TBI中线粒体反应的临床前研究已确定，线粒体功能障碍是细胞损伤的急性指标，并通过细胞兴奋性毒性在长期损伤进展中起关键作用。本研究旨在深入评估穿透性TBI（PTBI）后线粒体在氧化还原和钙稳态以及细胞死亡反应方面的终点指标。为了评估这些病理级联反应，将麻醉的成年雄性大鼠（每组N = 6只）进行10%单侧PTBI或假颅骨切除术。动物在PTBI后24小时安乐死，从脑损伤核心和损伤周围区域分离纯化的线粒体部分。总体而言，与假手术组相比，PTBI组观察到活性氧和氮物种（ROS/RNS）产生增加，以及氧化应激标志物如4-羟基壬烯醛（4-HNE）、3-硝基酪氨酸（3-NT）和蛋白质羰基（PC）升高。PTBI后线粒体抗氧化剂如谷胱甘肽、过氧化物酶（PRX-3）、硫氧还蛋白（TRX）、烟酰胺腺嘌呤二核苷酸磷酸（NADPH）、超氧化物歧化酶（SOD）和过氧化氢酶（CAT）水平显著降低。同样，PTBI线粒体显示钙稳态显著丧失，线粒体通透性转换孔（mPTP）早期开放，线粒体肿胀增加。PTBI后，线粒体外膜和内膜完整性标志物如电压依赖性阴离子通道（VDAC）和细胞色素c（Cyt C）表达均显著降低。PTBI后B细胞淋巴瘤-2（Bcl-2）显著降低和甘油醛-3-磷酸脱氢酶（GAPDH）表达增加证明了凋亡性细胞死亡。总的来说，目前的结果突出了PTBI后以线粒体为中心的急性病理生理反应的全貌，这可能被用作疾病进展的新型预后指标和评估TBI后神经保护治疗的治疗诊断指标。
