Inhibiting endogenous tissue plasminogen activator enhanced neuronal apoptosis and axonal injury after traumatic brain injury

Jun-Jie Zhao; Zun-Wei Liu; Bo Wang; Ting-Qin Huang; Dan Guo; Yong-Lin Zhao; Jin-Ning Song

doi:10.4103/1673-5374.266914

Inhibiting endogenous tissue plasminogen activator enhanced neuronal apoptosis and axonal injury after traumatic brain injury

Neural Regen Res. 2020 Apr;15(4):667-675. doi: 10.4103/1673-5374.266914.

Authors

Jun-Jie Zhao¹, Zun-Wei Liu², Bo Wang¹, Ting-Qin Huang³, Dan Guo⁴, Yong-Lin Zhao⁵, Jin-Ning Song¹

Affiliations

¹ Department of Neurosurgery, the First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi Province, China.
² Institute of Organ Transplantation, the First Affiliated Hospital of Xi'an Jiaotong University; Department of Renal Transplantation, Nephropathy Hospital, the First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi Province, China.
³ Department of Neurosurgery, the Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi Province, China.
⁴ Department of Science and Technology, Xi'an Medical University, Xi'an, Shaanxi Province, China.
⁵ Department of Oncology, the Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi Province, China.

Abstract

Tissue plasminogen activator is usually used for the treatment of acute ischemic stroke, but the role of endogenous tissue plasminogen activator in traumatic brain injury has been rarely reported. A rat model of traumatic brain injury was established by weight-drop method. The tissue plasminogen activator inhibitor neuroserpin (5 μL, 0.25 mg/mL) was injected into the lateral ventricle. Neurological function was assessed by neurological severity score. Neuronal and axonal injuries were assessed by hematoxylin-eosin staining and Bielschowsky silver staining. Protein level of endogenous tissue plasminogen activator was analyzed by western blot assay. Apoptotic marker cleaved caspase-3, neuronal marker neurofilament light chain, astrocyte marker glial fibrillary acidic protein and microglial marker Iba-1 were analyzed by immunohistochemical staining. Apoptotic cell types were detected by immunofluorescence double labeling. Apoptotic cells in the damaged cortex were detected by terminal deoxynucleotidyl transferase-mediated digoxigenin-dUTP-biotin nick-end labeling staining. Degenerating neurons in the damaged cortex were detected by Fluoro-Jade B staining. Expression of tissue plasminogen activator was increased at 6 hours, and peaked at 3 days after traumatic brain injury. Neuronal apoptosis and axonal injury were detected after traumatic brain injury. Moreover, neuroserpin enhanced neuronal apoptosis, neuronal injury and axonal injury, and activated microglia and astrocytes. Neuroserpin further deteriorated neurobehavioral function in rats with traumatic brain injury. Our findings confirm that inhibition of endogenous tissue plasminogen activator aggravates neuronal apoptosis and axonal injury after traumatic brain injury, and activates microglia and astrocytes. This study was approved by the Biomedical Ethics Committee of Animal Experiments of Shaanxi Province of China in June 2015.

Keywords: apoptosis; astrocytes; axonal injury; inflammation; microglia; nerve regeneration; neural regeneration; neuronal injury; neurons; neuroserpin; tissue plasminogen activator; traumatic brain injury.