Jing Jie, Chen Shiling, Wu Xuan, Yang Jingfei, Liu Xia, Wang Jiahui, Wang Jingyi, Li Yunjie, Zhang Ping, Tang Zhouping
Department of Neurology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei Province, China.
Department of Neurology, Qilu Hospital of Shandong University, Jinan, Shandong Province, China.
Neural Regen Res. 2024 Jul 29. doi: 10.4103/NRR.NRR-D-23-01953.
Recombinant tissue plasminogen activator is commonly used for hematoma evacuation in minimally invasive surgery following intracerebral hemorrhage. However, during minimally invasive surgery, recombinant tissue plasminogen activator may come into contact with brain tissue. Therefore, a thorough assessment of its safety is required. In this study, we established a mouse model of intracerebral hemorrhage induced by type VII collagenase. We observed that the administration of recombinant tissue plasminogen activator without hematoma aspiration significantly improved the neurological function of mice with intracerebral hemorrhage, reduced pathological damage, and lowered the levels of apoptosis and autophagy in the tissue surrounding the hematoma. In an in vitro model of intracerebral hemorrhage using primary cortical neurons induced by hemin, the administration of recombinant tissue plasminogen activator suppressed neuronal apoptosis, autophagy, and endoplasmic reticulum stress. Transcriptome sequencing analysis revealed that recombinant tissue plasminogen activator upregulated the phosphoinositide 3-kinase/RAC-alpha serine/threonine-protein kinase/mammalian target of rapamycin pathway in neurons. Moreover, the phosphoinositide 3-kinase inhibitor LY294002 abrogated the neuroprotective effects of recombinant tissue plasminogen activator in inhibiting excessive apoptosis, autophagy, and endoplasmic reticulum stress. Furthermore, to specify the domain of recombinant tissue plasminogen activator responsible for its neuroprotective effects, various inhibitors were used to target distinct domains. It has been revealed that the epidermal growth factor receptor inhibitor AG-1478 reversed the effect of recombinant tissue plasminogen activator on the phosphoinositide 3-kinase/RAC-alpha serine/threonine-protein kinase/mammalian target of rapamycin pathway. These findings suggest that recombinant tissue plasminogen activator exerts a direct neuroprotective effect on neurons following intracerebral hemorrhage, possibly through activation of the phosphoinositide 3-kinase/RAC-alpha serine/threonine-protein kinase/mammalian target of rapamycin pathway.
重组组织型纤溶酶原激活剂常用于脑出血后微创手术中的血肿清除。然而，在微创手术过程中，重组组织型纤溶酶原激活剂可能会与脑组织接触。因此，需要对其安全性进行全面评估。在本研究中，我们建立了VII型胶原酶诱导的脑出血小鼠模型。我们观察到，在不进行血肿抽吸的情况下给予重组组织型纤溶酶原激活剂可显著改善脑出血小鼠的神经功能，减少病理损伤，并降低血肿周围组织中的凋亡和自噬水平。在使用血红素诱导的原代皮质神经元建立的脑出血体外模型中，给予重组组织型纤溶酶原激活剂可抑制神经元凋亡、自噬和内质网应激。转录组测序分析显示，重组组织型纤溶酶原激活剂上调了神经元中的磷脂酰肌醇3激酶/RAC-α丝氨酸/苏氨酸蛋白激酶/雷帕霉素哺乳动物靶标通路。此外，磷脂酰肌醇3激酶抑制剂LY294002消除了重组组织型纤溶酶原激活剂在抑制过度凋亡、自噬和内质网应激方面的神经保护作用。此外，为了明确重组组织型纤溶酶原激活剂发挥神经保护作用的结构域，使用了各种抑制剂靶向不同结构域。结果显示，表皮生长因子受体抑制剂AG-1478逆转了重组组织型纤溶酶原激活剂对磷脂酰肌醇3激酶/RAC-α丝氨酸/苏氨酸蛋白激酶/雷帕霉素哺乳动物靶标通路的作用。这些发现表明，重组组织型纤溶酶原激活剂在脑出血后对神经元发挥直接神经保护作用，可能是通过激活磷脂酰肌醇3激酶/RAC-α丝氨酸/苏氨酸蛋白激酶/雷帕霉素哺乳动物靶标通路实现的。
