MicroRNA-181c provides neuroprotection in an intracerebral hemorrhage model
MicroRNA-181c provides neuroprotection in an intracerebral hemorrhage model
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DOI：
10.4103/1673-5374.272612
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发表时间：
2020-07-01
影响因子：
6.1
通讯作者：
He, Zhi-Yi
中科院分区：
文献类型：
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作者：
Lu, Xi;Zhang, Hui-Yuan;He, Zhi-Yi
细胞凋亡是脑出血早期的重要因素。 MiR-181c 在细胞凋亡中发挥关键的调节作用。然而，miR-181c是否参与脑出血后早期细胞凋亡尚不清楚。因此，进行了体外和体内实验来检验这一假设。体内实验：将VII型胶原酶注射到成年Sprague-Dawley大鼠基底节区，建立脑出血模型。脑出血后4小时原位注射MiR-181c模拟物或抑制剂。模型建立后1、7和14天评估神经功能缺陷（神经严重程度评分）。模型建立14天后进行末端脱氧核苷酸转移酶介导的脱氧尿苷三磷酸缺口末端标记和蛋白质印迹测定。体外实验：缺氧缺糖条件下培养PC12细胞，加入氯化血红素体外模拟脑出血。添加 miR-181c 模拟物或抑制剂来调节 miR-181c 表达。进行了3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴化物测定、荧光素酶报告系统和蛋白质印迹测定。实验结果显示脑出血患者和大鼠脑组织中miR-181c的表达存在差异。此外，体外实验发现，miR-181c过表达可上调Bcl-2/Bax比值从而抑制细胞凋亡，而抑制miR-181c表达可降低Bcl-2/Bax比值，加剧细胞凋亡。通过磷酸肌醇 3 激酶 (PI3K)/Akt 途径通过靶向十号染色体上缺失的磷酸酶和张力蛋白同源物 (PTEN) 来调节细胞凋亡。较高的 miR-181c 过度表达与较低的神经系统严重程度评分相关，表明神经功能恢复得更好。总之，miR-181c通过调节细胞凋亡影响脑出血的预后，并且这些作用可能是通过靶向PTEN\PI3K/Akt通路和Bcl-2/Bax比值直接介导和调节的。此外，这些结果表明miR-181c通过调节神经细胞凋亡在脑出血中发挥神经保护作用，从而为脑出血的预防和治疗提供了潜在的靶点。人血清检测于2012年2月20日获得中国医科大学伦理委员会授权（批准号：2012-38-1）。该方案已在中国临床试验注册中心注册（注册号：ChiCTR-COC-17013559）。该动物研究于2017年3月8日获得中国医科大学机构动物保育与使用委员会批准（批准号：2017008）。
Apoptosis is an important factor during the early stage of intracerebral hemorrhage. MiR-181c plays a key regulatory role in apoptosis. However, whether miR-181c is involved in apoptosis of prophase cells after intracerebral hemorrhage remains unclear. Therefore, in vitro and in vivo experiments were conducted to test this hypothesis. In vivo experiments: collagenase type VII was injected into the basal ganglia of adult Sprague-Dawley rats to establish an intracerebral hemorrhage model. MiR-181c mimic or inhibitor was injected in situ 4 hours after intracerebral hemorrhage. Neurological functional defects (neurological severity scores) were assessed 1, 7, and 14 days after model establishment. Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling and western blot assay were conducted 14 days after model establishment. In vitro experiments: PC12 cells were cultured under oxygen-glucose deprivation, and hemins were added to simulate intracerebral hemorrhage in vitro. MiR-181c mimic or inhibitor was added to regulate miR-181c expression. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, luciferase reporter system, and western blot assay were performed. Experimental results revealed differences in miR-181c expression in brain tissues of both patients and rats with cerebral hemorrhage. In addition, in vitro experiments found that miR-181c overexpression could upregulate the Bcl-2/Bax ratio to inhibit apoptosis, while inhibition of miR-181c expression could reduce the Bcl-2/Bax ratio and aggravate apoptosis of cells. Regulation of apoptosis occurred through the phosphoinositide 3 kinase (PI3K)/Akt pathway by targeting of phosphatase and tensin homolog deleted on chromosome ten (PTEN). Higher miR-181c overexpression correlated with lower neurological severity scores, indicating better recovery of neurological function. In conclusion, miR-181c affects the prognosis of intracerebral hemorrhage by regulating apoptosis, and these effects might be directly mediated and regulated by targeting of the PTEN\PI3K/Akt pathway and Bcl-2/Bax ratio. Furthermore, these results indicated that miR-181c played a neuroprotective role in intracerebral hemorrhage by regulating apoptosis of nerve cells, thus providing a potential target for the prevention and treatment of intracerebral hemorrhage. Testing of human serum was authorized by the Ethics Committee of China Medical University (No. 2012-38-1) on February 20, 2012. The protocol was registered with the Chinese Clinical Trial Registry (Registration No. ChiCTR-COC-17013559). The animal study was approved by the Institutional Animal Care and Use Committee of China Medical University (approval No. 2017008) on March 8, 2017.