Adenoviral transfer of hemopexin gene attenuates oxidative stress and apoptosis in cultured primary cortical neuron cell exposed to blood clot
Adenoviral transfer of hemopexin gene attenuates oxidative stress and apoptosis in cultured primary cortical neuron cell exposed to blood clot
复制标题
血红素基因的腺病毒转移可减轻暴露于血凝块的培养原代皮质神经元细胞的氧化应激和细胞凋亡
DOI：
10.1097/wnr.0000000000001510
复制
发表时间：
2020-10-14
期刊：
影响因子：
1.7
通讯作者：
Chen，Lifen
中科院分区：
文献类型：
--
作者：
Liu，Yi;Tan，Changhong;Chen，Lifen
越来越多的实验证据表明，从血红素中释放的氯化血红素是一种强氧化剂，并在颅内血肿中蓄积。血红素结合素（Hpx）减少氯化血红素的积累和神经细胞的催化作用。在前期的研究中，我们观察到Hpx基因敲除可加重脑出血小鼠的纹状体损伤和行为缺陷。目的探讨血凝素（Hpx）对血凝块神经细胞氧化损伤和凋亡的影响。方法用腺病毒Hpx基因转染神经元和胶质细胞。使用插入transwell将转染的原代神经元-神经胶质细胞与50 μl动脉血凝块共培养。假手术组与含血清28 μl的DMEM/F12共培养50 μl，对照组转染腺病毒载体。分别于12、24 h检测丙二醛（MDA）、超氧化物歧化酶（SOD）、谷胱甘肽（GSH）、细胞凋亡、HO-1和caspase-3的表达。结果Hpx组MDA含量降低（P < 0.01），SOD和GSH含量升高（分别为P < 0.05和P < 0.01）。流式细胞仪检测结果显示，Hpx组与模型组在12 h时细胞凋亡率无明显差异。Hpx组24 h凋亡细胞百分率较模型组明显降低（P < 0.01）。与模型组比较，Hpx组HO-1表达在24 h降低（P < 0.01），caspase-3表达在12和24 h均降低（P < 0.011和P < 0.05）。结论Hpx通过抗氧化及降低HO-1和caspase-3的表达，对血液暴露的神经细胞具有保护作用。
Background A growing body of experimental evidence suggests that hemin released from heme is a potent oxidant and accumulates in intracranial hematomas. Hemopexin (Hpx) decreases hemin accumulation and catabolism by nerve cells. In previous study, we observed that Hpx gene knockout aggravated striatal injury and worsened behavioral deficits of mice subjected to intracerebral hemorrhage. Aim To examine the effect of Hpx on oxidative damage and apoptosis in cultured nerve cells with blood clot. Methods Neuron and glial cells were transfected with adenoviral Hpx gene. Transfected primary neuron-glial cells were co-cultured with 50 μl of arterial blood clot using insert transwells. The sham group was co-coulture with 50 μl of DMEM/F12, which contained 28 μl of serum; the control group was transfected with adenoviral vector. At 12 and 24 h, the level of malonaldehyde (MDA), surperoxide dismutase (SOD) concentration, glutathione (GSH), apoptosis, expression of HO-1 and caspase-3 were detected. Results MDA level was decreased (P < 0.01) whereas SOD and GSH concentration were increased in the Hpx group (P < 0.05 and P < 0.01, respectively). Results of flow cytometry revealed no significant difference in apoptosis between the Hpx group and model group at 12 h. However, the percentage of cells undergoing apoptosis in the Hpx group was decreased at 24 h compared with the model group (P < 0.01). HO-1 expression decreased in the Hpx group at 24 h (P < 0.01) while caspase-3 expression decreased at both 12 and 24 h (P < 0.011 and P < 0.05, respectively) compared with the model group. Conclusion Hpx protected nerve cells exposed to blood from injury by anti-oxidation and a decrease in the expression of HO-1 and caspase-3.