氧和血红蛋白诱导的神经元毒性及机制

betway必威登陆网址（betway.com ）学报››2021,Vol. 42››Issue (12): 889-893.DOI:10.3969/j.issn.2097-0005.2021.12.003

氧和血红蛋白诱导的神经元毒性及机制

陈琪¹, 陈鑫¹, 柴建薇¹, 张伟¹, 黄煜¹, 付晓艳²

^1.betway必威登陆网址（betway.com ）公共卫生学院，山东泰安　271016
^2.betway必威登陆网址（betway.com ）基础医学院，山东泰安 271000

收稿日期:2021-07-18出版日期:2021-12-25发布日期:2022-01-13
通讯作者:付晓艳
作者简介:陈琪，本科在读。
基金资助:
山东省大学生创新创业训练计划(S202010439076)

Mechanism of oxyhemoglobin-induced neuronal toxicity

Qi Chen¹, Xin Chen¹, Jianwei Chai¹, Wei Zhang¹, Yu Huang¹, Xiaoyan Fu²

^1.College of Basic Medicine，Shandong First Medical University ＆ Shandong Academy of Medical Sciences，Taian 271016，China
^2.College of Basic Medicine，Shandong First Medical University ＆ Shandong Academy of Medical Sciences，Taian 271000，China

Received:2021-07-18Online:2021-12-25Published:2022-01-13
Contact:Xiaoyan Fu

摘要/Abstract

摘要： 目的

评价氧合血红蛋白（oxyhemoglobin， Oxy-Hb）诱导神经元毒性的机制。

方法

体外构建Oxy-Hb神经元损伤模型，MTT法检测神经元活性；荧光探针检测自由基产生；western blotting检测蛋白表达。

结果

Oxy-Hb可剂量依赖性的抑制神经元活性，造成神经元形貌的改变；Oxy-Hb体外可时间和剂量依赖性的诱导自由基的产生和线粒体膜电位的耗散，进而导致DNA损伤和神经元凋亡，表现为caspase-3的活化和histone的磷酸化激活。

结论

Oxy-Hb可通过诱导自由基产生和线粒体功能紊乱启动神经元凋亡，对神经元有潜在的毒性，可能是脑出血造成脑损伤的重要机制之一。

关键词:氧合血红蛋白,脑出血,神经元毒性,神经元凋亡

Abstract: Objective

To evaluate oxyhemoglobin （Oxy-Hb）-induced neuronal toxicity and mechanism.

Methods

Oxy-Hb-induced neuronal damage model was established. MTT assay was used to examine neural viability， and Probe was used to examine the free radical. Western blotting was employed to detect protein expression.

Results

Oxy-Hb dose-dependently inhibited neural viability and changed the neural morphology. Oxy-Hb treatment time- and dose-dependently induced the generation of free radical and loss of mitochondrial membrane potential （MMP）， which subsequently caused DNA damage and neural apoptosis， as demonstrated by the caspase-3 activation and histone phosphorylation.

Conclusion

Oxy-Hb can induce significantly neural apoptosis by triggering free radical generation and MMP loss. These results indicate that Oxy-Hb shows potential neural toxicity， which may be one of the important mechanisms of cerebral hemorrhage injury.

Key words:oxyhemoglobin,cerebral hemorrhage,neural toxicity,neural apoptosis

中图分类号:

陈琪, 陈鑫, 柴建薇, 张伟, 黄煜, 付晓艳. 氧和血红蛋白诱导的神经元毒性及机制[J]. betway必威登陆网址（betway.com ）学报, 2021, 42(12): 889-893.

Qi Chen, Xin Chen, Jianwei Chai, Wei Zhang, Yu Huang, Xiaoyan Fu. Mechanism of oxyhemoglobin-induced neuronal toxicity[J]. Journal of Shandong First Medical Unversity & Shandong Academy of Medical Sciences, 2021, 42(12): 889-893.

图/表5

图1Oxy-Hb剂量依赖性的抑制神经元的活性（“*”表示P < 0.05 VS正常对照组，“**”表示P < 0.01 VS正常对照组）

图2Oxy-Hb对神经元形貌的影响（Tubulin抗体标记，免疫荧光染色观察神经元形貌的变化）

图3Oxy-Hb时间依赖性诱导了自由基的产生（绿色荧光：神经元实时ROS；红色荧光：超氧阴离子）

图4Oxy-Hb时间依赖性降低了线粒体膜电位（MMP）（红色荧光向绿色荧光的转变可反映MMP的变化）

图5Oxy-Hb时间依赖性诱导了caspas-3的激活和组蛋白的磷酸化（Western blotting）

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