Nrf2-GPX4介导的铁死亡通路参与右美托咪定对脑出血大鼠神经保护作用的机制研究

doi:10.11958/20212825

天津医药 ›› 2022, Vol. 50 ›› Issue (8): 817-821.doi: 10.11958/20212825

Nrf2-GPX4介导的铁死亡通路参与右美托咪定对脑出血大鼠神经保护作用的机制研究

李秋畅(), 闫顺昌, 蒙亚珍, 顾云霞, 宁巧明, 李娜, 王志华()

海南省人民医院麻醉科（邮编570311）

收稿日期:2021-12-23 修回日期:2022-03-28 出版日期:2022-08-15 发布日期:2022-08-12
通讯作者: 王志华 E-mail:liqiuchang814@163.com;3235885642@qq.com
作者简介:李秋畅（1987）,男,主治医师,主要从事超声引导神经阻滞方面研究。E-mail： liqiuchang814@163.com
基金资助:
2019年海南省基础与应用基础研究计划(2019RC365)

Neuroprotective effects of dexmedetomidine on intracerebral hemorrhage of rats by Nrf2-GPX4 mediated iron death pathway

LI Qiuchang(), YAN Shunchang, MENG Yazhen, GU Yunxia, NING Qiaoming, LI Na, WANG Zhihua()

Department of Anesthesiology, Hainan Provincial People's Hospital, Haikou 570311, China

Received:2021-12-23 Revised:2022-03-28 Published:2022-08-15 Online:2022-08-12
Contact: WANG Zhihua E-mail:liqiuchang814@163.com;3235885642@qq.com

李秋畅, 闫顺昌, 蒙亚珍, 顾云霞, 宁巧明, 李娜, 王志华. Nrf2-GPX4介导的铁死亡通路参与右美托咪定对脑出血大鼠神经保护作用的机制研究[J]. 天津医药, 2022, 50(8): 817-821.

LI Qiuchang, YAN Shunchang, MENG Yazhen, GU Yunxia, NING Qiaoming, LI Na, WANG Zhihua. Neuroprotective effects of dexmedetomidine on intracerebral hemorrhage of rats by Nrf2-GPX4 mediated iron death pathway[J]. Tianjin Medical Journal, 2022, 50(8): 817-821.

摘要/Abstract

摘要：

目的探究核因子E2相关因子2/谷胱甘肽过氧化物酶4（Nrf2-GPX4）介导的铁死亡通路参与右美托咪定（Dex）对脑出血（ICH）大鼠发挥神经保护作用的机制。方法将100只SD大鼠按随机数字表法分为Sham组、ICH组（模型组）、Dex-L组（Dex 50 μg/kg）、Dex-H组（Dex 100 μg/kg）、Dex-H+ML385组（Nrf2抑制剂ML385,30 mg/kg）,每组20只。除Sham组外,其余组通过自体血注射法建立ICH模型;Dex-L组、Dex-H组及Dex-H+ML385组于术前30 min腹腔注射相应Dex或ML385,Sham组和ICH组则注射等量的生理盐水。术后2 h Zea Longa评分评定大鼠神经功能损伤。试剂盒检测血肿周围脑组织谷胱甘肽（GSH）、丙二醛（MDA）、铁离子含量;称质量检测大鼠血肿周围脑含水量;HE染色、Nissl染色、普鲁士蓝染色分别观察血肿周围脑组织病理学、神经细胞损伤及铁沉积情况;Western blot检测脑组织GPX4、胱氨酸/谷氨酸逆向转运蛋白（xCT）、Nrf2表达。结果相较于Sham组,神经功能缺损评分、MDA、铁离子含量、脑含水量、脑组织病理损伤、铁沉积在ICH组明显增加,GSH含量、神经细胞数、GPX4、xCT、Nrf2表达水平明显减少（均P<0.05）;相较于ICH组,神经功能缺损评分、MDA、铁离子含量、脑含水量、脑组织病理损伤、铁沉积在Dex-L组与Dex-H组依次明显降低,GSH含量、神经细胞数、GPX4、xCT、Nrf2表达水平明显增加（均P<0.05）;ML385可逆转Dex-H对神经功能、铁沉积、脑损伤的改善。结论 Dex通过激活Nrf2-GPX4通路来抑制铁死亡,从而对ICH大鼠发挥神经保护的作用。

Abstract:

Objective To explore the mechanism of Nrf2-GPX4-mediated iron death pathway in the neuroprotective effect of dexmedetomidine (Dex) on intracerebral hemorrhage (ICH) in rats. Methods A total of 100 rats were randomly divided into the sham group, the ICH group (model group), the Dex-L group (Dex 50 μg/kg), the Dex-H group (Dex 100 μg/kg) and the Dex-H+ML385 group (Nrf2 inhibitor ML385, 30 mg/kg), with 20 rats in each group. Except for the sham group, ICH model was established by autologous blood injection in the other groups. In the Dex-L group, the Dex-H group and the Dex-H+ML385 group, the corresponding Dex or ML385 was injected intraperitoneally 30 minutes before operation, and the sham group and the ICH group were injected with the same amount of normal saline. Zea Longa 5 scoring method was used to evaluate rat nerve function damage. The contents of glutathione (GSH), malondialdehyde (MDA) and iron ions in brain tissue around the hematoma were detected by the kit. The brain water content around the hematoma was measured. HE staining, Nissl staining and Prussian blue staining were used to observe the pathology, nerve cell damage and iron deposition around the hematoma. Western blot assay was used to detect the expression levels of GPX4, cystine/glutamate antiporter system light chain (xCT) and Nrf2 in brain tissue. Results Compared with the sham group, the neurological deficit score, MDA, iron content, brain water content, brain tissue pathological damage and iron deposition were significantly increased in the ICH group, while the GSH content, nerve cell number, GPX4, xCT and Nrf2 expression levels were significantly decreased in the ICH group (P<0.05). Compared with the ICH group, the neurological deficit score, MDA, iron content, brain water content, brain tissue pathological damage and iron deposition were significantly reduced in turn in the Dex-L group and the Dex-H group, and the GSH content, nerve cell number, GPX4, xCT and Nrf2 expression levels were significantly increased (P<0.05). ML385 reversed improvements in neurological function, iron deposition and brain damage caused by Dex-H. Conclusion Dexmedetomidine inhibits iron death by activating the Nrf2-GPX4 pathway, thereby exerting a neuroprotective effect on ICH rats.

Key words: NF-E2-related factor 2, cerebral hemorrhage, glutathione peroxidase, dexmedetomidine, iron death

中图分类号:

R329.21

图/表 6

表1 各组大鼠脑组织铁离子、MDA、GSH含量比较（n=5,$\bar{x}±s$）

Tab.1 Comparison of iron ion, MDA and GSH content in brain tissue of rats between the five groups

组别	MDA（µmol/g）	铁离子（μg/g）	GSH（mg/g）
Sham组	5.01±0.65	40.28±3.22	3.49±0.33
ICH组	16.95±1.03^a	93.50±4.29^a	0.84±0.26^a
Dex-L组	10.73±0.82^b	76.84±4.03^b	1.92±0.30^b
Dex-H组	7.12±0.68^bc	58.08±3.69^bc	2.80±0.35^bc
Dex-H+ML385组	12.87±1.10^d	81.23±4.22^d	1.12±0.28^d
F	145.026^＊＊	143.132^＊＊	66.601^＊＊

图1 各组大鼠脑组织病理情况变化（HE染色,×400）

Fig.1 Pathological changes of brain tissue in each group (HE staining, ×400)

图2 各组大鼠脑组织神经细胞损伤情况比较（Nissl染色,×400）

Fig.2 Comparison of nerve cell damage in brain tissue of rats between the five groups (Nissl staining, ×400)

图3 各组大鼠脑组织铁沉积比较（普鲁士蓝染色,×400）

Fig.3 Comparison of iron deposition in brain tissue of rats between the five groups (Prussian blue staining, ×400)

表2 各组大鼠脑组织GPX4、xCT、Nrf2表达水平比较（n=5,$\bar{x}±s$）

Tab.2 Comparison of the expression levels of GPX4, xCT and Nrf2 in brain tissue of rats between the five groups

组别	GPX4/β-actin	xCT/β-actin	Nrf2/β-actin
Sham组	1.67±0.25	1.08±0.10	1.40±0.21
ICH组	0.44±0.07^a	0.25±0.06^a	0.61±0.08^a
Dex-L组	0.83±0.11^b	0.56±0.07^b	0.94±0.10^b
Dex-H组	1.32±0.19^bc	0.83±0.09^bc	1.23±0.14^bc
Dex-H+ML385组	0.70±0.09^d	0.45±0.08^d	0.82±0.09^d
F	49.691^＊＊	80.326^＊＊	28.416^＊＊

图4 各组大鼠脑组织GPX4、xCT、Nrf2蛋白表达水平比较 A：Sham组;B：ICH组;C：Dex-L组;D：Dex-H组;E：Dex-H+ML385组。

Fig.4 Comparison of the expression levels of GPX4, xCT and Nrf2 in brain tissue of rats between the five groups

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Nrf2-GPX4介导的铁死亡通路参与右美托咪定对脑出血大鼠神经保护作用的机制研究

Neuroprotective effects of dexmedetomidine on intracerebral hemorrhage of rats by Nrf2-GPX4 mediated iron death pathway

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