瑞马唑仑调控Nrf2/GPX4通路对脓毒性休克大鼠循环功能的影响

doi:10.11958/20241809

天津医药 ›› 2025, Vol. 53 ›› Issue (4): 349-354.doi: 10.11958/20241809

瑞马唑仑调控Nrf2/GPX4通路对脓毒性休克大鼠循环功能的影响

郝妍妍¹(), 张宇², 白耀武, 石东海¹^,^△()

1 唐山市妇幼保健院麻醉科（邮编063000）
2 唐山市妇幼保健院超声科（邮编063000）

收稿日期:2024-11-11 修回日期:2025-02-06 出版日期:2025-04-15 发布日期:2025-04-17
通讯作者: ^△ E-mail：xzztx71@163.com
作者简介:郝妍妍（1991），女，主治医师，主要从事临床麻醉、疼痛诊疗方面研究。E-mail：pzvicb@163.com
基金资助:
河北省医学科学研究重点课题计划项目(20241254)

Effects of remimazolam regulating the Nrf2/GPX4 pathway on circulatory function in septic shock rats

HAO Yanyan¹(), ZHANG Yu², BAI Yaowu, SHI Donghai¹^,^△()

1 Department of Anesthesiology, Tangshan Maternal and Child Health Hospital, Tangshan 063000, China
2 Department of Ultrasound, Tangshan Maternal and Child Health Hospital, Tangshan 063000, China

Received:2024-11-11 Revised:2025-02-06 Published:2025-04-15 Online:2025-04-17
Contact: ^△ E-mail：xzztx71@163.com

郝妍妍, 张宇, 白耀武, 石东海. 瑞马唑仑调控Nrf2/GPX4通路对脓毒性休克大鼠循环功能的影响[J]. 天津医药, 2025, 53(4): 349-354.

HAO Yanyan, ZHANG Yu, BAI Yaowu, SHI Donghai. Effects of remimazolam regulating the Nrf2/GPX4 pathway on circulatory function in septic shock rats[J]. Tianjin Medical Journal, 2025, 53(4): 349-354.

摘要/Abstract

摘要：

目的探讨瑞马唑仑对脓毒性休克大鼠循环功能的影响与机制。方法 72只SPF级大鼠随机分为对照组、模型组、地塞米松组、瑞马唑仑低剂量组、瑞马唑仑高剂量组、瑞马唑仑高剂量+Nrf2抑制剂（ML385）组，每组12只。采用股静脉滴注10 mg/kg的脂多糖诱导建立脓毒性休克大鼠模型。造模6 h后，检测大鼠平均动脉压（MAP）和心率（HR）；酶联免疫吸附试验测定血清乳酸（Lac）、肿瘤坏死因子-α（TNF-α）、白细胞介素（IL）-1β、IL-6、一氧化氮（NO）和内皮素1（ET-1）水平；苏木精-伊红（HE）染色观察血管组织形态学变化；TUNEL染色观察血管内皮细胞凋亡情况；二氢乙锭荧光探针检测血管组织活性氧（ROS）水平；比色法检测血管组织丙二醛（MDA）含量和超氧化物歧化酶（SOD）活性；Western blot检测血管组织核因子E2相关因子2（Nrf2）、谷胱甘肽过氧化物酶4（GPX4）蛋白表达。结果与对照组相比，模型组大鼠MAP、血管组织SOD活性、Nrf2、GPX4蛋白水平降低，HR和血清Lac、NO、ET-1、TNF-α、IL-1β、IL-6水平、血管内皮细胞凋亡率、血管组织ROS水平、MDA含量升高（P<0.05）；与模型组比较，地塞米松组，瑞马唑仑低、高剂量组大鼠MAP、血管组织SOD活性、Nrf2、GPX4蛋白水平升高，HR和血清Lac、NO、ET-1、TNF-α、IL-1β、IL-6水平、血管内皮细胞凋亡率、血管组织ROS水平、MDA含量降低（P<0.05）；Nrf2抑制剂ML385明显减弱瑞马唑仑对脓毒性休克大鼠的保护作用（P<0.05）。结论瑞马唑仑可能通过激活Nrf2/GPX4通路，抑制炎症反应和氧化应激，减轻血管内皮细胞损伤，进而改善脓毒性休克大鼠循环功能。

关键词: 休克, 脓毒性, 内皮细胞, 瑞马唑仑, 核因子E2相关因子2, 谷胱甘肽过氧化物酶4

Abstract:

Objective To investigate the effect and mechanism of remimazolam on circulatory function in septic shock rats. Methods Seventy-two SPF grade rats were randomly divided into the control group, the model group, the dexamethasone group, the low and high dose remimazolam groups and the high-dose remimazolam+Nrf2 inhibitor (ML385) group, with 12 rats in each group. The septic shock rat model was established by intravenous infusion of 10 mg/kg lipopolysaccharide (LPS). After 6 hours of modeling, the mean arterial pressure (MAP) and heart rate (HR) of rats were measured. Enzyme linked immunosorbent assay (ELISA) method was applied to measure serum levels of lactic acid (Lac), tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-6, nitric oxide (NO), and endothelin-1 (ET-1). Hematoxylin-eosin (HE) staining was applied to observe morphological changes in vascular tissue. TUNEL staining was applied to observe the apoptosis of vascular endothelial cells. DHE fluorescent probe was used to detect the level of ROS in vascular tissue. The colorimetric method was applied to detect the contents of MDA and the activity of SOD in vascular tissue. Western blot assay was applied to detect the protein expression of nuclear factor E2-related factor 2 (Nrf2) and glutathione peroxidase 4 (GPX4) in vascular tissue. Results Compared with the control group, MAP, SOD activity in vascular tissue, Nrf2 and GPX4 protein levels were lower in the model group, while HR, serum Lac, NO, ET-1, TNF-α, IL-1β, IL-6 levels, endothelial cell apoptosis rate, ROS level in vascular tissue and MDA content were higher (P<0.05). Compared with the model group, MAP, SOD activity in vascular tissue, Nrf2 and GPX4 protein levels were higher in the dexamethasone group and in the low and high dose remimazolam groups, while HR, serum Lac, NO, ET-1, TNF-α, IL-1β, IL-6 levels, endothelial cell apoptosis rate, ROS level in vascular tissue and MDA content were lower (P<0.05). Nrf2 inhibitor ML385 greatly reduced the protective effect of remimazolam on septic shock rats (P<0.05). Conclusion Remimazolam may improve circulatory function in septic shock rats by activating the Nrf2/GPX4 pathway, inhibiting inflammatory response and oxidative stress, reducing endothelial cell damage.

Key words: shock, septic, endothelial cells, remimazolam, nuclear factor E2 related factor 2, glutathione peroxidase 4

中图分类号:

R631

图/表 9

参考文献 26

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组别	MAP/mmHg	HR/（次/min）	Lac/（mmol/L）
对照组	98.17±2.41	317.50±12.65	1.86±0.31
模型组	47.00±3.16^a	490.42±18.43^a	4.90±0.57^a
地塞米松组	85.42±2.64^b	336.92±17.30^b	2.39±0.36^b
瑞马唑仑低剂量组	69.00±2.49^bc	444.58±13.28^bc	4.11±0.51^bc
瑞马唑仑高剂量组	83.67±4.08^bd	347.42±14.58^bd	2.52±0.40^bd
瑞马唑仑高剂量+ML385组	55.50±4.30^e	470.50±13.60^e	4.60±0.54^e
F	213.415^＊＊	150.423^＊＊	47.693^＊＊

组别	MAP/mmHg	HR/（次/min）	Lac/（mmol/L）
对照组	98.17±2.41	317.50±12.65	1.86±0.31
模型组	47.00±3.16^a	490.42±18.43^a	4.90±0.57^a
地塞米松组	85.42±2.64^b	336.92±17.30^b	2.39±0.36^b
瑞马唑仑低剂量组	69.00±2.49^bc	444.58±13.28^bc	4.11±0.51^bc
瑞马唑仑高剂量组	83.67±4.08^bd	347.42±14.58^bd	2.52±0.40^bd
瑞马唑仑高剂量+ML385组	55.50±4.30^e	470.50±13.60^e	4.60±0.54^e
F	213.415^＊＊	150.423^＊＊	47.693^＊＊

组别	NO/（μmol/L）	ET-1/（ng/L）
对照组	14.27±2.31	5.52±1.03
模型组	56.30±4.24^a	18.44±1.99^a
地塞米松组	24.15±3.08^b	8.39±1.17^b
瑞马唑仑低剂量组	39.42±3.76^bc	13.69±1.52^bc
瑞马唑仑高剂量组	26.93±3.25^bd	9.02±1.31^bd
瑞马唑仑高剂量+ML385组	48.60±4.10^e	15.74±1.68^e
F	123.114^＊＊	66.340^＊＊

组别	NO/（μmol/L）	ET-1/（ng/L）
对照组	14.27±2.31	5.52±1.03
模型组	56.30±4.24^a	18.44±1.99^a
地塞米松组	24.15±3.08^b	8.39±1.17^b
瑞马唑仑低剂量组	39.42±3.76^bc	13.69±1.52^bc
瑞马唑仑高剂量组	26.93±3.25^bd	9.02±1.31^bd
瑞马唑仑高剂量+ML385组	48.60±4.10^e	15.74±1.68^e
F	123.114^＊＊	66.340^＊＊

组别	TNF-α/（ng/L）	IL-1β/（ng/L）	IL-6/（ng/L）
对照组	45.60±5.72	22.48±4.19	27.75±5.21
模型组	149.44±9.83^a	97.30±8.06^a	126.49±10.30^a
地塞米松组	66.32±6.40^b	38.65±5.25^b	44.38±6.45^b
瑞马唑仑低剂量组	101.25±9.29^bc	77.24±7.13^bc	89.20±9.10^bc
瑞马唑仑高剂量组	70.41±7.35^bd	42.18±6.22^bd	50.69±7.28^bd
瑞马唑仑高剂量+ML385组	120.23±9.64^e	85.67±7.90^e	107.83±9.92^e
F	133.122^＊＊	124.447^＊＊	135.321^＊＊

瑞马唑仑调控Nrf2/GPX4通路对脓毒性休克大鼠循环功能的影响

Effects of remimazolam regulating the Nrf2/GPX4 pathway on circulatory function in septic shock rats

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组别	血管内皮细胞凋亡率/%		ROS/ （平均荧光强度）
对照组	3.40±0.62		55.74±6.82
模型组	17.82±1.13^a		190.60±10.34^a
地塞米松组	6.95±0.80^b		84.95±8.15^b
瑞马唑仑低剂量组	12.04±0.91^bc		136.27±9.61^bc
瑞马唑仑高剂量组	7.32±0.85^bd		90.18±8.32^bd
瑞马唑仑高剂量+ML385组	14.11±1.02^e		165.35±10.17^e
F	208.706^＊＊		201.233^＊＊
组别		MDA/ （nmol/mg）		SOD/ （U/mg）
对照组		5.95±0.80		38.60±4.15
模型组		14.46±1.52^a		12.75±2.30^a
地塞米松组		7.18±0.93^b		30.22±3.26^b
瑞马唑仑低剂量组		10.69±1.26^bc		18.18±2.79^bc
瑞马唑仑高剂量组		7.55±1.01^bd		28.95±3.10^bd
瑞马唑仑高剂量+ML385组		11.89±1.33^e		15.40±2.84^e
F		46.962^＊＊		62.754^＊＊

组别	Nrf2/Lamin B1	GPX4/GAPDH
对照组	0.57±0.06	0.73±0.08
模型组	0.20±0.03^a	0.25±0.04^a
地塞米松组	0.45±0.06^b	0.60±0.07^b
瑞马唑仑低剂量组	0.31±0.04^bc	0.39±0.05^bc
瑞马唑仑高剂量组	0.42±0.05^bd	0.56±0.06^bd
瑞马唑仑高剂量+ML385组	0.25±0.04^e	0.32±0.05^e
F	99.902^＊＊	113.706^＊＊

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