积雪草酸对实验性蛛网膜下腔出血大鼠脑损伤的保护作用

doi:10.11958/20231956

天津医药 ›› 2024, Vol. 52 ›› Issue (3): 285-289.doi: 10.11958/20231956

积雪草酸对实验性蛛网膜下腔出血大鼠脑损伤的保护作用

武潇潇¹(), 胡玉鲲², 吴江³^,^△()

1.苏州大学苏州医学院（邮编215000）
2.南通大学附属常熟医院
3.苏州大学附属第一医院

收稿日期:2023-12-10 修回日期:2023-12-25 出版日期:2024-03-15 发布日期:2024-03-13
通讯作者: ^△E-mail：szjiangwu@163.com
作者简介:武潇潇（2001），女，本科在读，主要从事脑血管疾病及相关机制的研究。E-mail：2628962403@qq.com
基金资助:
国家自然科学基金资助项目(82171294);苏州大学“大学生创新创业训练计划”(202210285077Z)

Protective effect of asiatic acid on brain injury after experimental subarachnoidhemorrhage in rats

WU Xiaoxiao¹(), HU Yukun², WU Jiang³^,^△()

1. Suzhou Medical College of Soochow University, Suzhou 215000, China
2. the Affiliated Changshu Hospital of Nantong University
3. the First Affiliated Hospital of Soochow University

Received:2023-12-10 Revised:2023-12-25 Published:2024-03-15 Online:2024-03-13
Contact: ^△E-mail: szjiangwu@163.com

武潇潇, 胡玉鲲, 吴江. 积雪草酸对实验性蛛网膜下腔出血大鼠脑损伤的保护作用[J]. 天津医药, 2024, 52(3): 285-289.

WU Xiaoxiao, HU Yukun, WU Jiang. Protective effect of asiatic acid on brain injury after experimental subarachnoidhemorrhage in rats[J]. Tianjin Medical Journal, 2024, 52(3): 285-289.

摘要/Abstract

摘要：

目的探讨积雪草酸（AA）对实验性蛛网膜下腔出血（SAH）大鼠脑损伤的神经保护作用。方法将108只成年SD大鼠分为假手术（Sham）1组、SAH+Vehicle（空载）组与SAH+AA组，每组36只；42只大鼠分为Sham2组及SAH后3、6、12、24、48、72 h组，每组6只。除Sham组外，其余各组采用单侧颈外动脉线刺法建立SAH模型，造模后SAH+AA组灌胃AA溶液（30 mg/kg）。采用踏空实验和改良Garcia评分评估神经行为学改变，Western blot检测脑组织谷胱甘肽过氧化物酶4（GPX4）蛋白表达，酶联免疫吸附试验检测脑组织谷胱甘肽（GSH）和丙二醛（MDA）浓度，Fluoro-Jade B染色检测神经元死亡情况。结果与Sham1组相比，SAH+Vehicle组踏空步数比例明显增多且改良Garcia评分明显降低，脑组织GPX4蛋白表达水平和GSH含量降低，MDA含量升高，死亡神经元数目增多（均P<0.05）。与SAH+Vehicle组相比，SAH+AA组踏空步数比例下降且改良Garcia评分升高，脑组织GPX4蛋白表达升高，MDA含量下降，GSH含量升高，死亡神经元数目明显下降（P<0.05）。结论 AA可能通过抑制脂质过氧化减轻大鼠SAH后的脑损伤。

关键词: 积雪草酸, 脂质过氧化, 蛛网膜下腔出血, 脑损伤

Abstract:

Objective To explore the neuroprotective effect of asiatic acid (AA) on brain damage after experimental subarachnoid hemorrhage (SAH) in rats. Methods A total of 108 adult SD rats were divided into the sham1 group, the SAH+vehicle group and the SAH+AA group, with 36 rats in each group. The 42 rats were divided into the sham2 group, 3, 6, 12, 24, 48 and 72 h after SAH groups, with 6 rats in each group. Except the sham group, SAH model was established by unilateral external carotid artery puncture method in other groups. After modeling, the SAH+AA group was given AA solution (30 mg/kg) by gavage. Neurobehavioral changes were assessed by foot fault test and modified Garcia score. Western blot assay was used to detect the protein level of glutathione peroxidase 4 (GPX4) in brain tissue. ELISA was used to detect the concentrations of glutathione (GSH) and malondialdehyde (MDA). Fluoro Jade B (FJB) staining was used to detect the neuronal death. Results Compared with the sham1 group, the SAH+vehicle group showed a significant increase in the proportion of empty steps and a significant decrease in the modified Garcia score, a significant decrease in GPX4 protein levels, a significant increase in MDA concentration (P<0.05), a decrease in GSH concentration (P<0.01) and a significant increase in the number of dead neurons (P<0.05). Compared with the SAH+vehicle group, a significant decrease in the proportion of empty steps, a significant increase in the modified Garcia score, a significant increase in GPX4 protein level, a significant decrease in MDA concentration, a significant increase in GSH concentration (P<0.05) and a significant decrease in the number of dead neurons in the SAH+AA group (P<0.05). Conclusion AA may reduce brain injury after SAH in rats by inhibiting lipid peroxidation.

Key words: asiatic acid, lipid peroxidation, subarachnoid hemorrhage, brain damage

中图分类号:

R743.35

图/表 6

表1 各组大鼠不同时点踏空步数比例比较（%，$\bar{x} \pm s$）

Tab.1 Comparison of treadle proportion of rats between three groups

组别	n	造模前	SAH后3 d	SAH后5 d	SAH后7 d	SAH后14 d
Sham1组	12	1.58±0.90	1.50±1.45	1.25±0.97	0.75±0.75	0.67±0.89
SAH+Vehicle组	12	1.50±1.31	18.42±1.56^a	15.33±1.44^a	12.83±1.59^a	8.67±1.61^a
SAH+AA组	12	1.67±0.98	8.75±1.91^b	6.25±1.42^b	4.91±1.88^b	3.00±0.85^b
F		0.071	316.300^＊＊	365.900^＊＊	204.800^＊＊	147.900^＊＊

表2 各组大鼠改良Garcia评分比较（分，$\bar{x} \pm s$）

Tab.2 Comparison of improved Garcia scores between three groups of rats

组别	n	造模前	SAH后3 d	SAH后5 d	SAH后7 d
Sham1组	12	17.8±0.5	17.5±0.5	17.6±0.5	17.6±0.5
SAH+Vehicle组	12	17.5±0.5	12.3±1.4^a	13.3±1.6^a	14.1±1.6^a
SAH+AA组	12	17.6±0.5	14.9±0.7^b	15.8±0.7^b	16.6±0.8^b
F		0.786	90.460^＊＊	51.090^＊＊	35.020^＊＊

图1 SAH后不同时点大鼠脑组织中GPX4蛋白表达变化 A：Sham2组；B：SAH后3 h组；C：SAH后6 h组；D：SAH后12 h组；E：SAH后24 h组；F：SAH后48 h组；G：SAH后72 h组。a与Sham2组比较，P<0.05。

Fig.1 Changes of GPX4 protein expression in rat brain at different time points after SAH

图2 各组大鼠脑组织FJB染色结果（400×）

Fig.2 Results of FJB staining in brain tissue of rats in each group (400×)

图3 SAH后大鼠脑组织中脂质过氧化标志物的变化 A：Sham1组；B：SAH+Vehicle组；C：SAH+AA组。

Fig.3 The expression level of lipid peroxidation marker in brain tissue after SAH

表3 各组大鼠脑组织中脂质过氧化蛋白表达水平比较（$\bar{x} \pm s$）

Tab.3 Comparison of the expression level of lipid peroxidase in brain tissue between three groups of rats

组别	n	GPX4	MDA/（μmol/g）	GSH/（mg/g）
Sham1组	6	1.00±0.11	5.34±0.51	9.27±0.92
SAH+Vehicle组	6	0.57±0.06^a	10.77±1.53^a	6.93±1.06^a
SAH+AA组	6	0.71±0.09^b	7.10±0.46^b	8.26±0.92^b
F		37.870^＊＊	49.050^＊＊	10.260^＊＊

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积雪草酸对实验性蛛网膜下腔出血大鼠脑损伤的保护作用

Protective effect of asiatic acid on brain injury after experimental subarachnoidhemorrhage in rats

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