天津医药

天津医药 ›› 2021, Vol. 49 ›› Issue (4): 371-377.doi: 10.11958/20202542

• 实验研究 • 上一篇    下一篇

Xyloketal B对发育期大鼠惊厥后脑损伤的保护作用及机制研究#br#

陈芬芳，胡擎鹏△   

  1. 南华大学附属第二医院儿科（邮编421001）
  • 收稿日期:2020-09-16 修回日期:2020-12-04 出版日期:2021-04-15 发布日期:2021-04-16
  • 通讯作者: 胡擎鹏 E-mail:pengpeng000923@163.com
  • 作者简介:陈芬芳（1982），女，硕士，主治医师，主要从事小儿神经病学研究。E-mail：64653352@qq.com
  • 基金资助:
    湖南省自然科学基金面上项目（2019JJ40263）

The protective effect and mechanism of Xyloketal B on seizure-induced brain damage in developing rats#br#

CHEN Fen-fang, HU Qing-peng△   

  1. Department of Pediatrics, the Second Hospital, University of South China, Hengyang 421001, China
  • Received:2020-09-16 Revised:2020-12-04 Published:2021-04-15 Online:2021-04-16
  • Contact: HU Qing-peng E-mail:pengpeng000923@163.com

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摘要： 目的　探讨Xyloketal B（Xyl-B）对发育期大鼠惊厥后脑损伤的保护作用及对氧化应激通路的调控。方法　40只20日龄SD大鼠随机分为Control组、海人酸（KA）组、KA+Xyl-B（25 mg/kg）组及KA+Xyl-B（50 mg/kg）组，通过腹腔注射KA制作大鼠惊厥模型，Control组注射生理盐水；KA+Xyl-B各组在KA诱导惊厥前2 h腹腔注射Xyl-B预治疗。惊厥后24 h处死大鼠，取海马组织，采用免疫荧光检测成熟神经元和活化星形胶质细胞，实时荧光定量PCR（qPCR）和Western blot分别检测Kelch样环氧氯丙烷相关蛋白-1（Keap1）、核因子E2相关因子2（Nrf2）、血红素氧合酶1（HO-1）、依赖还原型辅酶/Ⅱ醌氧化还原酶1（NQO1）、caspase3、Bcl-2和白细胞介素（IL）-6 mRNA及蛋白的表达。结果　与Control组比较，KA组神经元数量明显减少，活化星形胶质细胞显著增多；Keap1 mRNA和蛋白表达水平升高，Nrf2、HO-1、NQO1 mRNA和蛋白表达水平降低；caspase3和IL-6 mRNA和蛋白表达水平升高，Bcl-2 mRNA和蛋白表达水平降低（P＜0.05）。Xyl-B干预可逆转惊厥后上述病理变化，且KA+Xyl-B（50 mg/kg）组的干预效果整体上优于KA+Xyl-B（25 mg/kg）组。结论　Xyl-B能激活Nrf2抗氧化应激通路，抑制星形胶质细胞活化、炎性因子及凋亡蛋白的表达，对惊厥后脑损伤发挥保护作用。

关键词: 发作, 脑损伤, NF-E2相关因子2, 海马, 神经元, 星形细胞, Xyloketal B

Abstract: Objective　To explore the protective effect of marine natural product Xyloketal B (Xyl-B) on brain damage after seizure in developing rats and its regulation on oxidative stress pathway. Methods　Forty SD rats on the 20th day were randomly divided into 4 groups, control group, KA group, KA+Xyl-B (25 mg/kg) group and KA+Xyl-B (50 mg/kg) group. The rat seizure model was constructed through intraperitoneal injection of kainic acid. The control group was injected with saline only. In KA+Xyl-B group, Xyl-B was administered intraperitoneally 2 hours before KA induced seizure. Rats were sacrificed 24 hours after seizure, hippocampal tissue was separated. The number of mature neurons and activated astrocytes were detected by immunofluorescence. The mRNA and protein expression levels of Keap1, Nrf2, HO-1, NQO1, caspase3, Bcl-2 and IL6 were detected by qPCR and Western blot assay, respectively. Results　The immunofluorescence results showed that the number of neurons decreased significantly and the activated astrocytes increased significantly in the KA group compared with those of the control group. Xyloketal B treatment can increase the number of mature neurons and  the activated astrocytes. Compared with the control group, the mRNA and protein expression levels of Keap1 increased significantly, while the expression levels of Nrf2, HO-1 and NQO1 were significantly reduced. Xyloketal B treatment could reverse the mRNA and protein levels after KA-induced seizure. The mRNA and protein expression levels of caspase3 and IL-6 increased significantly, Bcl-2 decreased significantly in the KA group (P＜0.05). Xyloketal B treatment (50 mg/kg) can inhibit the expression levels of caspase3 and IL-6 and increase the expression of Bcl-2, which showed a better therapeutic effect than those of Xyl-B (25 mg/kg). Conclusion　Xyloketal B can activate the Nrf2 anti-oxidative stress pathway, inhibit the activation of astrocytes, inflammatory response and the expression of apoptosis protein, increase anti-apoptosis protein and surviving neurons, and thus play a protective role against brain damage after seizure.

Key words: seizures, brain injuries, NF-E2-related factor 2, hippocampus, neurons, astrocytes, Xyloketal B

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