补阳还五汤增强GPX4表达抑制铁死亡减轻脑缺血再灌注损伤

Study on the Effect of Buyang Huanwu Decoction on Enhancing the Expression of GPX4 and Inhibiting Ferroptosis to Alleviate Cerebral Ischemia-Reperfusion Injury

摘要

目的 探讨补阳还五汤减轻脑缺血再灌注损伤是否通过增强GPX4的表达抑制铁死亡的机制。方法 选取健康雄性SD大鼠48只,随机分为4组：假手术组（Sham组）、模型组（MCAO/R组）、补阳还五汤组（BYHWD组）、阳性对照药组（DFO组）。所有实验动物均建MCAO/R模型（假手术组仅剥离）。BYHWD组给予补阳还五汤灌胃处理,DFO组给予DFO腹腔注射处理,其余组予以生理盐水灌胃处理。再灌注72 h进行指标评定及取材。神经功能评分检测神经功能;TTC染色测定脑梗死面积;尼氏染色观察脑组织形态结构;生化试剂检测血清GSH、丙二醛（MDA）、脑组织铁含量;免疫荧光和Western blot检测谷胱甘肽过氧化物酶4（GPX4）的蛋白表达。结果 与假手术组相比,模型组大鼠神经功能损伤明显,脑梗死体积增多,脑组织病理学损伤明显,尼氏体数量减少,脑组织铁含量、血清MDA含量明显增加,血清GSH含量、脑组织GPX4的蛋白表达显著降低（P<0.05）;与模型组相比,补阳还五汤组大鼠神经功能明显好转,脑梗死体积降低,脑组织病理学损伤减轻,脑组织铁含量、血清MDA含量降低,血清GSH含量,脑组织GPX4的蛋白表达显著增加（P<0.05）。结论 补阳还五汤抑制脑缺血再灌注损伤可能通过增强GPX4的表达抑制铁死亡而发挥作用。

Abstract

Objective To explore the mechanism of Buyang Huanwu Decoction (BYHWD) in alleviating cerebral ischemia reperfusion injury by enhancing the expression of GPX4 and inhibiting iron death. Methods Fourty-eight healthy male SD rats were randomly divided into 4 groups: sham operation group (Sham), model group (MCAO/R), Buyang Huanwu decoction group (BYHWD), and positive control drug group (deferoxamine mesylate, DFO). All experimental animals were established MCAO/R model (sham operation group only stripped). Buyang Huanwu Decoction group (BYHWD) was given Buyang Huanwu Decoction by gavage, DFO group was given DFO by intraperitoneal injection, and other groups were given normal saline by gavage. The indexes were evaluated and materials were taken after 72 hours of reperfusion. Nerve function score was used to detect nerve function; the area of cerebral infarction was measured by TTC staining; Nessler's staining was used to observe the morphological structure of brain tissue; serum GSH, malondialdehyde (MDA) and iron content in brain tissue were detected with biochemical reagents; immunofluorescence and Western blot were used to detect the protein expression of glutathione peroxidase 4 (GPX4). Results Compared with the sham-operated group, the rats in the model group had obvious neurological damage, increased cerebral infarction volume, obvious brain histopathological damage, decreased the number of nissl bodies, increased brain iron content and serum MDA content, and significantly decreased serum GSH content and GPX4 protein expression in brain tissue (P<0.05); compared with the model group, the nerve function of the rats in the Buyang Huanwu Decoction group was significantly improved, the volume of cerebral infarction was reduced, the pathological damage of brain tissue was reduced, the content of iron in brain tissue and serum MDA were decreased, and the content of serum GSH and the protein expression of GPX4 in brain tissue were significantly increased (P<0.05). Conclusion Buyang Huanwu Decoction can inhibit cerebral ischemia reperfusion injury by increasing the expression of GPX4 and inhibiting ferroptosis.

关键词

脑缺血再灌注损伤 / 补阳还五汤 / 谷胱甘肽过氧化物酶4

Key words

cerebral ischemia reperfusion injury / Buyang Huanwu Decoction / glutathione peroxidase 4

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基金资助

中央引导地方科技发展资金项目（206Z7706G）; 河北省科技研发平台与新型研发机构建设专项（20567626H）