1. 福建中医药大学中西医结合研究院,福州 350122;2. 福建省中西医结合老年性疾病重点实验室,福州 350122;3. 福建省高校中西医结合基础重点实验室,福州 350122
LIN Jiumao
1. Academy of Integrative Medicine, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, China;2. Fujian Key Laboratory of Integrative Medicine on Geriatrics, Fuzhou 350122, China; 3. Fujian Key Laboratory of Integrative Medicine Periodial Office, Fuzhou 350122, China1. Academy of Integrative Medicine, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, China;2. Fujian Key Laboratory of Integrative Medicine on Geriatrics, Fuzhou 350122, China; 3. Fujian Key Laboratory of Integrative Medicine Periodial Office, Fuzhou 350122, China
目的 探讨八宝丹(Ba Bao Dan,BBD)对阿霉素诱导的斑马鱼心脏损伤的作用。 方法 建立化疗药物阿霉素(doxorubicin,Dox)诱导的斑马鱼心肌损伤模型,在体视显微镜下观察 BBD 在不同浓度下对心包水肿比例及心率的干预作用;采用转基因斑马鱼 Tg(mpx:EGFP)原位观察 BBD 对心脏中性粒细胞浸润的抑制作用;观察BBD 对超氧化物歧化酶(superoxide dismutase,SOD)、过氧化氢酶( catalase,CAT)及丙二醛(malondialdehyde,MDA)的影响;Real-time qPCR 检测铁死亡相关因子包括谷胱甘肽过氧化物酶 4( glutathione peroxidase 4a,gpx4a)、前列腺素内过 氧 化 物 合 成 酶 ( prostaglandin-endoperoxide synthase 2, ptgs2 )、 花 生 四 烯 酸 5 脂 氧 合 酶 ( arachidonate5-lipoxygenase,alox5a)、酰基辅酶 A 合成酶长链家族成员 4(acyl-CoA synthetase long-chain family member 4,acsl4)的mRNA 表达;采用亚铁离子荧光探针检测斑马鱼心脏亚铁离子的积累情况。 结果 BBD 能够减缓 Dox 导致的斑马鱼心包水肿和心率减慢,减少心脏中性粒细胞的浸润(P<0. 05);降低 MDA 的浓度(P<0. 05),同时增强 SOD 和CAT 的活性(P<0. 05);显著抑制铁死亡,减少斑马鱼心脏亚铁离子的积累,抑制 ptgs2、alox5a、acsl4 的表达(P<0. 05),同时促进 gpx4a 的表达(P<0. 05)。 结论 BBD 通过抑制脂质过氧化的发生调节铁死亡从而减弱 Dox 诱导的斑马鱼心肌损伤,改善心功能。
Objective To investigate the effect of Ba Bao Dan(BBD) on cardiac injury induced by doxorubicin in zebrafish. Methods We induced a zebrafish myocardial injury model using the chemotherapeutic drug, doxorubicin. We then examined the effects of different concentrations of BBD on pericardial edema and heart rate under an in vivo microscope. We also examined the inhibitory effects of BBD on neutrophil infiltration in the heart in Tg ( mpx:EGFP) transgenic zebrafish. The impacts of BBD on superoxide dismutase, catalase, and malondialdehyde were observed. mRNA expression levels of ferroptosis-related factors, including glutathione peroxidase 4a ( gpx4a), prostaglandin-endoperoxide synthase 2 (ptgs2), arachidonate 5-lipoxygenase (alox5a), and acyl-CoA synthetase long-chain family member 4 (acsl4) were determined by Real-time quantitative polymerase chain reaction. The accumulation of ferrous ions in zebrafish heart was assessed using a fluorescent probe for ferrous ions. Results BBD alleviated doxorubicin-induced pericardial edema and bradycardia in zebrafish, reduced neutrophil infiltration in the heart ( P<0. 05 ), decreased malondialdehyde concentration (P<0. 05), and enhanced the activities of superoxide dismutase and catalase ( P<0. 05). BBD also significantly inhibited ferroptosis, reduced the accumulation of ferrous ions in the zebrafish heart, suppressed the expression of ptgs2, alox5a, and acsl4 (P<0. 05), and promoted the expression of gpx4a (P<0. 05). Conclusions BBD can attenuate doxorubicin-induced zebrafish myocardial injury and improve cardiac function by inhibiting lipid peroxidation and regulating ferroptosis.
李宪美,曾来凤,黄彬,余岚心,林久茂.基于斑马鱼模型探讨八宝丹对阿霉素所致心肌损伤的保护作用及机制[J].中国实验动物学报,2024,32(5):557~565.
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