何银,刘春雨,何迟迟,赵锦,孙艳辉,徐红帅,蔡希强,李怡芳,栗原博,何蓉蓉
HE Yin,LIU Chun-yu,HE Chi-chi,ZHAO Jin,SUN Yan-hui,XU Hong-shuai,CAI Xi-qiang,LI Yi-fang,KURIHARA Hiroshi,HE Rong-rong

• 1:遂川县人民医院
• 2:暨南大学广东省疾病易感性及中医药研发工程技术研究中心
• 3:解放军第一六三医院
• 4:锦州市中心医院

摘要(Abstract)：

该研究制备了对乙酰氨基酚(APAP)诱导小鼠药物性肝损伤(DILI)模型,探讨了扶正养肝合剂对DILI的保护作用和机制。小鼠随机分为空白组,模型组,阳性药组,扶正养肝合剂高、低剂量组(0.24,0.12 g·kg~(-1))。小鼠连续灌胃给药21 d,最后一次给药16 h后,一次性腹腔注射APAP(600 mg·kg~(-1)),建立急性DILI小鼠模型,观察扶正养肝合剂对DILI小鼠肝脏质量和肝重比的影响;考察血清中谷丙转氨酶(ALT)和谷草转氨酶(AST)含量以及肝组织中丙二醛(MDA)、谷胱甘肽(GSH)及还原型辅酶Ⅱ(NADPH)水平;用Q-PCR法测定肝组织中ptgs2 mRNA水平;用Western blot法检测肝脏中SLC7A11和GPX4蛋白水平;并用HE染色、免疫组化、TUNEL法检测肝脏切片病理学变化。结果表明,扶正养肝合剂能够缓解APAP诱导的小鼠肝脏肿大及肝重比增加,降低DILI小鼠血清中ALT和AST含量,同时抑制MDA产生,升高NADPH及GSH水平,下调ptgs2 mRNA水平,上调SLC7A11和GPX4蛋白水平。结果表明,扶正养肝合剂通过抑制ferroptosis发挥对DILI的保护作用。
The model of drug-induced liver injury(DILI) induced by acetaminophen(APAP) in mice was established to investigate the anti-oxidation and anti-ferroptosis mechanisms of Fuzheng Yanggan Mixture on DILI. C57 BL/6 mice were randomly divided into five groups: control group, model group, positive group, and low and high-dose Fuzheng Yanggan Mixture groups(0.12, 0.24 g·kg~(-1)). Mice were intragastrically administration with Fuzheng Yanggan Mixture(0.12, 0.24 g·kg~(-1)) once per day for 21 consecutive days, and at the same time, mice were weighted every day. The mice were injected intraperitoneally with 600 mg·kg~(-1) of APAP to establish a mouse model of acute DILI after 16 h from the last administration of Fuzheng Yanggan Mixture. After 6 h from APAP challenge, the experimental animals were weighted and sacrificed to collect blood and liver tissue samples. And then, the effect of Fuzheng Yanggan Mixture on liver weight and the liver weight ratio of mice were examined; the content of alanine aminotransferase(ALT) and aspartate aminotransferase(AST) in the serum and the level of malondialdehyde(MDA), glutathione(GSH) and nicotinamide adenine dinucleotide phosphate(NADPH) in the liver tissue were measured. Prostaglandinendoperoxide synthase 2(ptgs2) mRNA level in liver tissues was detected by Q-PCR,and protein expression levels of SLC7 A11 and GPX4 in liver tissues were detected by Western blot. Moreover, HE staining, immunohistochemical assay and TUNEL staining were used to observe pathological changes of the liver tissue sections. It is found that Fuzheng Yanggan Mixture could relieve APAP-induced liver enlargement and inhibit hepatic weight ratio increase. Compared with model group, the mice in Fuzheng Yanggan Mixture groups showed decreases in the content of ALT, AST and MDA, and increases in the content of GSH and NADPH. What is more, Fuzheng Yanggan Mixture could down-regulate ptgs2 mRNA level and up-regulate SLC7 A11 and GPX4 protein levels. All of the results lead to a conclusion that Fuzheng Yanggan Mixture plays a protective effect on DILI in mice, which may be associated with the inhibition of ferroptosis.

关键词(KeyWords)： 扶正养肝合剂;对乙酰氨基酚(APAP);肝损伤;氧化应激;ferroptosis
Fuzheng Yanggan Mixture;acetaminophen(APAP);liver injury;oxidative stress;ferroptosis

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(81622050,81573675,81873209)

作者(Author): 何银,刘春雨,何迟迟,赵锦,孙艳辉,徐红帅,蔡希强,李怡芳,栗原博,何蓉蓉
HE Yin,LIU Chun-yu,HE Chi-chi,ZHAO Jin,SUN Yan-hui,XU Hong-shuai,CAI Xi-qiang,LI Yi-fang,KURIHARA Hiroshi,HE Rong-rong

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