大黄素干预PI3K/Akt/mTOR信号通路影响肝癌前病变大鼠铁死亡的作用机制  

Mechanism of Emodin on Ferroptosis by Interfering PI3K/Akt/mTOR Signaling Pathway in Rats with Precancerous Lesions of Liver Cancer

作　　者：陈凤菊[1] 鲜佳 南立静 常少雄 霍俊杰[1] 李继凯 CHEN Fengju;XIAN Jia;NAN Lijing;CHANG Shaoxiong;HUO Junjie;LI Jikai(Department of Radiology cind Chemotherapy,Second Af filiated Hospital of Xingtai Medical College,Xingtai Hebei 054000,China)

机构地区：[1]邢台医学高等专科学校第二附属医院放化疗科,河北邢台054000 [2]华北医疗健康集团邢台总医院脊柱骨科

出　　处：《联勤军事医学》2024年第5期361-366,375,共7页Military Medicine of Joint Logistics

基　　金：邢台市重点研发计划自筹项目(2023ZC127)。

摘　　要：目的 探究大黄素对二乙基亚硝胺(diethylnitrosamine, DEN)诱导的肝癌前病变大鼠磷脂酰肌醇3-激酶(phosphoinositide 3-kinase, PI3K)/蛋白激酶B(protein kinase B, Akt)/哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin, mTOR)信号转导通路的调控作用及对铁死亡的影响。方法 采用DEN诱发大鼠肝癌前病变进行肝癌前病变造模。随机将50只雄性SD大鼠分为对照组[未造模+10ml/(kg·d)生理盐水灌胃)]、DEN模型组[肝癌前病变造模+10 ml/(kg·d)生理盐水灌胃]、DEN+大黄素组[肝癌前病变造模+80 mg/(kg·d)大黄素浓缩液灌胃]、DEN+护肝片组[肝癌前病变造模+900 mg/kg护肝片灌服]和DEN+大黄素+护肝片组[肝癌前病变造模+80 mg/(kg·d)大黄素浓缩液灌胃+900 mg/kg护肝片灌服],每组各10只大鼠。各组大鼠连续干预治疗12周。生化分析法检测各组大鼠血清丙氨酸氨基转移酶(alanine aminotransferase, ALT)、天门冬氨酸氨基转移酶(aspartate aminotransferase, AST)、白蛋白(albumin, ALB)水平;比较各组大鼠的肝脏指数。酶联免疫吸附试验(enzyme-linked immunosorbent assay, ELISA)法检测各组大鼠血清白细胞介素1β(interleukin 1 beta, IL-1β)、肿瘤坏死因子α(tumor necrosis factor alpha, TNF-α)、IL-10含量。苏木精-伊红(hematoxylin-eosin, HE)染色法观察各组大鼠肝组织病理形态学改变;脱氧核糖核苷酸末端转移酶介导的缺口末端标记法(terminal deoxynucleotidyl transferase mediated dUTP nick-end labeling, TUNEL)分析各组大鼠肝组织细胞凋亡情况。免疫组织化学法检测各组大鼠肝组织中铁死亡中心调节因子谷胱甘肽过氧化物酶4(glutathione peroxidase 4,GPX4)蛋白表达。马松三色染色(Masson′s trichrome staining, MASSON)法分析各组大鼠肝组织病理改变。逆转录聚合酶链反应(reverse transcription polymerase chain reaction, RT-PCR)法检测各组大鼠肝组织中PIK3、Akt、mTOR的mRNA水平。结果 与对照组比较,DEN模型Objective To explore the regulatory effects of emodin on the phosphoinositide 3-kinase(PI3K)/protein kinase B(Akt)/mammalian target of ra-pamycin(mTOR)signaling pathway in diethylnitrosamine(DEN)induced precancerous lesions of liver cancer of rats and its impacts on ferroptosis.Methods Modeling of precancerous lesions of liver cancer was carried out by DEN inducing precancerous lesions of liver cancer on rats.A total of 50 male SD rats were randomly divided into control group[no mod-eling+10 ml/(kg·d)normal saline by gavage],DEN model group[precancerous lesions of liver cancer modeling+10 ml/(kg·d)normal saline by gavage],DEN+emodin group[precancerous lesions of liver cancer modeling+80 mg/(kg·d)emodin concentrated solution by gavage],DEN+liver-protecting tablet group[precancerous lesions of liver canc-er modeling+900 mg/kg liver-protecting tablet by gavage]and DEN+emodin+liver-protecting tablet group[precan-cerous lesions of liver cancer modeling+80mg/(kg·d)emodin concentrated solution by gavage+900 mg/kg liver-pro-tecting tablet by gavage],with 10 rats in each.The rats in each group were treated continuously for 12 weeks.Serum ala-nine aminotransferase(ALT),aspartate aminotransferase(AST)and albumin(ALB)levels of rats in each group were detected by biochemical analysis;the liver indexes of rats in each group was compared.The levels of serum interleukin 1 beta(IL-1β),tumor necrosis factor alpha(TNF-α)and IL-10 contents of rats in each group were detected by enzyme-linked immunosorbent assay(ELISA).The pathological changes of liver tissues were observed by hematoxylin-eosin(HE)staining;the apoptosis of liver tissue cells was analyzed by terminal deoxynucleotidyl transferase mediated dUTP nick-end labeling(TUNEL).The protein expression of glutathione peroxidase 4(GPX4),a central regulator of ferropto-sis,in liver tissues of rats in each group was detected by immunohistochemistry.The pathological changes in liver tissues of rats in each group were analyzed by Masson's trichrome staining(MASSON).The mRNA level

关 键 词：大黄素 肝癌前病变 磷脂酰肌醇3-激酶/蛋白激酶B/哺乳动物雷帕霉素靶蛋白信号通路 铁死亡 

分 类 号：R285[医药卫生—中药学] R735.7[医药卫生—中医学]