摘要
目的通过蛋白质组学方法探讨消癌解毒方促进肝癌小鼠肿瘤细胞铁死亡,抑制移植瘤生长的作用机制。方法C57BL/6J小鼠随机分为模型组、消癌解毒方低剂量组、消癌解毒方高剂量组、消癌解毒方联合铁死亡抑制剂组、铁死亡抑制剂组、顺铂组。构建H22小鼠移植瘤模型,消癌解毒方低、高剂量组予消癌解毒方灌胃,剂量分别为(10、20 g·kg^(-1)·d^(-1));铁死亡抑制剂组予Liproxstatin-1腹腔注射,剂量为10 mg·kg^(-1)·d^(-1);顺铂组予顺铂腹腔注射,剂量为10 mg·kg^(-1)·d^(-1);消癌解毒方联合铁死亡抑制剂组予消癌解毒方(20 g·kg^(-1)·d^(-1))灌胃以及Liproxstatin-1(10 mg·kg^(-1)·d^(-1))腹腔注射,模型组灌胃等量生理盐水。连续给药11 d后剥取瘤体计算抑瘤率;HE染色检测病理变化;透射电镜观察线粒体结构变化;流式细胞术检测瘤体组织活性氧(ROS)水平;制备血清以TMT肽段标记结合LC-MS/MS寻找差异蛋白表达谱、应用IPA软件进行分析;生化检测血清铁离子、还原型谷胱甘肽(GSH)及丙二醛(MDA)含量;Western blot法检测核因子E2相关因子2(Nrf2)、血红素加氧酶-1(HMOX1)、胱氨酸/谷氨酸逆向转运蛋白(SLC7A11)和谷胱甘肽过氧化物酶4(GPX4)的蛋白表达水平。结果消癌解毒方低、高剂量及顺铂能抑制瘤体生长,抑瘤率分别为36.12%、51.63%、57.43%,铁死亡抑制剂促进了瘤体的生长,抑瘤率为-45.56%,消癌解毒方联合铁死亡抑制剂组瘤体较铁死亡抑制剂组缩小,抑瘤率为18.11%;HE染色显示消癌解毒方高剂量组、顺铂组瘤体组织出现凋亡细胞及大量空泡蓄积;透射电镜结果显示,消癌解毒方低、高剂量组出现一定程度的线粒体萎缩及膜密度增加;流式结果显示:消癌解毒方干预后ROS水平明显升高(P<0.01);蛋白质组学检测显示,消癌解毒方高剂量组与模型组相比差异蛋白共129个,其中下调蛋白62个,上调蛋白67个,差异表达涉及脂质代�
OBJECTIVE To investigate the mechanism of Xiaoai Jiedu Recipe in promoting ferroptosis of tumor cells and inhibiting the growth of transplanted tumors in hepatocellular carcinoma mice by proteomics method.METHODS C57BL/6J mice were randomly divided into,model group,low-and high-dose groups of Xiaoai Jiedu Recipe,Xiaoai Jiedu Recipe combined with ferroptosis inhibitor group,ferroptosis inhibitor group and cisplatin group.The H22 mouse transplantation tumor model was constructed and the drug administration interventions were as follows:the low-and high-dose groups of Xiaoai Jiedu Recipe were given Xiaoai Jiedu Recipe by gavage at the doses of 10 and 20 g·kg^(-1)·d^(-1);the ferroptosis inhibitor group was given Liproxstatin-1 by intraperitoneal injection at the dose of 10 mg·kg^(-1)·d^(-1);the cisplatin group was given cisplatin by intraperitoneal injection at the dose of 10 mg·kg^(-1)·d^(-1);the Xiaoai Jiedu Recipe combined with ferroptosis inhibitor group was given a gavage dose of 20 g·kg^(-1)·d^(-1)of Xiaoai Jiedu Recipe and an intraperitoneal injection of 10 mg·kg^(-1)·d^(-1)of the ferroptosis inhibitor Liproxstatin-1;the model group was given an equal amount of saline by gavage.The drugs were administered for 11 d continuously.The tumors were stripped to calculate tumor inhibition rate.Pathological changes were observed by hematoxylin-eosin(HE).Mitochondrial structural changes were observed by transmission electron microscopy.Reactive oxygen species(ROS)levels were detected by flow cytometry.Serum was prepared and analysed by TMT peptide labelling combined with LC-MS/MS to find differential protein expression profiles,applying IPA software.Serum iron ions,glutathione(GSH)and malondialdehyde(MDA)levels were measured biochemically.Protein expression levels of nuclear factor E2-related factor 2(Nrf2),heme oxygenase-1(HMOX1),cystine/glutamate reverse transporter protein(SLC7A11)and glutathione peroxidase 4(GPX4)were measured by protein Western blot.RESULTS The tumor growth was inhibited in the low-and hi
作者
李文婷
张淇
吴勉华
李黎
姜泽群
张羽
杨卫豪
费凡
LI Wen-ting;ZHANG Qi;WU Mian-hua;LI Li;JIANG Ze-qun;ZHANG Yu;YANG Wei-hao;FEI Fan(The First School of Clinical Medicine,Nanjing University of Chinese Medicine,Nanjing 210023,China;Jiangsu Collaborative Innovation Center of Traditional Chinese Medicine Prevention and Treatment of Tumor,Nanjing 210023,China;School of Medicine&Holistic Integrative Medicine,Nanjing University of Chinese Medicine,Nanjing 210023,China)
出处
《南京中医药大学学报》
CAS
CSCD
北大核心
2023年第12期1179-1188,共10页
Journal of Nanjing University of Traditional Chinese Medicine
基金
国家自然科学基金青年科学基金项目(81804058)
第二届全国名中医传承工作室(国中医药办人教函〔2022〕245号)
吴勉华全国名老中医药专家传承工作室(国中医药人教函〔2022〕75号)
国家中医药管理局第七批全国老中医药专家学术经验继承工作项目(国中医药人教函〔2022〕76号)
吴勉华江苏省名老中医药专家传承工作室(苏中医科教〔2021〕7号)
国家中医药管理局中医药传承与创新“百千万”人才工程(岐黄工程)岐黄学者项目(国中医药人教函〔2018〕284号)
国家级大学生创新训练计划项目(202110315012)。
