摘要
目的研究梓醇扶正制毒配伍对雷公藤甲素所致肝损伤的减毒作用及其铁死亡相关机制。方法通过动物实验筛选雷公藤甲素所致小鼠肝毒性的最佳剂量,再将20只C57BL/6J小鼠分为空白组、雷公藤甲素组(0.6 mg/kg)、梓醇组(4.5 mg/kg)和两药配伍组(0.6 mg/kg+4.5 mg/kg),5只/组。灌胃给药13 d后,检测小鼠肝组织病理、肝功能指标、亚细胞结构、脂质过氧化指标、亚铁离子沉积和铁死亡相关蛋白。在肝HL7702细胞中,采用铁死亡抑制剂Fer-1(1μmol/L)抑制铁死亡后,检测肝功能、Fe2+浓度和脂质过氧化;再将肝HL7702细胞分为空白组、梓醇组(80μg/L)、雷公藤甲素组(20μg/L)和两药配伍组(80μg/L+20μg/L),检测肝功能指标、Fe2+浓度、脂质过氧化、ROS水平和铁死亡相关蛋白,进一步验证铁死亡在梓醇减轻雷公藤甲素肝毒性中的作用。结果梓醇可减轻雷公藤甲素所致小鼠肝组织病理损伤并降低ALT、AST和LDH(P<0.01),同时逆转雷公藤甲素所致的Fe2+浓度和MDA的升高、GPX的降低(P<0.05)。Fer-1抑制铁死亡逆转了雷公藤甲素所致肝HL7702细胞ALT、AST和LDH的升高(P<0.01)。Western blotting和qRT-PCR结果显示,梓醇能逆转雷公藤甲素所致的铁死亡相关SLC7A11、FTH1、GPX4蛋白和基因表达(P<0.05)。结论梓醇通过SLC7A11/GPX4通路抑制过度铁死亡减轻雷公藤甲素的肝毒性。
Objective To investigate the protective effect of catalpol against triptolide-induced liver injury and explore its mechanism to test the Fuzheng Zhidu theory for detoxification.Methods C57BL/6J mice were randomized into blank control group,catalpol group,triptolide group and triptolide+catalpol group.After 13 days of treatment with the agents by gavage,the mice were examined for liver tissue pathology,liver function,hepatocyte subcellular structure,lipid peroxidation,ferrous ion deposition and expressions of ferroptosis-related proteins in the liver.In a liver cell line HL7702,the effect of catalpol or the ferroptosis inhibitor Fer-1 on triptolide-induced cytotoxicity was tested by examining cell functions,Fe2+concentration,lipid peroxidation,ROS level and the ferroptosis-related proteins.Results In C57BL/6J mice,catalpol significantly alleviated triptolide-induced hepatic injury,lowered the levels of ALT,AST and LDH,and reversed the elevation of Fe2+concentration and MDA level and the reduction of GPX level.In HL7702 cells,inhibition of ferroptosis by Fer-1 significantly reversed triptolide-induced elevation of ALT,AST and LDH levels.Western blotting and qRT-PCR demonstrated that catalpol reversed abnormalities in expressions of SLC7A11,FTH1 and GPX4 at both the mRNA and protein levels in triptolide-treated HL7702 cells.Conclusion The combined use of catalpol can reduce the hepatotoxicity of triptolide in mice by inhibiting excessive hepatocyte ferroptosis through the SLC7A11/GPX4 pathway.
作者
张林落
李长青
皇玲玲
周学平
娄媛媛
ZHANG Linluo;LI Changqing;HUANG Lingling;ZHOU Xueping;LOU Yuanyuan(Rehabilitation College in Jiangsu College of Nursing,Huai'an 223000,China;First Clinical School of Nanjing University of Chinese Medicine,Nanjing 210009,China;Department of Traditional Chinese Medicine,Affiliated Huai'an First People's Hospital of Nanjing Medical University,Huai'an 223300,China)
出处
《南方医科大学学报》
北大核心
2025年第4期810-818,共9页
Journal of Southern Medical University
基金
中国中医科学院学部委员学术传承与传播专项项目(CL2022E028XB)
江苏省高校自然科学研究课题面上项目(23KJD330001)
江苏省中医药科技发展计划课题面上项目(MS2024136)
淮安市基础研究计划自然科学研究课题面上项目(HAB2024076)
江苏高校“青蓝工程”
江苏护理职业学院科研创新研究团队项目(SHCXTD2023052904)。
关键词
梓醇
雷公藤甲素
肝毒性
铁死亡
catalpol
triptolide
hepatotoxicity
ferroptosis
作者简介
通信作者:张林落,博士,讲师,E-mail:13674991073@163.com。
