中国中药杂志2026，Vol.51Issue(8)：2310-2322,13.DOI:10.19540/j.cnki.cjcmm.20251208.801
橙皮素通过靶向线粒体调控肝细胞焦亡在缓解酒精性肝损伤中的作用及机制
Effects and mechanisms of hesperetin in attenuating alcoholic liver injury by targeting mitochondria to regulate hepatocyte pyroptosis
摘要
Abstract
This study investigated the effects and potential mechanisms of hesperetin(HST)in targeting mitochondria to regulate hepatocyte pyroptosis and alleviate alcoholic liver injury through in vitro experiments.Human immortalized hepatocytes(THLE-2)were used to establish an in vitro alcoholic liver injury model induced by 200 mmol·L-1 ethanol.The effective concentrations of HST(40,80,and 160 μmol·L-1)were determined using the MTT assay.The effects of HST on hepatocyte pyroptosis were evaluated by optical microscopy,lactate dehydrogenase(LDH)release assay,and flow cytometry,and its effects on lipid accumulation were assessed using Nile red staining and cholesterol and triglyceride assay kits.Further intervention with a high dose of HST was performed,and the expression levels of pyroptosis-related molecules,including caspase-1,gasdermin D(GSDMD),interleukin-18(IL-18),and interleukin-1β(IL-1β),were detected by Western blot and PCR.In addition,a caspase-1 inhibitor(Z-YVAD-FMK)was used to reversely validate the regulatory effect of HST on the classical pyroptosis pathway.The morphology of the endoplasmic reticulum,lysosomes,and mitochondria was observed using confocal laser scanning microscopy.Superoxide dismutase(SOD)levels were measured using a commercial assay kit,mitochondrial membrane potential was analyzed by flow cytometry,and the gene expression levels of mitochondrial membrane transport proteins,including voltage-dependent anion channel 1(VDAC1),voltage-dependent anion channel 2(VDAC2),translocase of the outer mitochondrial membrane 20(TOM20),and translocase of the outer mitochondrial membrane 34(TOM34),were determined by PCR.Meanwhile,the mitochondrial reactive oxygen species(mtROS)inducer rotenone was employed to validate the mitochondria-targeted regulatory effects of HST on mitochondrial function.The experimental results showed that HST concentration-dependently increased hepatocyte viability,reduced LDH levels,significantly alleviated hepatocyte pyroptosis,and improved lipid accumulation,while downregulating the expression of caspase-1,GSDMD,IL-18,and IL-1β.After the addition of a caspase-1 inhibitor,the inhibitory effect of HST on hepatocyte pyroptosis was further confirmed.Mechanistic investigations revealed that HST targeted mitochondria by scavenging mtROS,upregulating SOD expression,restoring mitochondrial membrane potential,and downregulating the expression of mitochondrial membrane-associated transport proteins VDAC1,VDAC2,TOM20,and TOM34.Furthermore,the addition of the mtROS inducer rotenone reversely validated the role of HST in alleviating hepatocyte pyroptosis through mitochondrial targeting.In summary,HST alleviates hepatocyte pyroptosis and improves lipid metabolism disorders by targeting mitochondria and regulating the classical pyroptosis pathway,thereby mitigating alcoholic liver injury.These findings provide a theoretical basis for the potential clinical application of HST in the treatment of alcoholic liver injury.关键词
橙皮素/酒精性肝损伤/细胞焦亡/线粒体靶向/脂代谢Key words
hesperetin/alcoholic liver injury/pyroptosis/mitochondria-targeted/lipid metabolism引用本文复制引用
张卫东,华丰,赵权,黄玮,张晶,王访,王飞虾,陈薇,张晓鸣..橙皮素通过靶向线粒体调控肝细胞焦亡在缓解酒精性肝损伤中的作用及机制[J].中国中药杂志,2026,51(8):2310-2322,13.基金项目
国家中药炮制技术传承基地建设项目(财社[2022]45号) （财社[2022]45号）
全国老药工传承工作室建设项目(国中医药人教函[2024]255号) （国中医药人教函[2024]255号）
江苏省老药工传承工作室建设项目(苏中医科教[2024]4号) （苏中医科教[2024]4号）
江苏省中药骨干人才高级研修项目(苏中医科教[2022]11号) （苏中医科教[2022]11号）
常州市第五批科技计划(社会发展科技支撑)项目(CE20235068) （社会发展科技支撑）
常州市科技计划(应用基础研究指导性)项目(CJ20229029,CJ20239042,CJ20242049,CJ20253125,CJ20219020,CJ20219011) （应用基础研究指导性）
常州市"龙城强医"工程中医药人才培育项目-张冰岐黄学者工作室项目(常财社[2024]55号) （常财社[2024]55号）
2024年度省中医药学会科研项目(苏中会[2024]62号) （苏中会[2024]62号）
2024年度常州市卫健委科技项目(WZ202405) （WZ202405）
江苏省中医疫病研究中心开放课题(JSYB2024KF30) （JSYB2024KF30）
南京中医药大学自然科学基金项目(XZR2024162) （XZR2024162）
江苏省自然科学基金青年项目(BK20230758) （BK20230758）
