中药新药与临床药理2026，Vol.37Issue(3)：404-412,9.DOI:10.19378/j.issn.1003-9783.2026.03.003
大黄素调控Tribbles同源蛋白3诱导小鼠肾小管上皮细胞氧化应激损伤的作用及机制
Effect and Mechanism of Emodin Inducing Oxidative Stress Injury in Murine Renal Tubular Epithelial Cells via TRIB3 Upregulation
摘要
Abstract
Objective To investigate the toxic effects of emodin on mouse kidneys and the underlying molecular mechanisms based on Tribbles homolog 3(TRIB3).Methods ICR mice were randomly divided into a control group,a low-dose emodin group,and a high-dose emodin group,with 6 mice in each group.The low-dose and high-dose emodin groups received intraperitoneal injections of emodin at doses of 20 mg·kg-1 and 40 mg·kg-1,respectively,for 4 consecutive weeks.Mouse renal tubular epithelial cells(TCMK-1)were treated with emodin at final concentrations of 10,20,40,80,and 100 μmol·L-1 for 24 and 48 hours.Pathological and morphological changes in kidney tissue were observed by hematoxylin-eosin(HE),Masson,periodic acid-Schiff(PAS),and Sirius Red staining.Levels of malondialdehyde(MDA)and reduced glutathione(GSH)in kidney tissue were measured.Cell viability was detected by the CCK-8 assay.Intracellular reactive oxygen species(ROS)levels were detected using the fluorescent probes 2',7'-dichlorodihydrofluorescein diacetate(DCFH-DA)and dihydroethidium(DHE).The mRNA expression levels of TRIB3 and α-smooth muscle actin(α-SMA)in kidney tissue and TCMK-1 cells were measured by RT-qPCR.The protein expression level of TRIB3 in TCMK-1 cells was detected by Western Blot.The expression levels of 4-hydroxynonenal(4-HNE),α-SMA,kidney injury molecule-1(KIM-1),and TRIB3 in kidney tissue were determined by immunohistochemistry.Results(1)Compared with the control group,the kidneys of mice in the low-dose and high-dose emodin groups showed significant atrophy,with markedly decreased kidney indices(P＜0.05,P＜0.01),and significantly increased expression of KIM-1 in kidney tissue(P＜0.05,P＜0.01).The high-dose emodin group exhibited significant pathological changes,including edema of renal tubular epithelial cells,protein cast formation and vacuolar degeneration in tubular lumens,thickened basement membranes,glomerular enlargement with capsular changes,inflammatory cell infiltration in the renal interstitium,and interstitial fibrosis.(2)Compared with the control group,mice in the low-dose and high-dose emodin groups showed increased collagen fiber deposition around glomeruli and tubules,a significantly elevated collagen volume fraction(P＜0.01),more pronounced thickening of the glomerular basement membrane,mesangial hyperplasia,vacuolar changes in renal tubules,and inflammatory infiltration in the interstitium.The area of red-stained type I collagen fibers was significantly increased in kidney tissue.The mRNA and protein expression levels of α-SMA in the high-dose emodin group were significantly elevated(P＜0.05,P＜0.001).(3)Compared with the control group,the MDA content in kidney tissue was significantly increased(P＜0.01)and the GSH level was significantly decreased(P＜0.01)in the high-dose emodin group.The expression levels of 4-HNE in kidney tissue were significantly increased in both the low-dose and high-dose emodin groups(P＜0.05,P＜0.001).(4)After 24 and 48 hours of intervention,the viability of TCMK-1 cells in the 10-100 μmol·L-1 emodin groups was significantly reduced compared with the control group(0 μmol·L-1)(P＜0.01,P＜0.001).After 3 hours of intervention,the green or red fluorescent signals in TCMK-1 cells from the 20 and 40 μmol·L-1 emodin groups were significantly enhanced,indicating a significant increase in intracellular ROS accumulation.(5)Compared with the control group,the protein expression of TRIB3 in kidney tissue was significantly upregulated in both the low-dose and high-dose emodin groups(P＜0.05,P＜0.01),and the mRNA expression of TRIB3 was significantly upregulated in the high-dose emodin group(P＜0.05).The protein and mRNA expression of TRIB3 in TCMK-1 cells from the 20 and 40 μmol·L-1 emodin groups were significantly upregulated(P＜0.01,P＜0.001).Conclusion Long-term,high-dose administration of emodin can cause structural and functional damage to mouse kidneys.The underlying toxic mechanism may be related to inducing abnormal expression of TRIB3,leading to oxidative stress injury in renal tubular epithelial cells,which subsequently contributes to renal fibrosis.关键词
大黄素/肾毒性/Tribbles同源蛋白 3/氧化应激/肾纤维化/肾小管上皮细胞/小鼠Key words
emodin/nephrotoxicity/Tribbles homolog 3/oxidative stress/renal fibrosis/renal tubular epithelial cells/mice分类
医药卫生引用本文复制引用
冯中元,黄腾,鹿红,马源彪,韩炜钰,李佳琪,邓哲文,陆争新,张一沐,李佳齐,马博..大黄素调控Tribbles同源蛋白3诱导小鼠肾小管上皮细胞氧化应激损伤的作用及机制[J].中药新药与临床药理,2026,37(3):404-412,9.基金项目
国家自然科学基金项目(82405508) （82405508）
江苏省药品监督管理局药品监管科学科研计划项目(202406) （202406）
江苏省药品监督管理局科研计划项目(202309). （202309）
