摘要
目的:通过腺嘌呤诱导的慢性肾脏病(CKD)大鼠模型结合体外实验,探讨铁死亡在肾脏纤维化中的作用和机制。方法:5~6周龄雄性SPF级SD大鼠(190~230 g)随机分为对照组(n=15)和CKD组(n=18),通过腺嘌呤灌胃法构建CKD模型。分别于第2、4、6周末,每组随机选取5只大鼠,处死并留取肾组织,检测铁、丙二醛(MDA)及谷胱甘肽(GSH)含量,逆转录-聚合酶链反应(RT-PCR)、免疫组化检测肾脏铁死亡及纤维化相关因子表达。体外实验用铁死亡诱导剂(RSL3)诱导人肾小管上皮细胞(HK-2细胞)死亡,铁死亡抑制剂(Fer-1)抑制细胞死亡,检测经干预后的细胞活力、铁死亡和纤维化相关因子表达。结果:CKD组大鼠肾组织铁、MDA升高,GSH降低,α平滑肌肌动蛋白(α-SMA),I型胶原蛋白(COL1A1)表达均升高,转铁蛋白受体1(TFR-1)和铁转运蛋白(FPN)表达先升高后降低,铁蛋白(FTH)表达均升高,4-羟基壬烯醛(4-HNE)表达升高,谷胱甘肽过氧化物酶4(GPX4)表达均降低,上述改变随时间进展呈现动态变化。GPX4表达与α-SMA、COL1A1呈负相关。RSL3处理的HK-2细胞表现为细胞活力降低、活性氧(ROS)升高、GSH含量减少、GPX4表达降低,α-SMA和COL1A1的表达升高,上述改变可被Fer-1逆转。结论:腺嘌呤诱导的CKD大鼠肾脏纤维化进程中均存在铁死亡,且促进纤维化的进展,可能与铁死亡刺激肾小管上皮细胞转分化有关。
Objective:To explore the role and mechanism of ferroptosis in renal fibrosis through an adenine-induced rat model of CKD combined with in vitro experiments.Methodology:5~6-week-old male SPF-grade SD male rats(190~230 g)rats were randomly divided into Control group(n=15)and CKD group(n=18),and the CKD model was constructed by adenine gavage.At the end of the 2nd,4th and 6th weekends,5 rats were randomly selected from each group to be executed,and iron content,malondialdehyde(MDA)content and glutathione(GSH)content were detected.RT-PCR and immunohistochemistry were used to detect the expression of renal ferroptosis and fibrosis-associated factors.In vitro experiments,ferroptosis was induced in human renal tubular epithelial cells(HK-2)by ferroptosis inducer(RSL3)and inhibited by ferroptosis inhibitor(Ferrostatin-1,Fer-1).Cell viability,ferroptosis-related markers,and fibrosis-associated factors were measured after intervention.Results:The CKD group showed increased iron content,MDA content,and decreased GSH levels.The expression levels ofα-smooth muscle actin(α-SMA)and alpha 1 type I collagen(COL1A1)increased,transferrin receptor 1(TFR-1)and ferroportin(FPN)initially increased and then decreased;ferritin heavy chain(FTH)and 4-hydroxynonenal(4-HNE)levels increased;and GPX4 expression decreased.The above changes showed dynamic progression over time.GPX4 expression level was negatively correlated withα-SMA and COL1A1 levels.RSL3-treated HK-2 cells exhibited reduced viability,increased reactive oxygen species(ROS),decreased GSH content and GPX4 expression,and upregulatedα-SMA and COL1A1.The above changes could be reversed by Fer-1.Conclusion:Ferroptosis was involved in the fibrotic process of adenine-induced CKD rats and contributes to the progression of fibrosis,which may be related to ferroptosis-stimulated transdifferentiation of renal tubular epithelial cells.
作者
刘义琴
朱婷婷
毛海霞
康婷
张丽玲
吴蔚桦
欧三桃
LIU Yiqin;ZHU Tingting;MAO Haixia;KANG Ting;ZHANG Liling;WU Weihua;OU Santao(Department of Nephrology,the Affiliated Hospital of Southwest Medical University,Sichuan Clinical Medical Research Centre for Nephrology,Sichuan Key Laboratory of Metabolic Vascular Diseases,Luzhou 646000,China)
出处
《肾脏病与透析肾移植杂志》
2025年第4期335-341,共7页
Chinese Journal of Nephrology,Dialysis & Transplantation
基金
四川省医学会慢病专项科研项目(2024HR140)。
关键词
慢性肾脏病
肾脏纤维化
铁死亡
chronic kidney disease
renal fibrosis
ferroptosis
