首页|铁死亡在骨髓间充质干细胞调控肝纤维化中的作用

铁死亡在骨髓间充质干细胞调控肝纤维化中的作用

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目的 探讨铁死亡在骨髓间充质干细胞(BMMSCs)调控肝星状细胞(HSCs)活化和肝纤维化中的作用.方法 选取5只SD雄性大鼠(北京军事医学科学院动物实验中心提供)腹腔注射橄榄油12周作为对照组;选取15只SD大鼠腹腔注射含40%CCl4的橄榄油8周建立大鼠肝纤维化模型,采用随机数表法分为模型组、BMMSCs治疗组和小檗碱组,每组5只;将大鼠肝星状细胞(HSC-T6)分为对照组(Ctrl)、血小板衍生因子(PDGF-BB)激活组、PDGF-BB+BMMSCs共培养组、PDGF-BB+爱拉斯汀(erastin)组及PDGF-BB+BMMSCs+铁死亡抑制剂(fer-1)组.通过苏木精-伊红和天狼猩红染色评估肝脏病理,检测不同组别的丙二醛(MDA)和还原型谷胱甘肽(GSH)含量,通过蛋白质印迹法(Western blot)和免疫荧光检测细胞纤维化和铁死亡指标水平.两组间数据比较采用独立样本t检验;3组及以上数据比较采用单因素方差分析.结果 BMMSCs组肝细胞变性、坏死及胶原纤维沉积明显减轻;BMMSCs共培养组α-平滑肌肌动蛋白(α-SMA)表达水平低于PDGF-BB激活组(0.80±0.10 比 1.13±0.06,F=32.914,P<0.05).BMMSCs 组大鼠肝脏 MDA 含量高于模型组[(0.37±0.02)μmol/mg 比(0.23±0.02)μmol/mg,F=56.360,P<0.05].BMMSCs 共培养组 MDA含量高于 PDGF-BB 激活组[(1.54±0.18)nmol/mg prot 比(1.14±0.06)nmol/mg prot,F=16.397,P<0.05],GSH 含量低于 PDGF-BB 激活组[(0.63±0.05)μg/106 cells 比(0.96±0.03)μg/106 cells,F=50.638,P<0.05].fer-1预处理组细胞活力高于BMMSCs共培养组[(78.50±2.73)%比(24.07±8.92)%,F=89.533,P<0.05],并且fer-1抑制BMMSCs介导的诱导细胞发生铁死亡和减轻纤维化作用.结论 铁死亡参与BMMSCs抑制HSCs活化和减轻肝纤维化过程.
Role of ferroptosis in regulating liver fibrosis inhibition by bone marrow mesenchymal stem cells
Objective To investigate the role of ferroptosis in the regulation of hepatic stellate cells activation and liver fibrosis by bone marrow mesenchymal stem cells.Methods In total,5 male SD rats(provided by the Animal Experimental Center of Beijing Academy of Military Medical Sciences)were intra-peritoneally injected with olive oil for 12 weeks as the control group,and 15 SD rats were intraperitoneally injected with olive oil containing 40%CC14 for 8 weeks to establish rat liver fibrosis model,and were divid-ed into model group,BMMSCs treatment group and berberine group,5 per group.Rat hepatic stellate cells were divided into control group(Ctrl),platelet-derived growth factor BB(PDGF-BB)activation group,PDGF-BB+BMMSCs co-culture group,PDGF-BB+erastin group and PDGF-BB+BMMSCs+fer-1 group.Liver pathology was evaluated by hematoxylin and eosin(HE)and Sirius red staining,and the levels of ma-londialdehyde(MDA)and glutathione(GSH)were detected.Fibrosis and ferroptosis indicators were detec-ted by Western blotting and immunofluorescence.Data between two groups were compared by independent sample t test;data between three groups and above were analyzed by one-way analysis of variance.Results The degeneration,necrosis and collagen fiber deposition of hepatocytes in BMMSCs group were significantly reduced;the expression level of α-smooth muscle actin(α-SMA)in BMMSCs co-culture group was lower than PDGF-BB activation group(0.80±0.10 vs.1.13±0.06,F=32.914,P<0.05).The MDA con-tent in BMMSCs group was higher than model group[(0.37±0.02)vs.(0.23±0.02)μmoVmg,F=56.360,P<0.05].The MDA content of BMMSCs co-culture group was higher than PDGF-BB activation group[(1.54±0.18)vs.(1.14±0.06)nmol/mg prot,F=16.397,P<0.05],and the GSH content was lower than PDGF-BB activation group[(0.63±0.05)vs.(0.96±0.03)μg/106 cells,F=50.638,P<0.05].The cell viability of fer-1 pretreatment group was higher than BMMSCs co-culture group[(78.50±2.73)%vs.(24.07±8.92)%,F=89.533,P<0.05],and fer-1 inhibited BMMSCs-media-ted induction of ferroptosis and alleviated fibrosis.Conclusion Ferroptosis is involved in the process of BMMSCs inhibiting HSCs activation and alleviating liver fibrosis.

Bone marrow mesenchymal stem cellsLiver fibrosisFerroptosis

张新如、王玉鑫、左怀文、皮艺林、周慧媛、邓腊梅、宋红丽

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天津医科大学一中心临床学院,天津 300192

南开大学医学院,天津 300074

天津市第一中心医院肝移植科天津市器官移植重点实验室天津市器官移植临床医院研究中心,天津 300192

骨髓间充质干细胞 肝纤维化 铁死亡

国家自然科学基金

82070639

2024

中华实验外科杂志
中华医学会

中华实验外科杂志

CSTPCD
影响因子:0.759
ISSN:1001-9030
年,卷(期):2024.41(6)
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