摘要
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/10^(6) cells比(0.96±0.03)μg/10^(6) 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。
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 intraperitoneally injected with olive oil for 12 weeks as the control group,and 15 SD rats were intraperitoneally injected with olive oil containing 40%CCl4 for 8 weeks to establish rat liver fibrosis model,and were divided 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 malondialdehyde(MDA)and glutathione(GSH)were detected.Fibrosis and ferroptosis indicators were detected 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 ofα-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 content in BMMSCs group was higher than model group[(0.37±0.02)vs.(0.23±0.02)μmol/mg,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/10^(6) 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-mediated induction of ferroptosis and alleviated fibrosis.Conclusion Ferroptosis is involved in the process of BMMSCs inhibiting HSCs activation and alleviating liver fibrosis.
作者
张新如
王玉鑫
左怀文
皮艺林
周慧媛
邓腊梅
宋红丽
Zhang Xinru;Wang Yuxin;Zuo Huaiwen;Pi Yilin;Zhou Huiyuan;Deng Lamei;Song Hongli(The First Central Clinical College of Tianjin Medical University,Tianjin 300192,China;The Medical College,Nankai University,Tianjin 300074,China;Department of Liver Transplantation,Tianjin First Central Hospital,Tianjin Clinical Medical Research Center of Organ Transplantation,Tianjin 300192,China)
出处
《中华实验外科杂志》
CAS
2024年第6期1200-1204,共5页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金(82070639)。
关键词
骨髓间充质干细胞
肝纤维化
铁死亡
Bone marrow mesenchymal stem cells
Liver fibrosis
Ferroptosis
作者简介
通信作者:宋红丽,Email:hlsong26@163.com。
