摘要
目的探讨血红素加氧酶1(HO-1)修饰的骨髓间充质干细胞(BMMSCs)对缺氧/复氧(H/R)诱导的脂肪变肝细胞损伤的作用及其机制。方法将脂肪变大鼠肝细胞(IAR20)按照实验要求分为对照组(Ctrl)、H/R组、H/R+铁死亡抑制剂组(Fer-1)、铁死亡诱导剂组(Erastin)、H/R+BMMSCs(B)组、H/R+HO-1/BMMSCs(HB)组、H/R+HB+小干扰RNA阴性对照组(si-NC)、H/R+HB+敲降GPX4组(si-GPX4)。通过检测不同组别细胞的脂质活性氧(Lipid ROS)、谷胱甘肽过氧化物酶4(GPX4)、丙二醛(MDA)、谷胱甘肽(GSH)的表达评价铁死亡的严重程度。两组和多组间的数据比较分别采用独立样本t检验和单因素方差分析。结果Fer-1及H/R+B处理组Lipid ROS与MDA的表达量均低于H/R组[(1.65±0.02)、(1.85±0.04)比(3.50±0.03),t=87.600、60.030,P<0.05;(154.50±0.84)%、(228.90±21.49)%比(676.50±22.36)%,t=36.380、25.000,P<0.05];Fer-1及H/R+B处理组GSH水平均高于H/R组[(0.75±0.03)、(0.72±0.04)比(0.45±0.01),t=19.400、17.060,P<0.05];H/R+HB组的Lipid ROS及MDA水平低于H/R+B组[(1.31±0.03)比(1.85±0.04),t=19.400,P<0.05;(150.10±32.82)%比(228.90±21.49)%,t=3.478,P<0.05],其GSH水平高于H/R+B组[(0.81±0.03)比(0.72±0.04),t=4.981,P<0.05]。si-GPX4组的Lipid ROS与MDA表达量均高于si-NC组[(2.87±0.13)比(1.36±0.06),t=18.100,P<0.05;(323.30±12.58)%比(184.30±7.09)%,t=16.670,P<0.05];si-GPX4组的GPX4与GSH表达量均低于si-NC组[(0.45±0.03)比(0.92±0.04),t=7.692,P<0.05;(0.54±0.07)比(0.75±0.15),t=5.076,P<0.05]。结论HO-1/BMMSCs通过促进GPX4的表达抑制铁死亡以修复受损的脂肪变肝细胞。
Objective To investigate the effect and mechanism of heme oxygenase 1(HO-1)-modified bone marrow mesenchymal stem cells(BMMSCs)on hypoxia/reoxygenation(H/R)-induced steatotic hepatocytes injury.Methods Steatotic hepatocytes of rat(IAR20)were divided into control group(Ctrl),H/R group,H/R+ferroptosis inhibitor group(Fer-1),ferroptosis inducer group(Erastin),H/R+BMMSCs(B)group,H/R+HO-1/BMMSCs(HB)group,H/R+HB+siRNA negative control group(si-NC),H/R+HB+knockdown GPX4 group(si-GPX4).The expression levels of lipid ROS,GPX4,MDA and GSH in different groups of cells were measured to evaluate the severity of ferroptosis.Statistical data were compared between two groups and multiple groups using independent sample t-test and one-way ANOVA,respectively.Results The Lipid ROS and MDA expressions of Fer-1 and H/R+B groups were significantly lower than those of H/R group[(1.65±0.02),(1.85±0.04),vs.(3.50±0.03),t=87.600,60.030,P<0.05;(154.50±0.84)%,(228.90±21.49)%vs.(676.50±22.36)%,t=36.380,25.000,P<0.05].The levels of GSH in Fer-1 and H/R+B treatment groups were higher than those in H/R group[(0.75±0.03),(0.72±0.04)vs.(0.45±0.01),t=19.400,17.060,P<0.05].Lipid ROS and MDA levels in H/R+HB group were significantly lower than those in H/R+B group[(1.31±0.03)vs.(1.85±0.04),t=19.400,P<0.05;(150.10±32.82)%vs.(228.90±21.49)%,t=3.478,P<0.05],GSH level was higher than that of H/R+B group[(0.81±0.03)vs.(0.72±0.04),t=4.981,P<0.05].The expression levels of lipid ROS and MDA in si-GPX4 group were higher than those in si-NC group[(2.87±0.13)vs.(1.36±0.06),t=18.100,P<0.05;(323.30±12.58)%vs.(184.30±7.09)%,t=16.670,P<0.05].The expression levels of GPX4 and GSH in si-GPX4 group were lower than those in si-NC group[(0.45±0.03)vs.(0.92±0.04),t=7.692,P<0.05;(0.54±0.07)vs.(0.75±0.15),t=5.076,P<0.05].Conclusion HO-1/BMMSCs can inhibit ferroptosis to repair damaged steatotic hepatocytes by promoting the expression of GPX4.
作者
田小荣
吴龙龙
左怀文
袁梦淑
王玉鑫
张新如
宋红丽
Tian Xiaorong;Wu Longlong;Zuo Huaiwen;Yuan Mengshu;Wang Yuxin;Zhang Xinru;Song Hongli(The First central Clinical College of Tianjin Medical University,Tianjin 300192,China;The Medical College,Naikai University,Tianjin 300074,China;Department of Organ Transplantation,Tianjin First Central Hospital,Tianjin Key Laboratory of Organ Transplantation,Tianjin Clinical Medicine Research Center for Organ Transplantation,Tianjin 300192,China)
出处
《中华实验外科杂志》
CAS
北大核心
2022年第10期1884-1887,共4页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金(82070639、81670574)。
关键词
骨髓间充质干细胞
脂肪变肝细胞
缺氧/复氧
铁死亡
Bone marrow mesenchymal stem cells
Steatotic hepatocytes
Hypoxia/reoxygenation
Ferroptosis
