切换至 "中华医学电子期刊资源库"
高级检索
中华肝脏外科手术学电子杂志

中华肝脏外科手术学电子杂志 ›› 2020, Vol. 09 ›› Issue (04) : 374 -379. doi: 10.3877/cma.j.issn.2095-3232.2020.04.018

所属专题： 文献；

基础研究

过表达Nrf2人脐带间充质干细胞对小鼠肝脏缺血-再灌注损伤的保护作用
刘荣强1, 林国桢2, 代天星2, 邓铭彬2, 周朝荣3, 汪国营2,†()   
  1. 1. 510630　广州，中山大学附属第三医院肝脏外科肝移植中心；510030　广州医科大学附属第一医院肝胆外科
    2. 510630　广州，中山大学附属第三医院肝脏外科肝移植中心
    3. 510220　广州医科大学附属第二医院普通外科
  • 收稿日期:2020-04-18 出版日期:2020-08-10
  • 通信作者: 汪国营
  • 基金资助:
    国家自然科学基金(81470870，81670601，81770648); 十三五科技重大专项(2017ZX10203205-006-001); 广东省自然科学基金(2016A030313278，2015A030313038，2015A030312013); 广东省科技计划项目(2014A020211015，2017B020209004); 广州市科技计划项目(2014J4100183)

Protection of Nrf2-overexpressing human umbilical cord mesenchymal stem cells on hepatic ischemia-reperfusion injury in mice

Rongqiang Liu1, Guozhen Lin2, Tianxing Dai2, Mingbin Deng2, Chaorong Zhou3, Guoying Wang2,†()   

  1. 1. Department of Liver Surgery, Liver Transplant Center, the Third Affiliated Hospital of Sun Yat-sen University, Guangzhou 510630, China; Department of Hepatobiliary Surgery, the First Affiliated Hospital of Guangzhou Medical University, Guangzhou 510030, China
    2. Department of Liver Surgery, Liver Transplant Center, the Third Affiliated Hospital of Sun Yat-sen University, Guangzhou 510630, China
    3. Department of General Surgery, the Second Affiliated Hospital of Guangzhou Medical University, Guangzhou 510220, China
  • Received:2020-04-18 Published:2020-08-10
  • Corresponding author: Guoying Wang
  • About author:
    Corresponding author: Wang Guoying, Email:
全文 ( ) 下载PDF ( ) 导出引用
引用本文：

刘荣强, 林国桢, 代天星, 邓铭彬, 周朝荣, 汪国营. 过表达Nrf2人脐带间充质干细胞对小鼠肝脏缺血-再灌注损伤的保护作用[J]. 中华肝脏外科手术学电子杂志, 2020, 09(04): 374-379.

Rongqiang Liu, Guozhen Lin, Tianxing Dai, Mingbin Deng, Chaorong Zhou, Guoying Wang. Protection of Nrf2-overexpressing human umbilical cord mesenchymal stem cells on hepatic ischemia-reperfusion injury in mice[J]. Chinese Journal of Hepatic Surgery(Electronic Edition), 2020, 09(04): 374-379.

目的

探讨过表达Nrf2的人脐带间充质干细胞（HUCMSCs）对小鼠缺血-再灌注损伤（IRI）的保护作用。

方法

制作70%肝脏IRI模型，HUCMSCs细胞分离培养、鉴定，构建过表达Nrf2慢病毒。正常SPF级健康雄性C57小鼠40只，按随机数字表法随机分为4组，分别为Sham组、IRI组、IRI+GFP-HUCMSCs、IRI+Nrf2-HUCMSCs组，缺血90 min后IRI+GFP-HUCMSCs、IRI+Nrf2-HUCMSCs组分别应用1×106 GFP-HUCMSCs、Nrf2-HUCMSCs 100 μl细胞悬液从小鼠下腔静脉注射，再灌注12、24 h后处死小鼠，下腔静脉取血检测ALT、AST；取肝脏组织行HE和TUNEL染色观察肝损伤和细胞凋亡情况。各组小鼠转氨酶比较采用单因素分析和LSD-t检验。

结果

再灌注12 h，IRI+Nrf2-HUCMSCs组肝细胞损伤较IRI、GFP-HUCMSCs组明显减轻；再灌注24 h，IRI+Nrf2-HUCMSCs组炎症细胞浸润退去，恢复也较快。再灌注12 h，IRI+Nrf2-HUCMSCs组肝功能ALT、AST分别为（1 472±540）、（1 592±596）U/L，明显低于IRI+GFP-HUCMSCs组的（4 700±1 526）、（5 464±1 210）U/L和IRI组的（7 452±1 637）、（6 928±2 439）U/L（LSD-t=-4.460，-7.757和-6.420，-4.753；P<0.05）。再灌注12 h，IRI+Nrf2-HUCMSCs组肝细胞凋亡率为（40±6）%，明显低于IRI+GFP-HUCMSCs组的（79±6）%和IRI组的（103±6）%（LSD-t=-8.162，-13.040；P<0.05）。

结论

过表达Nrf2的HUCMSCs对正常小鼠肝脏IRI可能是通过减少肝细胞凋亡发挥一定的保护作用。

关键词: 间充质干细胞, 核因子E2相关因子2（Nrf2）, 缺血-再灌注损伤, 肝损伤, 细胞凋亡, 小鼠
Objective

To explore the protection of Nrf2-overexpressing human umbilical cord mesenchymal stem cells (HUCMSCs) on hepatic ischemia-reperfusion injury (IRI) in mice.

Methods

A model of partial (70%) hepatic IRI was constructed. HUCMSCs were isolated, cultured and identified, and Nrf2-overexpressing lentiviral was constructed. 40 healthy male C57 mice of SPF grade were randomly divided into 4 groups according to the random number table: Sham group, IRI group, IRI+GFP-HUCMSCs group, and IRI+Nrf2-HUCMSCs group. For IRI+GFP-HUCMSCs and IRI+Nrf2-HUCMSCs group, 1×106 GFP-HUCMSCs, and 100 μl of Nrf2-HUCMSCs cell suspension were injected in the inferior vena cava of mice respectively after 90 min of ischemia. The mice were executed after 12 or 24 h of reperfusion, and then blood was taken from the inferior vena cava for ALT and AST test. Liver injury and cell apoptosis were observed through HE and TUNEL staining of liver tissue. Transaminases levels of each group were compared using univariate analysis and LSD-t test.

Results

After 12 h of reperfusion, hepatocyte damage was significantly milder in the IRI+Nrf2-HUCMSCs group than that in the IRI and GFP-HUCMSCs groups. After 24 h of reperfusion, inflammatory cell infiltration receded and recovered more quickly in the IRI+Nrf2-HUCMSCs group. After 12 hof reperfusion, the ALT and AST levels were respectively (1 472±540), (1 592±596) U/L in the IRI+Nrf2-HUCMSCs group, which were significantly lower than (4 700±1 526), (5 464±1 210) U/L in the IRI+GFP-HUCMSCs group, and (7 452±1 637), (6 928±2 439) U/L in the IRI group (LSD-t=-4.460, -7.757 and -6.420, -4.753; P<0.05). After 12 h of reperfusion, the rate of hepatocyte apoptosis was (40±6)% in the IRI+Nrf2-HUCMSCs group, significantly lower than (79±6)% in the IRI+GFP-HUCMSCs group and (103±6)% in the IRI group (LSD-t=-8.162, -13.040; P<0.05).

Conclusions

HUCMSCs overexpressing Nrf2 may exert certain protective effects on hepatic IRI by reducing the hepatocyte apoptosis in healthy mice.

Key words: Mesenchymal stem cells, Nuclear factor E2-related factor 2 (Nrf2), Ischemia-reperfusion injury, Liver injury, Apotosis, Mouse
图1 HUCMSCs成脂和成骨染色
图2 Nrf2-HUCMSCs成脂和成骨染色
图3 四组小鼠ALT、AST水平
图4 四组小鼠肝组织损伤情况
图5 四组小鼠肝细胞凋亡情况
[1]
WenBo W, Fei Z, YiHeng D, et al. Human umbilical cord mesenchymal stem cells overexpressing nerve growth factor ameliorate diabetic cystopathy in rats[J]. Neurochem Res, 2017, 42(12):3537-3547.
[2]
Zhang T, Wang P, Liu Y, et al. Overexpression of FOXQ1 enhances anti-senescence and migration effects of human umbilical cord mesenchymal stem cells in vitro and in vivo[J]. Cell Tissue Res, 2018, 373(2):379-393.
[3]
Rowart P, Erpicum P, Detry O, et al. Mesenchymal stromal cell therapy in ischemia/reperfusion injury[J]. J Immunol Res, 2015: 602597.
[4]
Ichihara S, Yamada Y, Liu F, et al. Ablation of the transcription factor Nrf2 promotes ischemia-induced neovascularization by enhancing the in flammatory response[J]. Arterioscler Thromb Vasc Biol, 2010, 30(8):1553-1561.
[5]
Morito N, Yoh K, Itoh K, et al. Nrf2 regulates the sensitivity of death receptor signals by affecting intracellular glutathione levels[J]. Oncogene, 2003, 22(58):9275-9281.
[6]
Kode A, Rajendrasozhan S, Caito S, et al. Resveratrol induces glutathione synthesis by activation of Nrf2 and protects against cigarette smoke-mediated oxidative stress in human lung epithelial cells[J]. Am J Physiol Lung Cell Mol Physiol, 2008, 294(3):L478-488.
[7]
Zhang S, Jiang W, Ma L, et al. Nrf2 transfection enhances the efficacy of human amniotic mesenchymal stem cells to repair lung injury induced by lipopolysaccharide[J]. J Cell Biochem, 2018, 119(2): 1627-1636.
[8]
Mohammadzadeh-Vardin M, Habibi Roudkenar M, Jahanian-Najafabadi A. Adenovirus-mediated over-expression of Nrf2 within mesenchymal stem cells (MSCs) protected rats against acute kidney injury[J]. Adv Pharm Bull, 2015, 5(2):201-208.
[9]
刘荣强，袁泽南，吴小材，等．核因子E2相关因子2过表达对间充质干细胞抗缺氧和抗凋亡能力的影响[J]．器官移植，2018, 9(2): 110-115.
[10]
Khan RS, Newsome PN. Non-alcoholic fatty liver disease and liver transplantation[J]. Metabolism, 2016, 65(8):1208-1223.
[11]
Nativ NI, Maguire TJ, Yarmush G, et al. Liver defatting: an alternative approach to enable steatotic liver transplantation[J]. Am J Transplant, 2012, 12(12):3176-3183.
[12]
Tashiro H, Kuroda S, Mikuriya Y, et al. Ischemia-reperfusion injury in patients with fatty liver and the clinical impact of steatotic liver on hepatic surgery[J]. Surg Today, 2014, 44(9):1611-1625.
[13]
Shende P, Subedi M. Pathophysiology, mechanisms and applications of mesenchymal stem cells for the treatment of spinal cord injury[J]. Biomed Pharmacother, 2017, 91:693-706.
[14]
Li Y, Hu G, Cheng Q. Implantation of human umbilical cord mesenchymal stem cells for ischemic stroke: perspectives and challenges[J]. Front Med, 2015, 9(1):20-29.
[15]
Roura S, Gálvez-Montón C, Bayes-Genis A. Umbilical cord blood-derived mesenchymal stem cells: new therapeutic weapons for idiopathic dilated cardiomyopathy?[J]. Int J Cardiol, 2014, 177(3): 809-818.
[16]
Fang X, Liu L, Dong J, et al. A study about immunomodulatory effect and efficacy and prognosis of human umbilical cord mesenchymal stem cells in patients with chronic hepatitis B-induced decompensated liver cirrhosis[J]. J Gastroenterol Hepatol, 2018, 33(4):774-780.
[17]
Sun Y, Wang Y, Zhou L, et al. Spheroid-cultured human umbilical cord-derived mesenchymal stem cells attenuate hepatic ischemia-reperfusion injury in rats[J]. Sci Rep, 2018, 8(1):2518.
[18]
Moshrefi M, Yari N, Nabipour F, et al. Transplantation of differentiated umbilical cord mesenchymal cells under kidney capsule for control of type I diabetes in rat[J]. Tissue Cell, 2015, 47(4):395-405.
[19]
Fahmy SR, Soliman AM, El Ansary M, et al. Therapeutic efficacy of human umbilical cord mesenchymal stem cells transplantation against renal ischemia/reperfusion injury in rats[J]. Tissue Cell, 2017, 49(3): 369-375.
[20]
Shimada S, Fukai M, Wakayama K, et al. Hydrogen sulfide augments survival signals in warm ischemia and reperfusion of the mouse liver[J]. Surg Today, 2015, 45(7):892-903.
[21]
Morales P, Vargas R, Videla LA, et al. Nrf2 activation in the liver of rats subjected to a preconditioning sub-chronic iron protocol[J]. Food Funct, 2014, 5(2):243-250.
[1] 孔莹莹, 谢璐涛, 卢晓驰, 徐杰丰, 周光居, 张茂. 丁酸钠对猪心脏骤停复苏后心脑损伤的保护作用及机制研究[J]. 中华危重症医学杂志(电子版), 2023, 16(05): 355-362.
[2] 张晓燕, 肖东琼, 高沪, 陈琳, 唐发娟, 李熙鸿. 转录因子12过表达对脓毒症相关性脑病大鼠大脑皮质的保护作用及其机制[J]. 中华妇幼临床医学杂志(电子版), 2023, 19(05): 540-549.
[3] 刘硕儒, 王功炜, 张斌, 李书豪, 胡成. 新型溶瘤病毒M1激活内质网应激致前列腺癌细胞凋亡的机制[J]. 中华腔镜泌尿外科杂志(电子版), 2023, 17(04): 388-393.
[4] 唐英俊, 李华娟, 王赛妮, 徐旺, 刘峰, 李羲, 郝新宝, 黄华萍. 人脐带间充质干细胞治疗COPD小鼠及机制分析[J]. 中华肺部疾病杂志(电子版), 2023, 16(04): 476-480.
[5] 李晔, 何洁, 胡锦秀, 王金祥, 田川, 潘杭, 陈梦蝶, 赵晓娟, 叶丽, 张敏, 潘兴华. 高活性间充质干细胞干预猕猴卵巢衰老的研究[J]. 中华细胞与干细胞杂志(电子版), 2023, 13(04): 210-219.
[6] 龙慧玲, 林蜜, 邵婷. 三维球体间充质干细胞培养技术的研究进展及其应用[J]. 中华细胞与干细胞杂志(电子版), 2023, 13(04): 229-234.
[7] 刘文慧, 吴涛, 张曦. 间充质干细胞联合血小板生成素受体激动剂在异基因造血干细胞移植后血小板恢复中的研究进展[J]. 中华细胞与干细胞杂志(电子版), 2023, 13(04): 242-246.
[8] 王红敏, 谢云波, 王彦虎, 王福生. 间充质干细胞治疗新冠病毒感染的临床研究进展[J]. 中华细胞与干细胞杂志(电子版), 2023, 13(04): 247-256.
[9] 李青霖, 宋仁杰, 周飞虎. 一种重型劳力性热射病相关急性肾损伤小鼠模型的建立与探讨[J]. 中华肾病研究电子杂志, 2023, 12(05): 265-270.
[10] 杨蕴钊, 周诚, 石美涵, 赵静, 白雪源. 人羊水间充质干细胞对膜性肾病大鼠的治疗作用[J]. 中华肾病研究电子杂志, 2023, 12(04): 181-186.
[11] 朱泽超, 杨新宇, 李侑埕, 潘鹏宇, 梁国标. 染料木黄酮通过SIRT1/p53信号通路对蛛网膜下腔出血后早期脑损伤的作用[J]. 中华神经创伤外科电子杂志, 2023, 09(05): 261-269.
[12] 于迪, 于海波, 吴焕成, 李玉明, 苏彬, 陈馨. 发状分裂相关增强子1差异表达对胆固醇刺激下血管内皮细胞的影响[J]. 中华脑科疾病与康复杂志(电子版), 2023, 13(05): 264-270.
[13] 王淑友, 宋晓晶, 贾术永, 王广军, 张维波. 肝脏去唾液酸糖蛋白受体靶向活体荧光成像评估酒精性肝损伤肝脏功能的研究[J]. 中华消化病与影像杂志(电子版), 2023, 13(06): 443-446.
[14] 梁宇同, 丁旭, 马国慧, 黄艳红. 间充质干细胞在宫腔粘连治疗中的研究进展[J]. 中华临床医师杂志(电子版), 2023, 17(05): 596-599.
[15] 邱甜, 杨苗娟, 胡波, 郭毅, 何奕涛. 亚低温治疗脑梗死机制的研究进展[J]. 中华脑血管病杂志(电子版), 2023, 17(05): 518-521.
阅读次数
全文


摘要

版权所有 中华医学会 中华医学电子音像出版社有限责任公司  京ICP备14006079号-1  网络出版服务许可证：(署)网出证(京)字第075号