[1]高帅,夏安周,侯筱宇,等. 肾缺血辕再灌注损伤中近曲小管Legumain表达的研究[J].徐州医科大学学报,2017,37(05):281-286.
 GAO Shuai,XIA Anzhou,HOU Xiaoyu,et al. Expression of legumain in the proximal tubule after renal ischemia/reperfusion injury[J].Journal of Xuzhou Medical University,2017,37(05):281-286.





 Expression of legumain in the proximal tubule after renal ischemia/reperfusion injury
 (上海交通大学附属第一人民医院泌尿外科,上海;徐州医科大学药理学教研室,江苏徐州; 徐州医科大学基础医学院生物化学与分子生物学研究中心
 GAO Shuai1 XIA Anzhou2 HOU Xiaoyu3 GUO Yifeng1*
 1.Department of Urology, Shanghai General Hospital, Shanghai Jiao Tong University, Shanghai 200080, China;
2.Department of Pharmacology, Xuzhou Medical University, Xuzhou, Jiangsu 221004;3.Research Center for
Biochemistry and Molecular Biology, College of Biomedical Science, Xuzhou Medical University
 kidneys ischemia/reperfusion injury legumain
目的 建立小鼠肾缺血/再灌注损伤模型,观察小鼠肾缺血/再灌注后的损伤、Legumain的表达,探讨Legumain在小鼠肾脏缺血/再灌注损伤中的作用及可能机制。方法 30只C57BL/6小鼠行右肾切除后随机分为缺血/再灌注损伤组(IRI组)、对照组,夹闭左肾蒂45 min缺血,缺血后再灌注不同时间(1 h、6 h、24 h),检测各组小鼠血肌酐、尿素氮水平,观察肾组织形态学变化,免疫组化技术与蛋白印迹分析Legumain的表达。结果 IRI组肾脏经过缺血/再灌注后,血肌酐及尿素氮水平在1 h、6 h、24 h再灌注时间点较对照组明显升高(P<0.05)。IRI组小鼠肾损伤程度较对照组明显加重,表现为肾小管细胞坏死脱落、间质水肿、炎症细胞浸润增多,损伤随着再灌注时间的延长而加重。IRI组肾近曲小管中Legumain的表达较对照组明显增强(P<0.01)。结论 肾缺血后再灌注会造成损伤,导致肾功能下降、肾小管细胞损伤坏死、Legumain表达增强,Legumain的表达变化可能在肾缺血/再灌注损伤中起重要作用。
 Objective To establish a mouse model of renal ischemia/reperfusion injury, and investigate the renal ischemia/reperfusion injury, the expression and role of legumain in mice with renal ischemia/reperfusion injury and possible mechanisms. Methods A total of 30 C57BL/6 mice were randomly divided into two groups after right renal resection: an ischemia/reperfusion injury (IRI) group and a control group. The mice were subject to blockage of the left renal pedicle for 45 min followed by reperfusion for various times (1, 6 and 24 h). Then, the levels of serum creatinine and urea nitrogen and morphological changes of each group were observed, while the expression of legumain was examined by immunohistochemistry and western blotting. Results After renal ischemia/reperfusion, the IRI group produced remarkably higher levels of serum creatinine and urea nitrogen than the control group after reperfusion for 1, 6, and 24 h (P<0.05). Compared with the control group, the IRI group presented markedly worsened renal injury which was characterized in necrosis of the renal tubular cells, interstitial edema, and increased infiltration of inflammatory cells, and aggravated as reperfusion extended. The expression of legumainin in the proximal tubule of the IRI group was significantly higher than that in the control group (P<0.01). Conclusions Renal ischemia/reperfusion can lead to decreased renal function, renal tubular damage and higher expression of legumain. The change in the expression of legumain may play an important role in ischemia/reperfusion injury.


 [1] Bonventre JV, Yang L. Cellular pathophysiology of ischemic acute kidney injury [J]. J Clin Invest, 2011,121(11):4210-4221.
[2] Sabbahy ME, Vaidya VS. Ischemic kidney injury and mechanisms of tissue repair [J]. Wiley Interdiscip Rev SystBiol Med, 2010,3(5):606-618.
[3] Sun Y, Fujigaki Y, Sakakima M, et al. Acquired resistance to rechallenge injury in rats recovered from subclinical renal damage with uranyl acetate-Importance of proliferative activity of tubular cells [J]. Toxicol Appl Pharmacol, 2010,243(1):104-110.
[4] Takaori K, Nakamura J, Yamamoto S, et al. Severity and frequency of proximal tubule injury determines renal prognosis [J]. J Am Soc Nephrol, 2015,27(8):2393-2399.
[5] Ma D, Lim T, Xu J, et al. Xenon preconditioning protects against renal ischemic-reperfusion Injury via HIF-1α activation [J]. J Am Soc Nephrol, 2009,20(4):713-720.
[6] 陈越,蒋素华,朱加明,等.小分子RNA干扰沉默缺氧诱导因子1α加重缺氧状态肾小管上皮细胞的生长抑制和坏死[J]. 中华肾脏病杂志,2010,26(7):530-536.
[7] Mizuno S, Fujita K, Furuy R, et al. Association of HSP73 with the acquired resistance to uranyl acetate-induced acute renal failure [J]. Toxicology, 1997,117(2-3):183-191.
[8] Fujikura T, Togawa A, Sun Y, et al. Dephosphorylated Ser985 of c-Met is associated with acquired resistance to rechallenge injury in rats that had recovered from uranyl acetate-induced subclinical renal damage [J]. Clin Exp Nephrol, 2013,17(4):504-514.
[9] Chen JM, Rawlings ND, Stevens RA, et al. Identification of the active site of legumain links it to caspases, clostripain and gingipains in a new clan of cysteine endopeptidases [J]. FEBS Lett, 1998,441(3):361-365.
[10] Chen JM, Dando PM, Rawlings ND, et al. Cloning, isolation, and characterization of mammalian legumain, an asparaginyl endopeptidase [J]. J Biol Chem, 1997,272(12):8090-8098.
[11] Chen JM, Dando PM, Stevens RA, et al. Cloning and expression of mouse legumain, a lysosomal endopeptidase [J]. Biochem J, 1998,335(Pt 1):111-117.
[12] Miller G, Matthews SP, Reinheckel T, et al. Asparagine endopeptidase is required for normal kidney physiology and homeostasis [J]. FASEB J, 2011,25(5):1606-1617.
[13] Morita Y, Araki H, Sugimoto T, et al. Legumain/asparaginyl endopeptidase controls extracellular matrix remodeling through the degradation of fibronectin in mouse renal proximal tubular cells [J]. FEBS Lett, 2007,581(7):1417-1424.
[14] Watts C, Matthews SP, Mazzeo D, et al. Asparaginyl endopeptidase: case history of a class II MHC compartment protease [J]. Immunol Rev, 2005,207(1):218-228.
[15] Mattock KL, Gough PJ, Humphries J, et al. Legumain and cathepsin-L expression in human unstable carotid plaque [J]. Atherosclerosis, 2010,208(1):83-89.
[16] Wu M, Shao GR, Zhang FX, et al. Legumain protein as a potential predictive biomarker for Asian patients with breast carcinoma [J]. Asian Pac J Cancer Prev, 2014,15(24):10773-10777.
[17] Haugen MH, Boye K, Nesland JM, et al. High expression of the cysteine proteinase legumain in colorectal cancer - implications for therapeutic targeting [J]. Eur J Cancer, 2015,51(1):9-17.
[18] Li N, Liu Q, Su Q, et al. Effects of legumain as a potential prognostic factor on gastric cancers[J/OL]. Med Oncol, 2013,30(3):621. doi: 0.1007/s12032-013-0621-9.
[19] Murthy RV, Arbman G, Gao J, et al. Legumain expression in relation to clinicopathologic and biological variables in colorectal cancer [J]. Clin Cancer Res, 2009,11(6):2293-2299.
[20] Wang L, Chen S, Zhang M, et al. Legumain: a biomarker for diagnosis and prognosis of human ovarian cancer [J]. J Cell Biochem, 2012,113(8):2679-2686.
[21] Zhen Y, Guo C, Shen W, et al. Clinicopathologic significance of legumain overexpression in cancer: a systematic review and meta-analysis [J/OL]. Sci Rep, 2015,5:16599. doi: 10.1038/srep16599.
[22] 吴文苑,余涟,刘春,等.Legumain对肿瘤细胞侵润力和血管内皮细胞管腔形成的作用[J].热带医学杂志,2009,9(8):864-867.
[23] 胡红林,王共先,邹丛,等.BALB/c小鼠肾缺血再灌注损伤模型的建立与评价[J].中国组织工程研究,2011,15(5):870-873.
[24] Furuichi K, Wada T, Iwata Y, et al. Administration of FR167653, a new anti-inflammatory compound, prevents renal ischaemia/reperfusion injury in mice [J]. Nephrol Dial Transplant, 2002,17(3):399-407.
[25] Dall E, Brandstetter H. Structure and function of legumain in health and disease [J]. Biochimie, 2016,122:126-150.
[26] Wang J, Huo K, Ma L, et al. Toward an understanding of the protein interaction network of the human liver [J]. Mol Syst Biol, 2011,7(1):536-536.
[27] Thorsteinsdóttir S, Roelen BA, Freund E, et al. Expression patterns of laminin receptor splice variants α6Aβ1 and α6Bβ1 suggest different roles in mouse development [J]. Dev Dyn, 1995,204(3):240-258.
[28] Giancotti FG, Ruoslahti E. Integrin signaling [J]. Science, 1999,285(5430):1028-1032.
[29] Mccubrey JA, Steelman LS, Franklin RA, et al. Targeting the RAF/MEK/ERK, PI3K/AKT and P53 pathways in hematopoietic drug resistance [J]. Adv Enzyme Regul, 2007,47(1):64-103.
[30] Arcaro A, Guerreiro AS. The phosphoinositide 3-kinase pathway in human cancer: genetic alterations and therapeutic implications [J]. Curr Genomics, 2007,8(5):271-306.
[31] Vandermoere F, Yazidibelkoura IE, Adriaenssens E, et al. The antiapoptotic effect of fibroblast growth factor-2 is mediated through nuclear factor-κB activation induced via interaction between Akt and IκB kinase-β in breast cancer cells [J]. Oncogene, 2005,24(35):5482-5491.
[32] Jeong SJ, Pisemasison CA, Radonovich MF, et al. Activated AKT regulates NF-κB activation, p53 inhibition and cell survival in HTLV-1-transformed cells [J]. Oncogene, 2005,24(44):6719-6728.


[2]代春潇,宋远见,张云山,等. 大鼠MKK7真核表达质粒的构建及其在COS7细胞中转染效率[J].徐州医科大学学报,2017,37(04):211.
  [J].Journal of Xuzhou Medical University,2017,37(05):211.
[3]张端强,丁晓帆,杨丹丹,等. 加综合征患者复发影响因素研究[J].徐州医科大学学报,2017,37(04):216.
 [J].Journal of Xuzhou Medical University,2017,37(05):216.
[4]王立新,袁峰*,郭开今,等. 纳米羟基磷灰石/细菌纤维素对兔骨髓间充质干细胞黏附及成骨分化的影响[J].徐州医科大学学报,2017,37(04):221.
 [J].Journal of Xuzhou Medical University,2017,37(05):221.
[5]李海青,曹猛,江涛,等. PD-L1在结直肠癌中的表达及其临床意义[J].徐州医科大学学报,2017,37(04):227.
 [J].Journal of Xuzhou Medical University,2017,37(05):227.
[6]刘兵,唐明生,李苹,等. 用随机森林模型分析胃癌高危人群干预效果的影响因素*[J].徐州医科大学学报,2017,37(04):231.
 [J].Journal of Xuzhou Medical University,2017,37(05):231.
[7]周新宇?,牛坚,曹宽,等. Tim3在炎症反应调控及对肝细胞损伤保护机制研究[J].徐州医科大学学报,2017,37(04):236.
 [J].Journal of Xuzhou Medical University,2017,37(05):236.
[8]商书霞,宋光耀*,刘晓耕,等. 血清胆红素和γ-谷氨酰转肽酶与糖尿病患者非酒精性脂肪肝的相关性研究[J].徐州医科大学学报,2017,37(04):245.
 [J].Journal of Xuzhou Medical University,2017,37(05):245.
[9]李新文1,巩尊科2,王世雁2,等. RS在非痴呆型血管性认知障碍评定中的临床应用研究[J].徐州医科大学学报,2017,37(04):248.
 [J].Journal of Xuzhou Medical University,2017,37(05):248.
 [J].Journal of Xuzhou Medical University,2017,37(05):252.


更新日期/Last Update: 2017-06-09