网站首页期刊介绍通知公告编 委 会投稿须知电子期刊广告合作联系我们
最新消息:
LRP-1抑制缺氧/复氧诱导的肾细胞损伤
作者:段斌1  职瑾2  杜鹏1  高妍婷1  李振江1 
单位:1. 陕西省人民医院 肾病血透中心, 陕西 西安 710068;
2. 西安市第一医院, 陕西 西安 710002
关键词:低密度脂蛋白受体相关蛋白1 缺氧/复氧 肾细胞 JNK 
分类号:R33-33
出版年·卷·期(页码):2017·36·第十二期(1697-1704)
摘要:

目的:探讨低密度脂蛋白受体相关蛋白1(LRP-1)在肾小管上皮细胞(HK2)缺氧/复氧损伤中的作用及其可能机制。方法:建立HK-2缺氧/复氧模型;实时荧光定量PCR (qRT-PCR)检测LRP-1 mRNA表达量;Western blotting检测LRP-1、JNK蛋白表达量和JNK磷酸化水平;构建重组载体pcDNA.3.1-LRP-1并转染细胞过表达LRP-1;转染LRP-1 siRNA抑制LRP-1表达;试剂盒方法检测caspase-3活性;Annexin-Ⅴ fluorescein isothiocyanate conjugate and propidium iodide (Annexin-Ⅴ FITC/PI)方法检测细胞凋亡;MTT法检测细胞生长活力;试剂盒方法检测乳酸脱氢酶(LDH)漏出量;JNK抑制剂SP600125孵育HK-2。结果:HK-2缺氧/复氧诱导LRP-1表达量上调;过表达LRP-1显著下调JNK磷酸化水平,降低缺氧/复氧诱导的HK-2凋亡,降低caspase-3活性和LDH漏出量,且提高细胞生长活力。抑制LRP-1则作用相反。另外,SP600125抑制JNK磷酸化水平,且提高细胞生长活力,降低caspase-3活性。结论:LRP-1可通过调控JNK降低肾细胞的缺氧/复氧损伤,为肾脏缺血再灌注的防治提供新的靶点。

Objective: To explore the effects of low density lipoprotein receptor-related protein 1(LRP-1) on hypoxia/reoxygenation-induced renal tubular epithelial cell (HK-2) injury and the related mechanism.Methods: The model of HK-2 with hypoxia/reoxygenation injury was established. The mRNA expression of LRP-1 was measured by qRT-PCR. The protein expressions of LRP-1 and JNK and JNK phosphorylation level were detected using Western blotting. Recombinant vector pcDNA.3.1-LRP-1 was constructed and transfected into cells to overexpress LRP-1. LRP-1 siRNA was transfected into cells to inhibit the expression of LRP-1. Caspase-3 activity was detected by using the kit. Cell apoptosis was tested by Annexin-Ⅴ fluorescein isothiocyanate conjugate and propidium iodide (Annexin-Ⅴ FITC/PI) assay. Cell viability was detected by MTT. LDH leakage was quantified by using the kit. HK2 was incubated with JNK inhibitor SP600125.Results: LRP-1 was induced by the hypoxia/reoxygenation injury in HK-2. LRP-1 overexpression obviously down-regulated JNK phosphorylation level, hypoxia/reoxygenation-induced cells apoptosis, caspase-3 activity and LDH leakage, whereas promoted cell viability. LRP-1 inhibition had the reversed effect. What's more, SP600125 decreasd the JNK phosphorylation level and LDH leakage, whereas increased cell viability.Conclusions: LRP-1 can constrain the kidney cells injury caused by hypoxia/reoxygenation via regulating JNK, providing the theoretical basis for the treatment of renal ischemia reperfusion.

参考文献:

[1] LORENZEN J M,KAUCSAR T,SCHAUERTE C,et al.MicroRNA-24 antagonism prevents renal ischemia reperfusion injury[J]. J Am Soc Nephrol,2014,25(12):2717-2729.
[2] SALAMEH A,DHEIN S.Strategies for Pharmacological Organoprotection during Extracorporeal Circulation Targeting Ischemia-Reperfusion Injury[J].Front Pharmacol,2015,6:296.
[3] KIM Y W,CHA K C,CHA Y S,et al.Shock duration after resuscitation is associated with occurrence of post-cardiac arrest acute kidney injury[J].J Korean Med Sci,2015,30(6):802-807.
[4] WUJAK L,MARKART P,WYGRECKA M.The low density lipoprotein receptor-related protein (LRP) 1 and its function in lung diseases[J].Histol Histopathol,2016,31(7):733-745.
[5] CANUEL M,SUN X,ASSELIN M C,et al.Proprotein convertase subtilisin/kexin type 9(PCSK9) can mediate degradation of the low density lipoprotein receptor-related protein 1(LRP-1)[J].PLoS One,2013,8(5):e64145.
[6] MAY P,BOCK H H,NOFER J R.Low density receptor-related protein 1(LRP1) promotes anti-inflammatory phenotype in murine macrophages[J].Cell Tissue Res,2013,354(3):887-889.
[7] XU H,PERREAU V M,DENT K A,et al.Iron regulates apolipoprotein E expression and secretion in neurons and astrocytes[J].J Alzheimers Dis,2016,51(2):471-487.
[8] LIN L,HU K.LRP-1:functions,signaling and implications in kidney and other diseases[J].Int J Mol Sci,2014,15(12):22887-22901.
[9] ZHAO Y,LI D,ZHAO J,et al.The role of the low-density lipoprotein receptor-related protein 1(LRP-1) in regulating blood-brain barrier integrity[J].Rev Neurosci,2016,27(6):623-634.
[10] LIN L,HU K.Tissue plasminogen activator:side effects and signaling[J]. J Drug Des Res,2014,1(1).pii:1001.
[11] WECKBACH L T,MURAMATSU T,WALZOG B.Midkine in inflammation[J].Scientific World Journal,2011,11:2491-2505.
[12] 王宇,刘菊英,曾文静,等.PTD4-Cu/Zn SOD融合蛋白对大鼠心肌细胞缺氧复氧损伤的影响[J].现代医学,2017,45(10):1396-1399.
[13] 承燕,吕磊,江时森.黄芪甲苷的抗氧化性对缺氧缺糖H9c2细胞的保护作用探讨[J].东南大学学报:医学版,2012,31(1):51-55.
[14] IBANEZ B,HEUSCH G,OVIZE M,et al.Evolving therapies for myocardial ischemia/reperfusion injury[J].J Am Coll Cardiol,2015,65(14):145414-145471.
[15] KALOGERIS T,BAO Y,KORTHUIS R J.Mitochondrial reactive oxygen species:a double edged sword in ischemia/reperfusion vs preconditioning[J].Redox Biol,2014,2:702-714.
[16] LINKERMANN A,BRASEN J H,DARDING M,et al.Two independent pathways of regulated necrosis mediate ischemia-reperfusion injury[J].Proc Natl Acad Sci U S A,2013,110(29):12024-12029.
[17] DARE A J,BOLTON E A,PETTIGREW G J,et al.Protection against renal ischemia-reperfusion injury in vivo by the mitochondria targeted antioxidant MitoQ[J].Redox Biol,2015,5:163-168.
[18] APPERT-COLLIN A,BENNASROUNE A,JEANNESSON P,et al.Role of LRP-1 in cancer cell migration in 3-dimensional collagen matrix[J]. Cell Adh Migr,2017,11(4):316-326.
[19] LIN L,HU K.Tissue plasminogen activator and inflammation:from phenotype to signaling mechanisms[J].Am J Clin Exp Immunol,2014,3(1):30-36.
[20] GONIAS S L,CAMPANA W M.LDL receptor-related protein-1:a regulator of inflammation in atherosclerosis,cancer,and injury to the nervous system[J].Am J Pathol,2014,184(1):18-27.
[21] KANG H S,KIM J,LEE H J,et al.LRP1-dependent pepsin clearance induced by 2'-hydroxycinnamaldehyde attenuates breast cancer cell invasion[J].Int J Biochem Cell Biol,2014,53:15-23.
[22] CHENG C F,FAN J,FEDESCO M,et al.Transforming growth factor alpha (TGFalpha)-stimulated secretion of HSP90alpha:using the receptor LRP-1/CD91 to promote human skin cell migration against a TGFbeta-rich environment during wound healing[J].Mol Cell Biol,2008,28(10):3344-3358.
[23] YANG E,ZHENG H,PENG H,et al.Lentivirus-induced knockdown of LRP1 induces osteoarthritic-like effects and increases susceptibility to apoptosis in chondrocytes via the nuclear factor-kappaB pathway[J].Exp Ther Med,2015,10(1):97-105.
[24] SAFINA D,SCHLITT F,ROMEO R,et al.Low-density lipoprotein receptor-related protein 1 is a novel modulator of radial glia stem cell proliferation,survival,and differentiation[J].Glia,2016,64(8):1363-1380.
[25] LOK K Z,MANZANERO S,ARUMUGAM T V.Neuronal low-density lipoprotein receptor-related protein 1(LRP1) enhances the anti-apoptotic effect of intravenous immunoglobulin (IVIg) in ischemic stroke[J].Brain Res,2016,1644:192-202.
[26] CAL R,JUAN-BABOT O,BROSSA V,et al.Low density lipoprotein receptor-related protein 1 expression correlates with cholesteryl ester accumulation in the myocardium of ischemic cardiomyopathy patients[J].J Transl Med,2012,10:160.
[27] SHIKUMA N J,ANTOSHECHKIN I,MEDEIROS J M,et al.Stepwise metamorphosis of the tubeworm Hydroides elegans is mediated by a bacterial inducer and MAPK signaling[J].Proc Natl Acad Sci U S A,2016,113(36):10097-10102.
[28] CLAVIER A,RINCHEVAL-ARNOLD A,BAILLET A,et al.Two different specific JNK activators are required to trigger apoptosis or compensatory proliferation in response to Rbf1 in Drosophila[J].Cell Cycle,2016,15(2):283-294.
[29] PUSHPAVALLI S N,SARKAR A,RAMAIAH M J,et al.Drosophila MOF regulates DIAP1 and induces apoptosis in a JNK dependent pathway[J].Apoptosis,2016,21(3):269-282.
[30] ZHAO M,HOWARD E W,PARRIS A B,et al.Alcohol promotes migration and invasion of triple-negative breast cancer cells through activation of p38 MAPK and JNK[J]. Mol Carcinog,2017,56(3):849-862.
[31] DHANASEKARAN D N,REDDY E P.JNK signaling in apoptosis[J].Oncogene,2008,27(48):6245-6251.
[32] SHANG L,ANANTHAKRISHNAN R,LI Q,et al.RAGE modulates hypoxia/reoxygenation injury in adult murine cardiomyocytes via JNK and GSK-3beta signaling pathways[J].PLoS One,2010,5(4):e10092.
[33] GARAY M,GAARDE W,MONIA B P,et al.Inhibition of hypoxia/reoxygenation-induced apoptosis by an antisense oligonucleotide targeted to JNK1 in human kidney cells[J].Biochem Pharmacol,2000,59(9):1033-1043.
[34] LANGLOIS B,PERROT G,SCHNEIDER C,et al.LRP-1 promotes cancer cell invasion by supporting ERK and inhibiting JNK signaling pathways[J].PLoS One,2010,5(7):e11584.

服务与反馈:
文章下载】【发表评论】【查看评论】【加入收藏
提示:您还未登录,请登录!点此登录
您是第 730134 位访问者


 ©《现代医学》编辑部
联系电话:025-83272481;83272479
电子邮件: xdyx@pub.seu.edu.cn

苏ICP备09058541