[1]杨东海,韩愈,巩正藩,等.G蛋白偶联受体激酶4调控小鼠肾脏急性缺血再灌注损伤的机制探讨[J].第三军医大学学报,2018,40(08):666-672.
 YANG Donghai,HAN Yu,GONG Zhengfan,et al.Role of G protein-coupled receptor kinase 4 in regulation of acute renal ischemia-reperfusion injury in mice[J].J Third Mil Med Univ,2018,40(08):666-672.
点击复制

G蛋白偶联受体激酶4调控小鼠肾脏急性缺血再灌注损伤的机制探讨(/HTML )
分享到:

《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
40卷
期数:
2018年第08期
页码:
666-672
栏目:
基础医学
出版日期:
2018-04-30

文章信息/Info

Title:
Role of G protein-coupled receptor kinase 4 in regulation of acute renal ischemia-reperfusion injury in mice
作者:
杨东海韩愈巩正藩周中淑吴连判傅春江曾春雨周林
陆军军医大学(第三军医大学)第三附属医院野战外科研究所心血管内科,重庆市心血管病研究所
Author(s):
YANG Donghai HAN Yu GONG Zhengfan ZHOU Zhongshu WU Lianpan FU Chunjiang ZENG Chunyu ZHOU Lin

Department of Cardiology, Chongqing Institute of Cardiology,  Institute of Surgery Research, Third Affiliated Hospital, Army Medical University (Third Military Medical University), Chongqing, 400042, China

关键词:
G蛋白偶联受体激酶4肾脏缺血再灌注损伤血管紧张素Ⅱ1型受体凋亡氧化应激
Keywords:
G protein-coupled receptor kinase 4 renal ischemia-reperfusion injury angiotensin Ⅱ receptor 1 apoptosis oxidative stress
分类号:
R364.12;R692;R977.3
文献标志码:
A
摘要:

目的    探讨G蛋白偶联受体激酶4(G proteincoupled receptor kinase 4,GRK4)对小鼠肾脏急性缺血再灌注损伤的影响及其作用机制。方法    取SPF级健康野生型[8周龄、体质量(21.34±042)g]和GRK4转基因型[8周龄、体质量(21.87±0.68)g] C57BL/6小鼠,各12只。各型分别按随机数字表法分为4组(n=6):野生型假手术对照组、野生型肾脏缺血再灌注损伤组、GRK4转基因型假手术对照组、GRK4转基因型肾脏缺血再灌注损伤组。假手术对照组均采用开腹后不阻断肾动脉血流;缺血再灌注损伤组均采用夹闭肾动脉缺血45 min再灌注24 h,建立小鼠肾脏I/R模型。各组处死小鼠后,取血标本进行肾功能检测(血肌酐、血尿素氮);HE染色观察肾脏病理形态改变,并行肾小管损伤半定量评分;测定肾脏组织中过氧化物歧化酶(superoxide dismutase,SOD)、丙二醛(malondialdehyde,MDA)等氧化应激水平改变;TUNEL染色检测肾脏组织中细胞凋亡情况;蛋白质免疫印记方法检测各组小鼠肾脏组织中GRK4和AT1受体的蛋白表达变化。结果    野生型小鼠肾脏I/R模型后肾功能受损,血肌酐、血尿素氮升高,肾脏小管上皮细胞脱落、死亡(P<0.05);肾脏组织中GRK4蛋白表达含量增加,差异有统计学意义(P<0.05)。在GRK4过表达小鼠上研究结果发现,过表达GRK4的肾脏在缺血再灌注损伤后,肾功能损害进一步加重;肾脏病理损伤评分明显增加(P<0.05)。肾脏氧化应激水平明显上升,总SOD下降和MDA升高(P<0.05);肾脏凋亡细胞数目显著增多(P<0.05)。肾脏组织中AT1受体表达量增加(P<0.05),AT1受体含量的升高可以加重小管细胞氧化应激和凋亡的发生。结论    GRK4可以通过上调肾脏AT1受体表达,增加肾脏氧化应激水平和肾小管细胞凋亡,加重肾脏缺血再灌注损伤。

Abstract:

Objective     To investigate the regulatory role of G protein-coupled receptor kinase 4 (GRK4) in acute renal ischemia-reperfusion (I/R) injury in mice and explore its mechanism. Methods    Twelve 8-week-old wild-ype C57BL/6 mice (body weight 21.34±0.42 g) and 12 age-matched GRK4 transgenic C57BL/6 mice (body weight 21.87±0.68 g) were both randomized into sham-operated group and renal I/R group (n=6). In the 2 I/R groups, the mice underwent laparotomy and the renal artery ischemia was clipped for 45 min followed by reperfusion for 24 h; For sham operation, the renal artery was exposed without occlusion. After the operation, blood samples were collected to measure serum creatinine and blood urea nitrogen levels. The pathological changes of the kidney were observed with HE staining and graded, and TUNEL staining was used to detect cell apoptosis in the renal tissue. Superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in the renal tissue were measured to evaluate the changes of oxidative stress. Renal expressions of GRK4 and AT1R were measured with Western blotting. Results    The wild-type mice had significantly increased serum creatinine and blood urea nitrogen levels after renal I/R injury with obvious renal pathologies shown by renal tubular epithelial cell loss and death (P<0.05); the expression of GRK4 in the kidneys was significantly increased after renal I/R injury (P<0.05). Compared with wildtype mice, GRK4 transgenic mice exhibited significant impairment of renal function after I/R injury with worsened renal pathologies, significantly increased renal cell apoptosis (P<0.05), lowered SOD activity (P<0.05), increased MDA content (P<0.05), and up-regulated expression of AT1R in the renal tissue (P<0.05). Conclusion    GRK4 aggravates acute renal I/R injury and increases apoptosis and oxidative stress in renal tubular cells by upregulating AT1R expression.

参考文献/References:

[1]BELLOMO R, KELLUM J A, RONCO C. Acute kidney injury:The Lancet[J]. Lancet, 2012, 380(9843): 756-766. DOI: 10.1016/S01406736(11)61454-61462.
[2]FORTRIE G, STADS S, AARNOUDSE A J, et al. Longterm sequelae of severe acute kidney injury in the critically ill patient without comorbidity: a retrospective cohort study[J]. PLoS One, 2015, 10(3): 121482-121494. DOI: 10.1371/journal.pone.0121482.
[3]MACEDO E, MEHTA R L. Renal recovery after acute kidney injury[J]. Contrib Nephrol, 2016, 187: 24-35. DOI: 10.1159/000443289.
[4]YATABE J, SANADA H, MIDORIKAWA S, et al. Effects of decreased renal cortical expression of G protein-coupled receptor kinase 4 and angiotensin type 1 receptors in rats[J]. Hypertens Res, 2008, 31(7): 1455-1464. DOI: 10.1291/hypres.31.1455.
[5]JOSE P A, SOARESDASILVA P, EISNER G M, et al. Dopamine and G proteincoupled receptor kinase 4 in the kidney: role in blood pressure regulation[J]. Biochim Biophys Acta, 2010, 1802(12): 1259-1267. DOI: 10.1016/j.bbadis.2010.02.004.
[6]YANG J, VILLAR V A, JONES J E, et al. G Protein-Coupled Receptor Kinase 4: Role in Hypertension[J]. Hypertension, 2015, 65(6): 1148-1155. DOI: 10.1161/HYPERTENSIONAHA.115.05189.
[7]HUMPHREYS B D, VALERIUS M T, KOBAYASHI A, et al. Intrinsic epithelial cells repair the kidney after injury [J]. Cell Stem Cell, 2008, 2(3): 284. DOI: 10.1016/j.stem.2008.01.014.
[8]王永斌, 韩愈, 高照, 等. 白藜芦醇通过沉默信息调节因子1保护造影剂急性肾损伤[J]. 中华高血压杂志, 2016(12): 1154-1160. DOI: 10.16439/j.cnki.16737245.2016.12.017.
WNAG Y B, HAN Y, GAO Z, et al. Resveratrol protects against contrast-induced nephropathy in rats through silent information regulator1 [J]. Chin J Hypertens, 2016(12): 1154-1160. DOI: 10.16439/j.cnki.1673-7245.2016.12.017.
[9]WANG Z, ZENG C, VILLAR V A, et al. Human GRK4γ142V variant promotes angiotensin II type I receptor-mediated hypertension via renal histone deacetylase type 1 inhibition [J]. Hypertension, 2016, 67(2): 325-334. DOI: 10.1161/HYPERTENSIONAHA.115.05962.
[10]LINGYONG L, HOMAN K T, VISHNIVETSKIY S A, et al. G proteincoupled receptor kinases of the GRK4 protein subfamily phosphorylate inactive G proteincoupled receptors (GPCRs)[J]. J Biol Chem. 2015, 290(17): 10775-10790. DOI: 10.1074/jbc.M115.644773.
[11]POLHEMUS D J, GAO J, SCARBOROUGH A L, et al. Radiofrequency renal denervation protects the ischemic heart via inhibition of GRK2 and increased nitric oxide signaling [J]. Circ Res, 2016, 119(3): 470-480. DOI: 10.1161/CIRCRESAHA.115.308278.
[12]BAEK J H, ZENG R, WEINMANNMENKE J, et al. IL-34 mediates acute kidney injury and worsens subsequent chronic kidney disease[J]. J Clin Invest, 2015, 125(8): 3198-3214. DOI: 10.1172/JCI81166.
[13]ELTZSCHIG H K, ECKLE T. Ischemia and reperfusion-from mechanism to translation[J]. Nat Med, 2011 Nov 7;17(11): 1391-1401. DOI: 10.1038/nm.2507.
[14]XU Y, MA H, SHAO J, et al. A role for tubular necroptosis in cisplatin-induced AKI[J]. J Am Soc Nephrol, 2015, 26(11): 2647-658. DOI: 10.1681/ASN.2014080741.
[15]CAO W, LI A, LI J, et al. Reno-cerebral reflex activates the reninangiotensin system, promoting oxidative stress and renal damage after ischemia-reperfusion injury [J]. Antioxid Redox Signal, 2017, 27(7): 415-432. DOI: 10.1089/ars.2016.6827.
[16]FANG F, LIU G C, ZHOU X, et al. Loss of ACE2 exacerbates murine renal ischemiareperfusion injury [J]. PLoS One, 2013, 8(8): e71433. DOI: 10.1371/journal.pone.0071433.
[17]CAO W, LI A, LI J, et al. Reno-cerebral reflex activates the reninangiotensin system, promoting oxidative stress and renal damage after ischemia-reperfusion injury[J]. Antioxid Redox Signal, 2017, 27(7): 415-432. DOI: 10.1089/ars.2016.6827.
[18]WANG Z, LIU Y, HAN Y, et al. Protective effects of aliskiren on ischaemia-reperfusion-induced renal injury in rats[J]. Eur J Pharmacol, 2013, 718(1-3): 160-166. DOI: 10.1016/j.ejphar.2013.08.038.

更新日期/Last Update: 2018-04-27