脓毒症中血小板活化对肾小管上皮影响的机制研究

doi:10.12092/j.issn.1009-2501.2018.03.007

中国临床药理学与治疗学 ›› 2018, Vol. 23 ›› Issue (3): 283-289.doi: 10.12092/j.issn.1009-2501.2018.03.007

脓毒症中血小板活化对肾小管上皮影响的机制研究

方晶晶，陶静，黄钦，史超路，颜碧清，盖磊，厉旭光

宁波大学医学院附属医院ICU，宁波 315020，浙江

收稿日期:2017-11-10 修回日期:2018-02-28 出版日期:2018-03-26 发布日期:2018-03-28
通讯作者: 颜碧清，男，本科，主任医师，研究方向：重症感染、脓毒症休克、多脏器功能衰竭、机械通气、镇静镇痛、血液净化、危重症营养支持等。
作者简介:方晶晶，女，研究生，主治医师，研究方向：脓毒症休克、机械通气、镇静镇痛、血液净化、危重症营养支持等。
基金资助:
C50-宁波市社会科学发展项目（201301C5010115）

Effects of platelet activation on renal tubular epithelial cells in sepsis

FANG Jingjing, TAO Jing, HUANG Qin, SHI Chaolu, YAN Biqing, GAI Lei, LI Xuguang

Intensive Care Unit，the Affiliated Hospital of Medical College of Ningbo University, Ningbo 315020, Zhejiang, China

Received:2017-11-10 Revised:2018-02-28 Online:2018-03-26 Published:2018-03-28

摘要/Abstract

摘要：

目的:探讨脓毒症中血小板活化对肾小管上皮影响的机制。方法: ELISA法检测血小板分泌的炎症因子白介素（IL）-1β和IL-6，应用共培养体系将血小板和肾小管上皮细胞共培养，Western blot检测下室肾小管上皮细胞的p65和p-p65蛋白，qRT-PCR方法检测肾小管上皮细胞中IL-1β和IL-6的mRNA表达水平。构建盲肠结扎穿孔的脓毒症模型，尾静脉注射TLR4特异性抑制剂TAK-242，EdU检测肾小管增殖情况，肾组织石蜡包埋H&E染色后观察脓毒症肾损伤的程度，并检测血清肌酐含量。结果:不同浓度脂多糖（LPS）刺激血小板后，血小板分泌的细胞因子IL-1β、IL-6增加；血小板和肾小管上皮细胞共培养体系中，p-p65蛋白含量增加，TAK-242抑制血小板活化，减少肾小管上皮细胞炎症因子分泌；构建小鼠盲肠结扎穿孔的脓毒症模型后，尾静脉注射TAK-242，抑制血小板表面TLR4的激活，肾小管增殖能力和病理改善，血清肌酐降低。结论:脓毒症患者中血小板聚集并激活，分泌炎症因子增加，激活肾小管上皮细胞NF-κB信号通路，导致肾损伤，TLR4抑制剂可改善肾损伤，有潜在临床应用价值。

关键词: 脓毒症, 肾损伤, 血小板活化, NF-κB信号通路

Abstract:

AIM: To investigate the effect of platelet activation on renal tubular epithelial cells in sepsis. METHODS: ELISA was used to detect the inflammatory cytokines IL-1β and IL-6 secreted by platelets. Renal tubular epithelial cells and platelets were co-cultured in the co-culture system. Western blot was used to test p65 and p-p65 protein in renal tubular epithelial cells in the lower chamber. IL-1β and IL-6 mRNA expression level was measured by qRT-PCR. Cecal ligation and puncture(CLP) were used to construct the sepsis model, and TLR4 inhibitor TAK-242 was injected in caudal vein. In order to observe injury degree of renal tissue, EdU experiment and serum creatinine detection were carried out. RESULTS: IL-1β and IL-6 cytokines secreted by active platelets were increased in the presence of different concentrations of LPS. In the platelet and renal tubular epithelial cells co-culture system, p-p65 protein was increased in the tubular epithelial cells. TAK-242 could inhibit platelet activation, and reduce the renal tubular epithelial cell inflammatory cytokines secretion.The proliferation and pathological status were improved, and serum creatinine was decreased. CONCLUSION: The platelets in septic patients are aggregated and activated. inflammatory cytokines are increased. These cytokines activate NF-κB signaling pathway of renal tubular epithelial cells, leading to renal injury. TLR4 inhibitors have potential clinical value to improve the renal injury in sepsis.

Key words: sepsis, kidney injury, platelet activation, NF-κB signaling pathway

中图分类号:

R965.2

方晶晶，陶静，黄钦，史超路，颜碧清，盖磊，厉旭光. 脓毒症中血小板活化对肾小管上皮影响的机制研究[J]. 中国临床药理学与治疗学, 2018, 23(3): 283-289.

FANG Jingjing, TAO Jing, HUANG Qin, SHI Chaolu, YAN Biqing, GAI Lei, LI Xuguang. Effects of platelet activation on renal tubular epithelial cells in sepsis[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2018, 23(3): 283-289.

参考文献

［1］Fleischmann C, Scherag A, Adhikari NK, et al. Assessment of global incidence and mortality of hospital-treated sepsis: current estimates and limitations［J］. Am J Respir Crit Care Med, 2016, 193(3): 259-272.
［2］Singer M, Deutschman CS, Seymour CW, et al. The third international consensus definitions for sepsis and septic shock (sepsis-3)［J］. JAMA, 2016, 315(8): 801-810.
［3］Bagshaw SM, George C, Bellomo R, et al. Early acute kidney injury and sepsis: a multicentre evaluation［J］. Crit Care, 2008, 12(2): R47.
［4］Hoste EA, Clermont G, Kersten A, et al. RIFLE criteria for acute kidney injury are associated with hospital mortality in critically ill patients: a cohort analysis［J］. Crit Care, 2006, 10(3): R73.
［5］Tokes-Fuzesi M, Woth G, Ernyey B, et al. Microparticles and acute renal dysfunction in septic patients［J］. J Crit Care, 2013, 28(2): 141-147.
［6］Zhao WY, Zhang L, Sui MX, et al. Protective effects of sirtuin 3 in a murine model of sepsis-induced acute kidney injury［J］. Sci Rep, 2016, 6: 33201.
［7］Kraemer BF, Campbell RA, Schwertz H, et al. Bacteria differentially induce degradation of Bcl-xL, a survival protein, by human platelets［J］. Blood, 2012, 120(25): 5014-5020.
［8］Garraud O, Cognasse F. Platelet toll-like receptor expression: platelet toll-like receptor expression: the link between "danger" ligands and inflammation［J］. Inflamm Allergy Drug Targets, 2010, 9(5): 322-333.
［9］Clark SR, Ma AC, Tavener SA, et al. Platelet TLR4 activates neutrophil extracellular traps to ensnare bacteria in septic blood［J］. Nat Med, 2007, 13(4): 463-469.
［10］Rondina MT, Weyrich AS, Zimmerman GA. Platelets as cellular effectors of inflammation in vascular diseases［J］. Circ Res, 2013, 112(11): 1506-1519.
［11］Semple JW, Italiano JE, Jr., Freedman J. Platelets and the immune continuum［J］. Nat Rev Immunol, 2011, 11(4): 264-274.
［12］Andonegui G, Kerfoot SM, McNagny K, et al. Platelets express functional Toll-like receptor-4［J］. Blood, 2005, 106(7): 2417-2423.
［13］Denis MM, Tolley ND, Bunting M, et al. Escaping the nuclear confines: signal-dependent pre-mRNA splicing in anucleate platelets［J］. Cell, 2005, 122(3): 379-391.
［14］Boilard E, Nigrovic PA, Larabee K, et al. Platelets amplify inflammation in arthritis via collagen-dependent microparticle production［J］. Science, 2010, 327(5965): 580-583.
［15］Cloutier N, Pare A, Farndale RW, et al. Platelets can enhance vascular permeability［J］. Blood, 2012, 120(6): 1334-1343.
［16］El-Achkar TM, Hosein M, Dagher PC. Pathways of renal injury in systemic gram-negative sepsis［J］. Eur J Clin Invest, 2008, 38 Suppl 2: 39-44.
［17］Fenhammar J, Rundgren M, Hultenby K, et al. Renal effects of treatment with a TLR4 inhibitor in conscious septic sheep［J］. Crit Care, 2014, 18(5): 488.
［18］朱晓玲, 王永钧, 杨汝春,等. 复方积雪草有效组分干预肾小管上皮细胞Toll样受体4表达的实验研究［J］. 中国临床药理学与治疗学, 2009, 14(2):171-174.
［19］魏军, 王艳红, 蒋金辉,等. 小剂量肝素对脓毒症患者凝血功能及血小板活化水平影响［J］. 现代仪器与医疗, 2015, 21(6):10-12.

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脓毒症中血小板活化对肾小管上皮影响的机制研究

Effects of platelet activation on renal tubular epithelial cells in sepsis

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