切换至 "中华医学电子期刊资源库"

中华危重症医学杂志(电子版) ›› 2023, Vol. 16 ›› Issue (06) : 441 -447. doi: 10.3877/cma.j.issn.1674-6880.2023.06.001

论著

西维来司钠对猪心肺复苏后肾肠损伤作用的研究
周腾达, 陈庆丽, 杨雪林, 陈琪, 徐杰丰, 周光居, 张茂()   
  1. 310009 杭州,浙江大学医学院附属第二医院急诊医学科/浙江省严重创伤与烧伤诊治重点实验室/浙江省急危重症临床医学研究中心;现工作单位为宁波市第二医院急诊医学科
    323000 浙江丽水,丽水市人民医院急诊医学科
    323000 浙江丽水,丽水市中心医院急诊医学科
    311400 杭州,杭州市富阳区第一人民医院重症医学科
    310009 杭州,浙江大学医学院附属第二医院急诊医学科/浙江省严重创伤与烧伤诊治重点实验室/浙江省急危重症临床医学研究中心
  • 收稿日期:2023-06-16 出版日期:2023-12-31
  • 通信作者: 张茂
  • 基金资助:
    国家自然科学基金项目(82072126); 浙江省重点研发计划项目(2021C03073、2021C03036)

Effect of sivelestat on renal and intestinal injuries after cardiopulmonary resuscitation in pigs

Tengda Zhou, Qingli Chen, Xuelin Yang, Qi Chen, Jiefeng Xu, Guangju Zhou, Mao Zhang()   

  1. Department of Emergency Medicine, the Second Affiliated Hospital, Zhejiang University School of Medicine / Zhejiang Provincial Key Laboratory for Diagnosis and Treatment of Severe Trauma and Burn/Zhejiang Provincial Acute and Critical Care Clinical Medical Research Center, Hangzhou 310009, China
    Department of Emergency Medicine, Lishui People's Hospital, Lishui 323000, China
    Department of Emergency Medicine, Lishui Central Hospital, Lishui 323000, China
    Department of Intensive Care, the First People's Hospital of Fuyang District, Hangzhou 311400, China
  • Received:2023-06-16 Published:2023-12-31
  • Corresponding author: Mao Zhang
引用本文:

周腾达, 陈庆丽, 杨雪林, 陈琪, 徐杰丰, 周光居, 张茂. 西维来司钠对猪心肺复苏后肾肠损伤作用的研究[J/OL]. 中华危重症医学杂志(电子版), 2023, 16(06): 441-447.

Tengda Zhou, Qingli Chen, Xuelin Yang, Qi Chen, Jiefeng Xu, Guangju Zhou, Mao Zhang. Effect of sivelestat on renal and intestinal injuries after cardiopulmonary resuscitation in pigs[J/OL]. Chinese Journal of Critical Care Medicine(Electronic Edition), 2023, 16(06): 441-447.

目的

通过建立猪心肺复苏(CPR)模型,探讨在猪复苏成功后使用西维来司钠对肾肠损伤的保护效果及机制。

方法

将25头健康白猪分成假手术组(n = 6)、CPR组(n = 10)和西维来司钠组(n = 9)。假手术组只进行实验造模准备,另两组经右心室放电诱导心室颤动持续9 min后开始CPR 6 min以建立动物模型。复苏成功后5 min,西维来司钠组经股静脉泵入西维来司钠10 mg/kg持续1 h,其余两组泵入等量等渗NaCl溶液。在诱颤前及复苏后1、2、4、24 h依次采血检测血清肌酐、尿素氮(BUN)、肠型脂肪酸结合蛋白(iFABP)以及二胺氧化酶(DAO)含量。在复苏后24 h将动物安乐死后立即取肾肠组织标本,测量组织中NOD样受体蛋白3(NLRP3)、裂解的半胱氨酸天冬氨酸蛋白酶1(C-CAS-1)、消皮素D(GSDMD)蛋白表达情况,以及组织中白细胞介素18(IL-18)和IL-1β炎症因子水平。

结果

诱颤前,3组动物间血清肌酐、BUN、iFABP、DAO比较,差异均无统计学意义(F = 0.474、0.722、0.249、1.465,P = 0.629、0.497、0.781、0.253)。复苏后1、2、4、24 h,3组间血清肌酐(F = 405.699、485.730、1 696.551、2 055.637,P均< 0.001)、BUN(F = 98.617、354.974、451.838、1 139.201,P均< 0.001)、iFABP含量(F = 49.349、115.131、283.316、510.195,P均< 0.001)及DAO活性(F = 35.020、105.490、378.235、543.949,P均< 0.001)比较,差异均有统计学意义。CPR组及西维来司钠组血清肌酐、BUN及DAO活性复苏后1、2、4、24 h较假手术组均明显升高,且CPR组更高(P均< 0.05)。而西维来司钠组仅在复苏后2、4、24 h的血清iFABP含量较CPR组均明显降低(P均< 0.05)。同时,复苏后3组动物肾肠组织NLRP3(F = 13.937、47.761,P均< 0.001)、C-CAS-1(F = 64.209、48.423,P均< 0.001)、GSDMD(F = 14.583、93.754,P均< 0.001)蛋白表达水平及IL-1β(F = 566.930、171.428,P均< 0.001)、IL-18(F = 619.730、84.092,P均< 0.001)含量的比较,差异均有统计学意义。且进一步两两比较发现,肾肠组织中西维来司钠组动物的NLRP3、C-CAS-1、GSDMD蛋白表达水平及IL-1β与IL-18含量均显著低于CPR组(P均< 0.05)。

结论

在猪CPR后早期给予西维来司钠对肾肠组织损伤有保护作用,可能与减少肾肠组织细胞焦亡有关。

Objective

To investigate the protective effect and mechanism of sivelestat on renal and intestinal injury after cardiopulmonary resuscitation (CPR) in pigs.

Methods

A total of 25 healthy white pigs were randomly divided into a sham group (n = 6), a CPR group (n = 10) and a sivelestat group (n = 9). The pigs in the sham group were only used for experimental preparation; the pigs in other two groups were given ventricular fibrillation induced by right ventricular discharge for 9 min and then CPR for 6 min. At 5 min after successful resuscitation, the pigs in the sivelestat group were given 10 mg/kg sivelestat through the femoral vein for 1 h, and the other two groups were given the same amount of isotonic NaCl solution. Blood samples were collected before ventricular fibrillation and at 1, 2, 4 and 24 h after resuscitation to detect serum creatinine, blood urea nitrogen (BUN), intestinal fatty acid-binding protein (iFABP) and diamine oxidase (DAO). At 24 h after resuscitation, the pigs were euthanized and the tissue samples of kidney and intestine were taken immediately to measure the protein expression levels of NOD like receptor protein 3 (NLRP3), cleaved caspase-1 (C-CAS-1) and Gasdermin D (GSDMD), as well as the levels of interleukin-18 (IL-18) and IL-1β.

Results

There were no significant differences in the serum creatinine, BUN, iFABP and DAO among the sham group, CPR group and sivelestat group before ventricular fibrillation (F = 0.474, 0.722, 0.249, 1.465; P = 0.629, 0.497, 0.781, 0.253). The serum levels of creatinine (F = 405.699, 485.730, 1 696.551, 2 055.637; all P < 0.001), BUN (F = 98.617, 354.974, 451.838, 1 139.201; all P < 0.001), iFABP (F = 49.349, 115.131, 283.316, 510.195; all P < 0.001) and DAO (F = 35.020, 105.490, 378.235, 543.949; all P < 0.001) were significantly different among the three groups at 1, 2, 4 and 24 h after resuscitation. The serum levels of creatinine, BUN and DAO in the CPR group and sivelestat group were significantly higher than those in the sham group at 1, 2, 4 and 24 h after resuscitation, and they were highest in the CPR group (all P < 0.05). However, the serum iFABP levels in the sivelestat group were significantly lower than those in the CPR group only at 2, 4 and 24 h after resuscitation (all P < 0.05). Meanwhile, the protein expression levels of NLRP3 (F = 13.937, 47.761; both P < 0.001), C-CAS-1 (F = 64.209, 48.423; both P < 0.001), GSDMD (F = 14.583, 93.754; both P < 0.001) and the contents of IL-1β (F = 566.930, 171.428; both P < 0.001), IL-18 (F = 619.730, 84.092; both P < 0.001) in renal and intestinal tissues among the three groups all showed significant differences after resuscitation. Further pairwise comparison found that the protein expression levels of NLRP3, C-CAS-1, GSDMD and the contents of IL-1β, IL-18 in the sivelestat group were significantly lower than those in the CPR group (all P < 0.05).

Conclusion

Early application of sivelestat after CPR in pigs can inhibit pyroptosis reaction and reduce the damage of kidney and intestine.

表1 各组动物诱颤前及复苏后各时间点血清肌酐和BUN水平的比较( ± s
表2 各组动物诱颤前及复苏后各时间点血清iFABP含量和DAO活性的比较( ± s
图1 Western-blotting法检测各组动物肾肠组织NLRP3、C-CAS-1、GSDMD蛋白表达情况注:NLRP3. NOD样受体蛋白3;C-CAS-1.裂解的半胱氨酸天冬氨酸蛋白酶1;GSDMD.消皮素D;GAPDH. 3-磷酸甘油醛脱氢酶;CPR.心肺复苏
表3 各组动物复苏后肾肠组织NLRP3、C-CAS-1、GSDMD蛋白表达水平比较( ± s
表4 各组动物复苏后肾肠组织IL-1β与IL-18含量的比较(ng/L, ± s
1
朱威,徐佳,陆远强.《2020年美国心脏协会心肺复苏及心血管急救指南》成人生命支持部分建议内容分析[J/CD].中华危重症医学杂志(电子版)2020,13(5):379-381.
2
Chelly J, Mpela AG, Jochmans S, et al. OHCA (out-of-hospital cardiac arrest) and CAHP (cardiac arrest hospital prognosis) scores to predict outcome after in-hospital cardiac arrest: insight from a multicentric registry[J]. Resuscitation, 2020 (156): 167-173.
3
Patyna S, Riekert K, Buettner S, et al. Acute kidney injury after in-hospital cardiac arrest in a predominant internal medicine and cardiology patient population: incidence, risk factors, and impact on survival[J]. Ren Fail, 2021, 43 (1): 1163-1169.
4
Dutta A, Hari KJ, Azizian J, et al. Incidence, predictors, and prognosis of acute kidney injury among cardiac arrest survivors[J]. J Intensive Care Med, 2021, 36 (5): 550-556.
5
Chalkias A, Scheetz MH, Gulati A, et al. Periarrest intestinal bacterial translocation and resuscitation outcome[J]. J Crit Care, 2016, 31 (1): 217-220.
6
Matsuzaki K, Hiramatsu Y, Homma S, et al. Sivelestat reduces inflammatory mediators and preserves neutrophil deformability during simulated extracorporeal circulation[J]. Ann Thorac Surg, 2005, 80 (2): 611-617.
7
Ishikawa N, Oda M, Kawaguchi M, et al. The effects of a specific neutrophil elastase inhibitor (ONO-5046) in pulmonary ischemia-reperfusion injury[J]. Transpl Int, 2003, 16 (5): 341-346.
8
Takayama M, Ishibashi M, Ishii H, et al. Effects of neutrophil elastase inhibitor (ONO-5046) on lung injury after intestinal ischemia-reperfusion[J]. J Appl Physiol (1985), 2001, 91 (4): 1800-1807.
9
Nolan JP, Neumar RW, Adrie C, et al. Post-cardiac arrest syndrome: epidemiology, pathophysiology, treatment, and prognostication. A Scientific Statement from the International Liaison Committee on Resuscitation; the American Heart Association Emergency Cardiovascular Care Committee; the Council on Cardiovascular Surgery and Anesthesia; the Council on Cardiopulmonary, Perioperative, and Critical Care; the Council on Clinical Cardiology; the Council on Stroke[J]. Resuscitation, 2008, 79 (3): 350-379.
10
Schofield ZV, Woodruff TM, Halai R, et al. Neutrophils—a key component of ischemia-reperfusion injury[J]. Shock, 2013, 40 (6): 463-470.
11
El-Benna J, Hurtado-Nedelec M, Marzaioli V, et al. Priming of the neutrophil respiratory burst: role in host defense and inflammation[J]. Immunol Rev, 2016, 273 (1): 180-193.
12
Fialkow L, Wang Y, Downey GP. Reactive oxygen and nitrogen species as signaling molecules regulating neutrophil function[J]. Free Radic Biol Med, 2007, 42 (2): 153-164.
13
Kaplan MJ, Radic M. Neutrophil extracellular traps: double-edged swords of innate immunity[J]. J Immunol, 2012, 189 (6): 2689-2695.
14
录亚鹏,刘洋,郝伟,等.细胞焦亡在肠缺血再灌注损伤中作用的研究进展[J].中国临床药理学与治疗学2021,26(12):1437-1443.
15
吴燕升,高建东.细胞焦亡在肾脏炎性损伤中的作用研究进展[J].基础医学与临床2017,37(10):1470-1473.
16
Rathinam VAK, Zhao Y, Shao F. Innate immunity to intracellular LPS[J]. Nat Immunol, 2019, 20 (5): 527-533.
17
Brandon EB, Ashley NE, Hua Z, et al. Gasdermin D in pyroptosis[J]. Acta Pharmaceutica Sinica B, 2021, 11 (9): 2768-2782.
18
Iyer SS, Pulskens WP, Sadler JJ, et al. Necrotic cells trigger a sterile inflammatory response through the Nlrp3 inflammasome[J]. Proc Natl Acad Sci U S A, 2009, 106 (48): 20388-20393.
19
Wang Z, Li Z, Feng D, et al. Autophagy induction ameliorates inflammatory responses in intestinal ischemia-reperfusion through inhibiting NLRP3 inflammasome activation[J]. Shock, 2019, 52 (3): 387-395.
20
Kawabata K, Hagio T, Matsuoka S. Pharmacological profile of a specific neutrophil elastase inhibitor, Sivelestat sodium hydrate[J]. Nihon Yakurigaku Zasshi, 2003, 122 (2): 151-160.
21
Hayama T, Matsuyama M, Funao K, et al. Benefical effect of neutrophil elastase inhibitor on renal warm ischemia-reperfusion injury in the rat[J]. Transplant Proc, 2006, 38 (7): 2201-2202.
22
Nakamura N, Morisaki H, Suzuki T, et al. Inhibition of neutrophil elastase attenuates gut mucosal injury evoked by acute alveolar hypoxia in rabbits[J]. Shock, 2007, 28 (1): 101-105.
23
Bonaventura A, Vecchié A, Abbate A, et al. Neutrophil extracellular traps and cardiovascular diseases: an update[J]. Cells, 2020, 9 (1): 231.
24
Hu Q, Shi H, Zeng T, et al. Increased neutrophil extracellular traps activate NLRP3 and inflammatory macrophages in adult-onset Still's disease[J]. Arthritis Res Ther, 2019, 21 (1): 9.
25
Okeke EB, Louttit C, Fry C, et al. Inhibition of neutrophil elastase prevents neutrophil extracellular trap formation and rescues mice from endotoxic shock[J]. Biomaterials, 2020 (238): 119836.
[1] 许彬, 王丽, 陈瑞, 沈奕, 陆件. 瞬时受体电位粘脂素1介导细胞自噬在远端缺血后处理保护大鼠脑缺血-再灌注损伤中的作用研究[J/OL]. 中华危重症医学杂志(电子版), 2024, 17(03): 180-187.
[2] 黄蓉, 梁自毓, 祁文瑾. NLRP3炎症小体在胎膜早破孕妇血清中的表达及其意义[J/OL]. 中华妇幼临床医学杂志(电子版), 2024, 20(05): 540-548.
[3] 李嘉兴, 孙乙文, 李文星. NLRP3炎性小体在急性胰腺炎中作用的研究进展[J/OL]. 中华普通外科学文献(电子版), 2024, 18(04): 300-304.
[4] 唐亦骁, 陈峻, 连正星, 胡海涛, 鲁迪, 徐骁, 卫强. 白果内酯对小鼠肝缺血再灌注损伤保护作用研究[J/OL]. 中华移植杂志(电子版), 2024, 18(05): 278-282.
[5] 王雪玲, 曹欢, 顾劲扬. 肠道菌群在器官缺血再灌注损伤中的作用及机制研究进展[J/OL]. 中华移植杂志(电子版), 2024, 18(04): 247-250.
[6] 彭瑞, 杨瑞文, 魏澹宁, 夏永良. 琥珀酸受体1加重肾脏缺血再灌注损伤的作用研究[J/OL]. 中华移植杂志(电子版), 2024, 18(03): 159-164.
[7] 孙璐, 蒋亚玲, 陈凌君. 布托啡诺对脑缺血再灌注损伤大鼠神经炎症和JAK2/STAT3信号通路的影响[J/OL]. 中华细胞与干细胞杂志(电子版), 2024, 14(06): 344-350.
[8] 阿卜杜萨拉木·图尔荪麦麦提, 吐尔洪江·吐逊, 温浩. 肝脏缺血-再灌注损伤与cGAS-STING信号通路[J/OL]. 中华肝脏外科手术学电子杂志, 2024, 13(03): 394-397.
[9] 张云飞, 吐尔洪江·吐逊. NLRP3炎症小体及其在肝脏缺血-再灌注损伤中的作用机制[J/OL]. 中华肝脏外科手术学电子杂志, 2024, 13(03): 398-403.
[10] 张一绚, 韩冰, 刘超, 李思晨, 孙雪峰. 年轻化内环境改善老年小鼠肾缺血再灌注损伤诱导的肾间质纤维化[J/OL]. 中华肾病研究电子杂志, 2024, 13(03): 129-133.
[11] 李京, 牛博, 刘晓蓓, 魏新雪, 黄荣. circ-SESN2 沉默靶向调控miRNA-23a-5p/ULK1 在神经细胞氧化应激损伤中的作用机制研究[J/OL]. 中华神经创伤外科电子杂志, 2024, 10(05): 263-272.
[12] 丁洪基, 赵长江, 孙鹏飞, 王灿, 王贵珍, 李龙龙. 细胞焦亡与疾病的关系研究进展[J/OL]. 中华临床医师杂志(电子版), 2024, 18(07): 682-686.
[13] 刘俊彬, 张晓婷, 郭镜培, 刘佳妮, 于本帅, 张可, 周斌. 熊果酸通过抑制NLRP3介导的小胶质细胞焦亡减轻脑缺血再灌注损伤的研究[J/OL]. 中华介入放射学电子杂志, 2024, 12(03): 221-227.
[14] 郭子宾, 柯学锋, 余琳潇, 张伟艺, 张军, 汪娟. 2015年至2019年盐田区院外心脏骤停患者救治成功的影响因素分析和应对策略[J/OL]. 中华卫生应急电子杂志, 2024, 10(04): 199-202.
[15] 刘晴雯, 韩勇, 陈丽丹, 邓哲. 早期机械通气对成人院内心脏骤停病死率的影响:一项回顾性队列研究[J/OL]. 中华卫生应急电子杂志, 2024, 10(04): 203-206.
阅读次数
全文


摘要