摘要
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、24h,3 组间血清肌酐(F=405.699、485.730、1 696.551、2055.637,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.
Abstract
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=405.699,485.730,1 696.551,2055.637;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.
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