摘要
目的探讨山莨菪碱是否可以调节复苏后动物辅助性T细胞/调节性T细胞比值(Th17/Treg比值)及其对机体的保护作用。方法将24只北京白小型猪随机分为假手术组(Sham组)、心室颤动(室颤)除颤+生理盐水组(SA组)、室颤除颤+氢溴酸山莨菪碱组(AH组),每组8只。SA组和AH组通过心室内电极持续刺激诱发室颤8 min并复苏制备缺血/再灌注(I/R)模型,其中SA组心肺复苏(CPR)后仅给予生理盐水静脉滴注(静滴),AH组CPR后给予生理盐水+氢溴酸山莨菪碱静滴。分别于心搏骤停(CA)前及自主循环恢复(ROSC)后0、1、2、4、6 h记录各组猪的血流动力学指标和动脉血气分析指标、静脉血白细胞介素(IL-17、IL-10)水平及IL-17/IL-10比值。ROSC后6 h处死动物,留取肠道淋巴组织,光镜下观察病理学改变,同时检测组织匀浆中IL-17和IL-10水平(以IL-17/IL-10比值代表Th17/Treg比值)评估复苏猪的免疫状态,并对组织进行培养以评估细菌移位情况。结果随ROSC时间延长,SA组和AH组静脉血IL-17及IL-17/IL-10比值均呈明显降低趋势,IL-10水平呈明显升高趋势;且从ROSC后2 h开始,AH组IL-17/IL-10比值均明显高于SA组,持续至ROSC后6 h(0.79±0.05比0.49±0.08,P<0.05)。光镜下显示,与SA组相比,AH组肠淋巴组织皮质区淋巴小结数量较少、体积较小。ROSC后6 h,与Sham组比较,SA组和AH组猪肠道淋巴组织内IL-17及IL-17/IL-10比值均明显降低〔IL-17(ng/L):155.23±0.92、178.76±7.25比209.21±19.82,IL-17/IL-10比值:1.43±0.13、1.92±0.18比3.30±0.31,均P<0.05〕,IL-10均明显升高(ng/L:109.85±11.60、93.55±81.83比63.45±0.62,均P<0.05);而AH组IL-17/IL-10比值明显高于SA组(1.92±0.18比1.43±0.13,P<0.05)。细菌培养结果显示,SA组和AH组猪复苏后均出现肠道细菌移位,提示复苏后动物出现免疫抑制,肠源性继发感染风险升高;而AH组肠道淋巴组织细菌培养率明显低于SA组〔62.5%(5/8)比87.5%(7/8),P<0.05〕,提示山莨菪碱可降低细菌移位风险。结论山莨菪碱可以通过影响复苏动物Th17/Treg细胞因子平衡来发挥免疫调节作用,以减少肠源性继发感染的风险,具有器官保护作用。
Objective To investigate whether anisodamine can regulate the ratio of helper T helper cells/regulatory T cells(Th17/Treg)and its protective effect on animals after resuscitation.Methods Twenty-four Beijing white minipigs were randomly divided into sham operation group(Sham group),resuscitation and normal saline group(SA group),and resuscitation and anisodamine hydrobromide group(AH group),with 8 pigs in each group.In SA group and AH group,ventricular fibrillation was induced by continuous stimulation with intraventricular electrodes for 8 minutes and then resuscitated to establish ischemia/reperfusion(I/R)model.In SA group,after cardiopulmonary resuscitation(CPR),only normal saline was intravenously infused,while in AH group,normal saline and anisodamine hydrobromide were given intravenously at the same time point.Hemodynamic indexes,arterial blood gas analysis indexes,interleukins(IL-17,IL-10)levels in venous blood and IL-17/IL-10 ratio were recorded at 6 different time points:baseline,immediately after return of spontaneous circulation(ROSC),1 hour,2 hours,4 hours and 6 hours after ROSC.The animals were sacrificed at 6 hours after ROSC,and intestinal lymphatic tissues were taken to observe pathological changes under light microscope.At the same time,the levels of IL-17 and IL-10 in intestinal lymphatic tissue were measured(the ratio of IL-17/IL-10 represents the ratio of Th17/Treg cytokines)to evaluate the immune status of the resuscitated animals.The bacterial translocations of different groups were evaluated by culturing intestinal lymphoid tissue.Results With the extension of ROSC time,the levels of IL-17 in venous blood and the IL-17/IL-10 ratio in pig blood samples continued to decrease,while the levels of IL-10 continued to increase.From 2 hours after ROSC,the IL-17/IL-10 ratio in AH group was significantly higher than that in SA group continued until at 6 hours after ROSC(0.79±0.05 vs.0.49±0.08,P<0.05).Light microscopy showed that the number and size of lymph nodules in the cortex of intestinal lymphatic tissue were less in AH group,compared with SA group.Compared with Sham group,the levels of IL-17 and IL-17/IL-10 ratio also decreased in intestinal lymphatic tissue at 6 hours after ROSC[IL-17(ng/L):155.23±0.92,178.76±7.25 vs.209.21±19.82,IL-17/IL-10 ratio:1.43±0.13,1.92±0.18 vs.3.30±0.31,all P<0.05],and IL-10 increased significantly(ng/L:109.85±11.60,93.55±81.83 vs.63.45±0.62,all P<0.05);IL-17/IL-10 ratio in AH group was significantly higher than that in SA group(1.92±0.18 vs.1.43±0.13,P<0.05).Tissue culture indicated the intestinal bacterial translocation after resuscitation,suggesting that the animals had immunosuppression and the increased risk of intestinal secondary infection after resuscitation.Compared with SA group,the risk of bacterial translocation was lower than that in AH group[62.5%(5/8)vs.87.5%(7/8),P<0.05].Conclusions Anisodamine plays an immunomodulatory role by affecting the balance of Th17/Treg cytokines in resuscitated animals,so as to reduce the risk of intestinal secondary infection and has an organ protective effect.
作者
董桂娟
杨军
赵鑫
梅雪
Dong Guijuan;Yang Jun;Zhao Xin;Mei Xue(Emergency Medicine Clinical Research Center,Beijing Chao-Yang Hospital,Capital Medicine University,Beijing Key Laboratory of Cardiopulmonary Cerebral Resuscitation,Beijing 100020,China)
出处
《中华危重病急救医学》
CAS
CSCD
北大核心
2022年第9期964-969,共6页
Chinese Critical Care Medicine
基金
睿E(睿意)急诊医学研究专项基金(R2019013)。
关键词
心肺复苏
TH细胞
氢溴酸山莨菪碱
细菌移位
Cardiopulmonary resuscitation
Th cell
Anisodamine hydrobromide
Bacterial translocation
