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Article(id=1149768567169659864, tenantId=1146029695717560320, journalId=1146123166801305609, issueId=1149768563956826506, articleNumber=null, orderNo=null, doi=10.12404/j.issn.1671-1815.2406882, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1726156800000, receivedDateStr=2024-09-13, revisedDate=1742745600000, revisedDateStr=2025-03-24, acceptedDate=null, acceptedDateStr=null, onlineDate=1752055788080, onlineDateStr=2025-07-09, pubDate=1749312000000, pubDateStr=2025-06-08, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1752055788080, onlineIssueDateStr=2025-07-09, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1752055788080, creator=13701087609, updateTime=1752055788080, updator=13701087609, issue=Issue{id=1149768563956826506, tenantId=1146029695717560320, journalId=1146123166801305609, year='2025', volume='25', issue='16', pageStart='6587', pageEnd='7021', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1752055787314, creator=13701087609, updateTime=1768456850262, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1218559607937618069, tenantId=1146029695717560320, journalId=1146123166801305609, issueId=1149768563956826506, language=EN, specialIssueTitle=, coverIllustrator=, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1218559607937618070, tenantId=1146029695717560320, journalId=1146123166801305609, issueId=1149768563956826506, language=CN, specialIssueTitle=, coverIllustrator=, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=6674, endPage=6681, ext={EN=ArticleExt(id=1149768567408735193, articleId=1149768567169659864, tenantId=1146029695717560320, journalId=1146123166801305609, language=EN, title=Modification and Evaluation of an Improved Asphyxia Method for Constructing a Rat Model of Global Cerebral Ischemia-reperfusion Injury Following Cardiac Arrest and Cardiopulmonary Resuscitation, columnId=1156262732384031076, journalTitle=Science Technology and Engineering, columnName=Papers·Medicine, runingTitle=null, highlight=null, articleAbstract=

Cardiac arrest and global cerebral ischemia-reperfusion injury after cardiopulmonary resuscitation (CPR) are common pathological conditions in critically ill patients, with high mortality and disability rates. Currently, the conventional asphyxia method is commonly used to establish a brain injury model after CPR. However, it has a low resuscitation success rate and postoperative survival rate. Therefore, a modified asphyxia model of cardiac arrest and global cerebral ischemia-reperfusion injury after CPR was established and compared it with the conventional asphyxia method to evaluate its advantages. Sprague Dawley(SD) rats were randomly divided into a conventional group and a modified group. The conventional asphyxia method and the modified asphyxia method were used to establish the models, respectively. The resuscitation success rate and 24-hour survival rate were observed. Neurological deficits were assessed using neurological deficit scores. Hematoxylin-eosin staining was used to observe pathological changes in brain tissue. Transmission electron microscopy was used to examine neuronal ultrastructure. Western Blot analysis was performed to detect inflammatory, oxidative stress, and apoptotic markers. The results show that compared with the conventional asphyxia method, the modified asphyxia method has a higher resuscitation success rate and 24-hour survival rate. No significant differences are observed in brain injury, inflammation, oxidative stress, and apoptosis between the two methods. The modified asphyxia method meets the requirements for studying global cerebral ischemia-reperfusion injury.

, correspAuthors=Ying LIU, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, authorCompany=null, fund=null, authors=null, authorsList=Ou-yang RAO, Shi-xin LI, Jie HU, Sha XIE, Ying LIU), CN=ArticleExt(id=1149768576447459403, articleId=1149768567169659864, tenantId=1146029695717560320, journalId=1146123166801305609, language=CN, title=窒息法构建大鼠心脏骤停-心肺复苏后全脑缺血再灌注损伤模型的改良与评价, columnId=1156262732526637414, journalTitle=科学技术与工程, columnName=论文·医药、卫生, runingTitle=null, highlight=null, articleAbstract=

心脏骤停-心肺复苏后全脑缺血再灌注损伤是急危重症常见的病理损伤,其致死率和致残率较高。目前,传统窒息法常用于构建心脏骤停-心肺复苏后脑损伤模型,但其复苏成功率和术后存活率较低。为此,通过改良传统窒息法构建心脏骤停-心肺复苏后全脑缺血再灌注损伤模型,并与传统窒息法进行比较,评估改良窒息法的优势。将SD(Sprague Dawley)大鼠随机分为传统组与改良组,分别采用传统窒息法和改良窒息法构建模型。观察大鼠复苏成功率和24 h生存率,采用神经功能损害评分评估神经功能,苏木精-伊红染色观察脑组织病理改变,透射电镜观察神经元超微结构,Western Blot检测炎症、氧化应激、凋亡指标。结果表明:与传统窒息法相比,改良窒息法构建大鼠心脏骤停-心肺复苏后脑缺血再灌注损伤模型具有更高的复苏成功率和24 h存活率,且脑损伤、炎症、氧化应激、凋亡等情况与传统窒息法比较无明显差异,能够满足全脑缺血再灌注损伤研究的需要。

, correspAuthors=刘颖, authorNote=null, correspAuthorsNote=
* 刘颖(1976—),女,汉族,贵州贵阳人,博士,主任医师。研究方向:脑缺血再灌注损伤机制。E-mail:。
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饶欧阳(1997—),男,汉族,四川德阳人,硕士研究生。研究方向:脑缺血再灌注损伤机制。E-mail:。

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饶欧阳(1997—),男,汉族,四川德阳人,硕士研究生。研究方向:脑缺血再灌注损伤机制。E-mail:。

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饶欧阳(1997—),男,汉族,四川德阳人,硕士研究生。研究方向:脑缺血再灌注损伤机制。E-mail:。

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*表示P<0.05

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*表示P<0.05;**表示P<0.01,差异具有统计学意义

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Modeling information of the traditional group and the modified group

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分组 传统组 改良组
大鼠数量/只 38 52
大鼠体重/g 250~350 250~300
气管插管 间接照明经口直
视下插管		 可视喉镜经口直视下
插管+气管导管固定
动静脉插管 PE50导管 PE50导管
心电监护 — Ⅱ导联心电监护
胸外按压/
(次·min-1)		 手动胸外按压250 节拍器引导下手动
胸外按压200
肾上腺素 0.01 mg、2 min
重复一次		 0.02 mg、2 min
重复一次
机械通气 频率70次/min,潮
气量0.7 mL/100 g		 频率70次/min,潮气
量2 mL/100 g
术后营养支持 — 皮下注射5%葡萄糖
注射液1 mL
), ArticleFig(id=1177980932255531866, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1149768567169659864, language=CN, label=表1, caption=

传统组与改良组造模信息

, figureFileSmall=null, figureFileBig=null, tableContent=
分组 传统组 改良组
大鼠数量/只 38 52
大鼠体重/g 250~350 250~300
气管插管 间接照明经口直
视下插管		 可视喉镜经口直视下
插管+气管导管固定
动静脉插管 PE50导管 PE50导管
心电监护 — Ⅱ导联心电监护
胸外按压/
(次·min-1)		 手动胸外按压250 节拍器引导下手动
胸外按压200
肾上腺素 0.01 mg、2 min
重复一次		 0.02 mg、2 min
重复一次
机械通气 频率70次/min,潮
气量0.7 mL/100 g		 频率70次/min,潮气
量2 mL/100 g
术后营养支持 — 皮下注射5%葡萄糖
注射液1 mL
), ArticleFig(id=1177980932318446427, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1149768567169659864, language=EN, label=Table 2, caption=

Neurological function deficit scale

, figureFileSmall=null, figureFileBig=null, tableContent=
评分项目 评估
情况		 各项分
值/分		 总分
值/分
意识 可自主探索 — 0 20
无自主探索 — 20
呼吸 正常 — 0 20
异常 — 20
嗅觉(食物) 存在 0
消失 4
视觉(跟随手运动) 存在 0
消失 4
颅神经
反射		 听觉(对拍手反应) 存在 0 20
消失 4
角膜反射 存在 0
消失 4
胡须运动 存在 0分
消失 4分
正常 — 0
运动功能 僵硬 — 5 10
瘫痪 — 10
感觉功能
(掐尾)		 正常 — 0 10
消失 — 10
平衡木实验 存在 0
消失 5
提尾反射 存在 0
协调功能 消失 5 20
翻正反射 存在 0
消失 5
桌子边缘实验 存在 0
消失 5
), ArticleFig(id=1177980932423304028, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1149768567169659864, language=CN, label=表2, caption=

神经功能损害评分

, figureFileSmall=null, figureFileBig=null, tableContent=
评分项目 评估
情况		 各项分
值/分		 总分
值/分
意识 可自主探索 — 0 20
无自主探索 — 20
呼吸 正常 — 0 20
异常 — 20
嗅觉(食物) 存在 0
消失 4
视觉(跟随手运动) 存在 0
消失 4
颅神经
反射		 听觉(对拍手反应) 存在 0 20
消失 4
角膜反射 存在 0
消失 4
胡须运动 存在 0分
消失 4分
正常 — 0
运动功能 僵硬 — 5 10
瘫痪 — 10
感觉功能
(掐尾)		 正常 — 0 10
消失 — 10
平衡木实验 存在 0
消失 5
提尾反射 存在 0
协调功能 消失 5 20
翻正反射 存在 0
消失 5
桌子边缘实验 存在 0
消失 5
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窒息法构建大鼠心脏骤停-心肺复苏后全脑缺血再灌注损伤模型的改良与评价
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饶欧阳 1 , 李世欣 1 , 胡杰 1 , 谢沙 1 , 刘颖 1, 2, *
科学技术与工程 | 论文·医药、卫生 2025,25(16): 6674-6681
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科学技术与工程 | 论文·医药、卫生 2025, 25(16): 6674-6681
窒息法构建大鼠心脏骤停-心肺复苏后全脑缺血再灌注损伤模型的改良与评价
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饶欧阳1 , 李世欣1, 胡杰1, 谢沙1, 刘颖1, 2, *
作者信息
  • 1 贵州医科大学临床医学院, 贵阳 550004
  • 2 贵州医科大学附属医院重症医学科, 贵阳 550004

    • 饶欧阳(1997—),男,汉族,四川德阳人,硕士研究生。研究方向:脑缺血再灌注损伤机制。E-mail:。

通讯作者:

* 刘颖(1976—),女,汉族,贵州贵阳人,博士,主任医师。研究方向:脑缺血再灌注损伤机制。E-mail:。
Modification and Evaluation of an Improved Asphyxia Method for Constructing a Rat Model of Global Cerebral Ischemia-reperfusion Injury Following Cardiac Arrest and Cardiopulmonary Resuscitation
Ou-yang RAO1 , Shi-xin LI1, Jie HU1, Sha XIE1, Ying LIU1, 2, *
Affiliations
  • 1 Clinical Medical College, Guizhou Medical University, Guiyang 550004, China
  • 2 Department of Critical Care Medicine, Affiliated Hospital of Guizhou Medical University, Guiyang 550004, China
出版时间: 2025-06-08 doi: 10.12404/j.issn.1671-1815.2406882
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摘要
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心脏骤停-心肺复苏后全脑缺血再灌注损伤是急危重症常见的病理损伤,其致死率和致残率较高。目前,传统窒息法常用于构建心脏骤停-心肺复苏后脑损伤模型,但其复苏成功率和术后存活率较低。为此,通过改良传统窒息法构建心脏骤停-心肺复苏后全脑缺血再灌注损伤模型,并与传统窒息法进行比较,评估改良窒息法的优势。将SD(Sprague Dawley)大鼠随机分为传统组与改良组,分别采用传统窒息法和改良窒息法构建模型。观察大鼠复苏成功率和24 h生存率,采用神经功能损害评分评估神经功能,苏木精-伊红染色观察脑组织病理改变,透射电镜观察神经元超微结构,Western Blot检测炎症、氧化应激、凋亡指标。结果表明:与传统窒息法相比,改良窒息法构建大鼠心脏骤停-心肺复苏后脑缺血再灌注损伤模型具有更高的复苏成功率和24 h存活率,且脑损伤、炎症、氧化应激、凋亡等情况与传统窒息法比较无明显差异,能够满足全脑缺血再灌注损伤研究的需要。

全脑缺血再灌注损伤  /  心脏骤停  /  心肺复苏  /  动物模型

Cardiac arrest and global cerebral ischemia-reperfusion injury after cardiopulmonary resuscitation (CPR) are common pathological conditions in critically ill patients, with high mortality and disability rates. Currently, the conventional asphyxia method is commonly used to establish a brain injury model after CPR. However, it has a low resuscitation success rate and postoperative survival rate. Therefore, a modified asphyxia model of cardiac arrest and global cerebral ischemia-reperfusion injury after CPR was established and compared it with the conventional asphyxia method to evaluate its advantages. Sprague Dawley(SD) rats were randomly divided into a conventional group and a modified group. The conventional asphyxia method and the modified asphyxia method were used to establish the models, respectively. The resuscitation success rate and 24-hour survival rate were observed. Neurological deficits were assessed using neurological deficit scores. Hematoxylin-eosin staining was used to observe pathological changes in brain tissue. Transmission electron microscopy was used to examine neuronal ultrastructure. Western Blot analysis was performed to detect inflammatory, oxidative stress, and apoptotic markers. The results show that compared with the conventional asphyxia method, the modified asphyxia method has a higher resuscitation success rate and 24-hour survival rate. No significant differences are observed in brain injury, inflammation, oxidative stress, and apoptosis between the two methods. The modified asphyxia method meets the requirements for studying global cerebral ischemia-reperfusion injury.

global cerebral ischemia-reperfusion injury  /  cardiac arrest  /  cardiopulmonary resuscitation  /  animal model
饶欧阳, 李世欣, 胡杰, 谢沙, 刘颖. 窒息法构建大鼠心脏骤停-心肺复苏后全脑缺血再灌注损伤模型的改良与评价. 科学技术与工程, 2025 , 25 (16) : 6674 -6681 . DOI: 10.12404/j.issn.1671-1815.2406882
Ou-yang RAO, Shi-xin LI, Jie HU, Sha XIE, Ying LIU. Modification and Evaluation of an Improved Asphyxia Method for Constructing a Rat Model of Global Cerebral Ischemia-reperfusion Injury Following Cardiac Arrest and Cardiopulmonary Resuscitation[J]. Science Technology and Engineering, 2025 , 25 (16) : 6674 -6681 . DOI: 10.12404/j.issn.1671-1815.2406882
正文
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心脏骤停(cardiac arrest, CA)是一种突发的、危及生命的紧急情况,其特点是心脏突然停止有效泵血,导致全身血液循环中断[1-2],通过心肺复苏(cardiopulmonary resuscitation, CPR)成功恢复自主循环是最重要的措施。然而,心脏骤停患者心肺复苏成功后可能存在严重的脑损伤(post-cardiac arrest brain injury, PCABI),这是影响长期生存率和生活质量的关键因素[3-6]。全脑缺血再灌注损伤(global cerebral ischemia-reperfusion injury, GCIRI)是心肺复苏后脑损伤的重要机制之一,其严重性尤为显著[7-8]。GCIRI病理生理学机制复杂[9-14],目前尚不完全清楚,且临床对GCIRI治疗措施有限[15-17]。因此,GCIRI仍然是当今医学研究热点。
在医学基础研究中,建立有效的动物模型对于探索疾病机制、评估治疗方法以及开发新药具有重要意义。特别是针对心肺复苏领域,建立可靠的GCIRI动物模型有助于深入探究心脏骤停-心肺复苏后脑损伤的机制,以及探索可能的干预手段。GCIRI动物模型的构建常见的为室颤法和窒息法[18-20]。室颤法主要通过中心静脉置入绝缘导管,然后在绝缘导管中插入金属导丝,通电诱导室颤,在操作过程中还要随时调整诱颤电流大小及导丝位置,直至心电呈室颤心律,该方法对操作者技术要求及实验室设备要求较高,较难以广泛应用,而窒息法操作相对简单,对实验室设备不高,且心脏骤停-心肺复苏后脑损伤情况比室颤法更为明显,被广泛应用于GCIRI的研究,但本课题组在前期预实验阶段发现,传统的窒息法其复苏成功率及生存率较低,较难满足实验的需要。
基于上述背景,对传统窒息法[21-23]构建心脏骤停-心肺复苏后脑损伤模型进行改良,以进一步提高可操作性和复苏成功率及生存率,同时保证脑损伤效果,有助于加深对GCIRI发病机制的研究。
1 实验材料
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1.1 主要试剂与仪器
盐酸肾上腺素注射液(哈药集团三精制药有限公司)、肝素钠注射液(上海上药第一生化药业有限公司)、氯化钠注射液(四川科伦药业股份有限公司)、小动物呼吸机(ZH-3000A/B,原阳县振华教学仪器有限公司)、压力信号传感器(合肥斯派克仪器科技有限公司)、医学信号采集处理系统(ZL-620A,合肥斯派克仪器科技有限公司)。
1.2 实验动物
选用无特定病原体(specific pathogen free,SPF)级雄性SD(Sprague Dawley)大鼠,质量250~350 g,6~8周龄,由维通利华实验动物有限公司提供,许可证号:SCXK(粤)20220063。将大鼠安置在通风良好,环境安静,温度适宜(22±2)℃的SPF级动物房,进行12 h的光照/黑暗循环,自由饮食饮水,适应性喂养7 d后手术。
1.3 伦理学
动物实验方案经贵州医科大学实验动物伦理委员会批准(批准号:2304126),所有动物操作及管理均遵循中国医学科学院实验动物管理标准。
2 实验方法
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2.1 实验分组
将SD大鼠随机分为传统组与改良组,分别采用传统窒息法和改良窒息法构建心脏骤停-心肺复苏后全脑缺血再灌注损伤模型,传统组与改良组造模信息如表1所示。另选3只正常大鼠作为假手术组,仅进行气管插管,动静脉插管,不进行心脏骤停及心肺复苏操作。
2.2 造模方法
2.2.1 动物术前准备及麻醉
选用SPF级雄性SD大鼠,250~350 g,6~8周龄,用20%乌拉坦溶液(0.3 mL/100 g)腹腔注射麻醉。大鼠常规编号,备皮,仰卧位固定于手术台上,测温探头经石蜡油润滑,经肛门置入深度约3 cm后用胶布固定,造模阶段,采用控温毯保暖。
(1)气管插管。传统组的气管插管主要采用间接照明经口直视下插管或者盲插法[21-23];改良组采用自制可视喉镜下气管插管;同时,在气管插管完成后将导管用1-0缝合线妥善固定于气管管壁,如图1、图2所示。
(2)股动静脉插管。大鼠腹股沟区解剖股静脉、股动脉、股神经,如图3所示;传统组与改良组采用PE50导管插入股动静脉, PE50导管进入股动静脉约1.0 cm后在血管近心端采用4-0缝合线结收紧固定。将股动脉导管与压力传感器和医学信号采集处理系统连接,监测并记录血压波动,股静脉导管用于静脉给药通道。
(3)心电监测。传统法无心电监护[21-23];改良组采用圆针穿过大鼠右上肢及双下肢指蹼,圆针于导联线相接(右上肢:绿色/右下肢:黑色/左下肢:红色),将导联线与医学信号采集处理系统连接,监测心电。
2.2.2 窒息诱导大鼠心脏骤停
手术完成后,记录基线信息(体温、心率、平均动脉压、心电图),适应性机械通气10 min(传统组:呼吸频率70次/min,潮气量0.7 mL/100 g;改良组:呼吸频率70次/min,潮气量2 mL/100 g),通气结束后关闭呼吸机,夹闭呼吸机管路诱导大鼠窒息。将开始窒息至平均动脉压降至 25 mmHg以下的时间以及心电图呈无脉性电活动或窦性停搏或室颤的时间定义为窒息致心脏停搏时间,间歇4 min后开始心肺复苏。
2.2.3 心肺复苏模型的建立
利用手动胸外按压和小动物呼吸机进行心肺复苏。在节拍器的引导下进行手动胸外按压,按压频率200次/min,深度为胸廓前后径的1/3;小动物呼吸机行机械通气,吸入氧浓度为 100%,呼吸频率70次/min,吸呼比1∶1,传统组潮气量0.7 mL/100 g,改良组潮气量2 mL/100 g;传统组推注肾上腺素0.01 mg,改良组推注肾上腺素0.02 mg,2 min重复注射一次,直至复苏成功或复苏失败。
2.2.4 复苏成功与失败的标准
平均动脉压≥60 mmHg(1 mmHg=133.22 Pa)且持续10 min的室上节律,被认定为复苏成功,记录复苏成功时间。心肺复苏进行10 min后自主循环仍未恢复,被认定为复苏失败。
2.2.5 复苏成功后处理
复苏成功后继续机械通气及持续监测心电图、血压、心率1 h,如果生命体征平稳则可停止机械通气和拔除各种导管,结扎血管,清理呼吸道分泌物,传统组直接放入鼠笼,改良组分3次皮下注射5%葡萄糖注射液共1 mL后放入鼠笼内,每8 h观察鼠笼情况,自由饮食饮水,室温控制在26 ℃。
2.3 观察指标
在复苏成功后24 h对大鼠意识、呼吸、颅神经反射、运动、感觉、协调进行神经功能损害评分(neurological function deficit scale,NDS),评分细则如表2所示,评分后采集大脑标本进行苏木精-伊红染色(hematoxylin-eosin,HE)观察神经细胞病理损伤情况,透射电镜观察神经元超微结构,Western Blot检测炎症、氧化应激、凋亡指标。
2.4 统计分析
所有实验均用至少3个生物学重复进行。在GraphPad Prism 9.5(GraphPad,CA,USA)中进行所有统计分析。两组间比较采用非配对t检验,两组以上比较采用单因素方差分析,数据用平均值±标准差(mean±SD)表示,*表示P<0.05,**表示P<0.01,差异具有统计学意义。
3 结果
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3.1 一般情况
术前大鼠反应敏捷,体毛光泽,饮食正常。共纳入93只大鼠,3只用作假手术组;传统组38只大鼠,复苏成功20只,复苏成功率52.63%(20/38);至复苏后24 h,存活12只,24 h存活率60%(12/20);改良组52只大鼠,复苏成功43只,复苏成功率82.69%(43/52);至复苏后24 h,存活36只,24 h存活率83.72%(36/43)。改良组复苏成功率和24 h存活率均高于传统组。
3.2 各组大鼠脑损伤情况
NDS评分显示,假手术组评分均为0分,传统组与改良组评分显著高于假手术组,如图4所示(P<0.05),传统组与改良组组间比较差异无统计学意义,如图4所示(P>0.05);HE染色显示,假手术组的脑组织正常,细胞排列致密均匀,无明显液泡间隙,核仁清晰可见,但传统组与改良组脑组织损伤较重,可见细胞排列紊乱,胞核固缩、裂解,深染,细胞间隙增宽,呈脑水肿表现,如图5所示。透射电镜显示,假手术组细胞结构正常,细胞核完整,传统组与改良组线粒体部分明显肿胀,膜完整,基质溶解,嵴减少、消失,可见空泡变,如图6所示。
3.3 各组大鼠炎症、氧化应激、凋亡指标情况
采用Western Blot检测各组大鼠脑组织中IL-6、SOD1、Bax、Bcl-2的蛋白表达,结果显示,与假手术组相比,传统组和改良组中IL-6、SOD1、Bax/Bcl-2表达显著增加;但传统组和改良组组间IL-6、SOD1、Bax/Bcl-2表达无显著差异,如图7所示(P<0.05)。
4 讨论
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由于临床研究的复杂性、不确定性及伦理学等原因,心脏骤停-心肺复苏动物模型一直是心肺复苏领域研究的基石[24]。自1996年Utstein-style Guidelines for Uniform Reporting of Laboratory CPR Research[25]在国际上发表以来,中外已发展出多种心脏骤停-心肺复苏模型,但是心肺复苏效果及神经功能障碍的研究仍未取得突破性的进展。由于全球人口的快速老龄化,心脏骤停的发生率和死亡率预计将会持续上升[26-28],因此,为探索良好的复苏效果和神经功能预后,稳定可靠的心脏骤停-心肺复苏动物模型一直都是医学领域关注的热点。
SD大鼠由于其可获得性和伦理学方面的优势,被广泛应用于医学研究,采用SD大鼠构心脏骤停-心肺复苏后脑损伤模型的常见方法为室颤法和窒息法。窒息法是较早并且长期使用的一种方式,常用于模拟临床上呼吸性原因导致的心脏骤停。研究表明,窒息法较室颤法脑缺血缺氧损伤更重,心脏骤停的复苏成功率更低,生存率也较低[29]。本课题组在前期预实验阶段也发现了这一问题,于是通过对传统窒息法的改良,建立了一种更高效的心脏骤停-心肺复苏后脑损伤模型,与传统法相比具有更高复苏成功率和24 h生存率,且传统组与改良组在脑损伤、炎症、氧化应激、凋亡等方面相当,能够较好地满足心肺复苏相关研究的需要。
首先,改良组复苏成功率达到82.69%,显著高于传统组的52.63%。这主要归因于气管插管、机械通气、胸外按压、肾上腺素的改进措施。
气管插管方面,传统法的气管插管主要采用间接照明经口直视下插管或者盲插法,上述方法的一次性插管成功率相对于可视喉镜下气管插管要低,多次气管插管可能会导致气道黏膜损伤、水肿、出血,增加了感染风险从而影响大鼠的生存率,且传统法气管插管后仅将导管固定在门齿,这种方法不仅在进行窒息诱导时和机械通气时容易滑脱,而且也难以保证气密性,从而影响窒息效果。改良组采用可视喉镜下气管插管,不仅提高了一次性插管的成功率,而且还保证了气道的通畅性;同时,在气管插管完成后将导管用1-0缝合线妥善固定于气管管壁,保障其气密性的同时还可以防止大鼠窒息和机械通气时导管滑脱。
机械通气方面,传统法主要在抢救期间采用潮气量0.7 mL/100 g,这种小潮气量的通气策略可能是基于肺保护性通气的考虑。2020年美国心脏协会在心肺复苏和心血管急救指南中建议[30],在心肺复苏抢救期间应给予足够的生理潮气量,潮气量要足以引起胸廓可见的起伏,但需避免过度通气,一旦恢复自主循环,应开始机械通气,强调了肺保护性通气策略,并根据患者的理想体重和血气分析结果调整潮气量;该指南并未在心肺复苏抢救期间建议行小潮气量的通气策略,而是在自主循环恢复后建议行小潮气量的保护性通气策略。小潮气量肺保护性通气策略主要适用于急性呼吸窘迫综合征患者、术后高危患者和长时间机械通气的ICU患者[31-32],这种策略旨在减少机械通气相关的肺损伤,提高患者的预后和生存率。但在心脏骤停的抢救情境下,保持较高的潮气量对确保有效的通气和氧合至关重要[33]。本课题组在前期预实验阶段发现心肺复苏抢救期间给予0.7 mL/100 g的小潮气量肺保护性通气策略其复苏成功率较低,可能与抢救期间通气和氧合不足有关,且本实验选择的SD大鼠为健康鼠,并无肺部的病变,这可能与小潮气量的保护性通气策略的适应症不符。后期改良后,给予其生理潮气量2 mL/100 g,复苏成功率较为满意,也印证了上述说法。
胸外按压方面,传统法主要采用人工手动胸外按压,其按压频率与深度较难统一,改良法虽然也是手动胸外按压,但是采用节拍器引导下的手动胸外按压,并且在按压的过程中观察心电监护与动脉血压监测,通过心电波形和压力波形来判断按压的有效性,确保了按压频率和深度的一致性和有效性。
肾上腺素方面,传统法的肾上腺素剂量一般是单次给予0.01 mg,2 min重复一次,在课题组多次试验探索中发现这种剂量下胸外按压时间较长,血压上升缓慢,复苏效果不佳,因此,将剂量调至0.02 mg之后,复苏时间及成功率较为满意,且复苏成功后血压维持情况相对较稳定。上述改进措施有效提高了心肺复苏的成功率。其次,改良组24 h生存率为83.72%,明显高于传统组的60%。高生存率不仅体现了复苏措施的有效性,还显示了术后管理的重要性。研究发现,SD大鼠在心肺复苏成功后的24 h,由于机体遭受心脏骤停的打击,会出现饮水进食以及活动的减少,从而影响其远期生存率[34]。本课题组在预实验阶段也观察到了这种现象,因此改良组在术后给予皮下注射5%葡萄糖注射液,提供了必要的营养支持,有助于动物的恢复。
尽管改良窒息法显示出明显的优势,但本次研究也存在一些局限性。如仅观察了复苏后24 h内的生存率和脑损伤情况,未能评估更长期的预后。此外,研究未对其他可能影响复苏效果的因素(如机械通气的氧浓度,大鼠的年龄、性别、健康状况等)进行详细分析。未来的研究应进一步优化模型,探索不同变量对复苏效果的影响,并评估长期预后和神经功能恢复情况。
5 结论
收起
改良窒息法通过标准化操作步骤、增加药物剂量、优化复苏参数和提供术后营养支持,提高了大鼠模型的复苏成功率和24 h生存率,并在脑损伤相当的前提下,改善了整体复苏效果。这一改良模型为研究全脑缺血再灌注损伤的病理机制和开发新疗法提供了一个可靠的动物模型,值得进一步的推广和应用。
基金
收起
  • 贵州省科技厅自然科学计划(黔科合基础-ZK【2023】一般401)
  • 贵州医科大学附属医院国家自然科学基金地区基金培育计划(gyfynsfc-2022-51)
  • 贵州医科大学附属医院博士科研启动基金(gyfybsky2021-65)
文献
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2025年第25卷第16期
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doi: 10.12404/j.issn.1671-1815.2406882
  • 接收时间：2024-09-13
  • 首发时间：2025-07-09
  • 出版时间：2025-06-08
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  • 收稿日期：2024-09-13
  • 修回日期：2025-03-24
基金
贵州省科技厅自然科学计划(黔科合基础-ZK【2023】一般401)
贵州医科大学附属医院国家自然科学基金地区基金培育计划(gyfynsfc-2022-51)
贵州医科大学附属医院博士科研启动基金(gyfybsky2021-65)
作者信息
    1 贵州医科大学临床医学院, 贵阳 550004
    2 贵州医科大学附属医院重症医学科, 贵阳 550004

通讯作者:

* 刘颖(1976—),女,汉族,贵州贵阳人,博士,主任医师。研究方向:脑缺血再灌注损伤机制。E-mail:。
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鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
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