摘要
目的明确4种肝缺血/再灌注损伤(IRI)大鼠模型的制备方法,确定与临床情况相符、病理生理损伤程度稳定、易于操作的肝IRI动物模型。方法将160只雄性SD大鼠采用区间分组法随机分为70%IRI(A组)、100%IRI(B组)、70%IRI+30%肝切除(C组)、100%IRI+30%肝切除(D组)4种模型组,每组40只;每种模型再按缺血时间随机分为假手术组(s组)和缺血30、60、90 min组,每组10只。术后观察大鼠生存状态、苏醒时间;记录C、D组切除肝叶质量、出血量、止血时间。于再灌注6 h经心脏穿刺取血,检测血清天冬氨酸转氨酶(AST)、丙氨酸转氨酶(ALT)、碱性磷酸酶(ALP)、血尿素氮(BUN)、血肌酐(SCr)、γ-谷氨酰转肽酶(γ-GT)水平,判断肝肾功能损伤情况;采用苏木素-伊红(HE)染色、免疫组化巨噬细胞染色,从病理学角度分析肝组织结构损伤情况。结果A组大鼠苏醒早,精神尚可;余组大鼠苏醒晚且精神欠佳。D组止血时间平均较C组长1 s左右。70%肝缺血60 min组大鼠的死亡率为0。与假手术组比较,各实验组大鼠血清AST、ALT、ALP、BUN、SCr、γ-GT水平明显增高,提示肝IRI大鼠肝肾功能受损;A、B、C 3组中缺血90 min组较缺血30 min组AST、ALT、ALP、BUN、SCr、γ-GT升高更明显〔AST(U/L):A组为834.94±56.73比258.74±18.33,B组为547.63±217.40比277.67±57.92,C组为930.38±75.48比640.51±194.20;ALT(U/L):A组为346.78±25.47比156.58±13.25,B组为408.40±138.25比196.80±58.60,C组为596.41±193.32比173.76±72.43,ALP(U/L):A组为431.21±34.30比315.95±15.64,B组为525.88±62.13比215.63±17.31,C组为487.53±112.37比272.46±92.33,BUN(U/L):A组为18.35±5.63比14.32±2.30,B组为30.21±4.55比17.41±8.14,C组为20.50±3.64比15.93±3.22,SCr(U/L):A组为27.47±8.91比22.37±5.66,B组为43.60±15.57比36.80±7.95,C组为63.81±20.24比42.47±7.03,γ-GT(U/L):A组为15.64±3.57比6.82±1.48,B组为9.28±1.91比5.62±1.21,C组为10.98±3.18比5.67±1.10,均P<0.05〕;100%IRI 90 min组和100%IRI 90
Objective To clarify the preparation methods of four rat models of liver ischemia/reperfusion injury(IRI)and to determine a liver IRI animal model that is consistent with clinical conditions,has stable pathological and physiological injury,and is easy to operate.Methods A total of 160 male Sprague-Dawley(SD)rats were randomly divided into four groups using an interval grouping method:70%IRI(group A),100%IRI(group B),70%IRI with 30%hepatectomy(group C),and 100%IRI with 30%hepatectomy(group D),with 40 rats in each group.Each model was further divided into sham operation group(S group)and ischemia groups of 30,60,and 90 minutes,with 10 rats in each group.After surgery,the survival status and awakening time of the rats were observed,and the liver lobectomy weight,bleeding volume,and hemostasis time of groups C and D were recorded.Blood samples were collected by cardiac puncture after 6 hours of reperfusion for determination the levels of aspartate aminotransferase(AST),alanine aminotransferase(ALT),alkaline phosphatase(ALP),blood urea nitrogen(BUN),serum creatinine(SCr),andγ-glutamyl transpeptidase(γ-GT)in the serum to assess liver and kidney function.Hematoxylin-eosin(HE)staining and immunohistochemical staining of macrophages were performed to analyze the liver tissue structure damage from a pathological perspective.Results Rats in group A exhibited earlier awakening and acceptable mental status,while rats in the other groups showed delayed awakening and poor mental status.The hemostasis time in group D was approximately 1 second longer than that in group C.The mortality of rats subjected to 60 minutes of 70%hepatic ischemia was 0.Compared to the sham operation group,rats in each experimental group showed significant increases in serum levels of AST,ALT,ALP,BUN,SCr,andγ-GT,indicating impaired liver and kidney function in the rat models of liver IRI.In groups A,B,and C,the 90-minute ischemia subgroup exhibited more pronounced elevation in AST,ALT,ALP,BUN,SCr,andγ-GT levels compared to the 30-minute ischemia subg
作者
罗嘉琪
王莉荔
陈阜东
张爱舷
张晗
张晓萌
陈力
Luo Jiaqi;Wang Lili;Chen Fudong;Zhang Aixian;Zhang Han;Zhang Xiaomeng;Chen Li(Academy of Military Medical Sciences,Beijing 100853,China;Department of General Practice,First Medical Center,General Hospital of the Chinese People's Liberation Army,Beijing 100853,China)
出处
《中华危重病急救医学》
CAS
CSCD
北大核心
2023年第6期604-609,共6页
Chinese Critical Care Medicine
基金
军队基础加强计划技术领域基金项目(2019-JCJQ-JJ-143)。
关键词
大鼠
急性肝缺血/再灌注损伤
模型制备
适用性
Rat
Acute liver ischemia/reperfusion injury
Model preparation
Applicability
