CAJ | 학술논문

目的建立胞壁酰二肽(MDP)诱发烫伤大鼠脓毒症模型.方法选择SPF级健康雄性SD大鼠50只,体重200～250 g,2～3月龄,采用随机数字表法,将其分为3组:对照组(C组,n=10)、烫伤组(S组,n=10),制备20%总体表面积Ⅲ度烫伤模型；MDP组(n=30),于烫伤后24h时经股静脉注射MDP 5 mg/kg.MDP组于注射MDP后1、6、24h时,S组于烫伤后24h时,C组于20℃水中假烫伤后24h时取腹主动脉血样,分别行血气分析、计数WBC和Plt、检测肝肾功能、肌酸激酶同工酶(CK-MB)活性、血浆TNF-α、干扰素-γ(IFN-γ)、IL-6、IL-10、高迁移率族蛋白-1(HMGB-1)水平；取血样后处死大鼠,取心、肝、肺和肾组织行病理学检查,另取肺组织测定髓过氧化物酶(MPO)活性.另取雄性SD大鼠90只,按照上述方法分组处理后,记录72 h生存情况.结果与C组比较,S组血浆IL-6、IL-10、IFN-γ和HMGB1水平、WBC计数、血清ALT、AST和BUN水平、肺组织MPO活性升高,生存率降低,MDP组血浆TNF-α、IL-6、IL-10、IFN-γ和HMGB-1水平、血清ALT、AST、总胆红素、BUN、Cr和CK-MB水平、肺组织MPO活性、PaCO2升高,BE负值增加,WBC和Plt计数、pH值、PaO2和生存率降低(P＜0.05).与S组比较,MDP组血浆TNF-α、IL-6、IFN-γ和HMGB-1水平,血清ALT、AST、总胆红素、BUN、Cr、CK-MB水平、肺组织MPO活性和PaC02升高,BE负值增加,WBC和Plt计数、pH值、PaO2和生存率降低(P＜0.05).S组和MDP组心、肝、肺和肾组织病理学损伤明显,MDP组病理学损伤程度较重.结论 Ⅲ度烫伤后经MDP诱导可致大鼠广泛炎性反应并伴多器官功能损伤,成功建立烫伤后脓毒症模型.
목적 건립포벽선이태(MDP)유발탕상대서농독증모형.방법 선택SPF급건강웅성SD대서50지,체중200～250 g,2～3월령,채용수궤수자표법,장기분위3조:대조조(C조,n=10)、탕상조(S조,n=10),제비20%총체표면적Ⅲ도탕상모형；MDP조(n=30),우탕상후24h시경고정맥주사MDP 5 mg/kg.MDP조우주사MDP후1、6、24h시,S조우탕상후24h시,C조우20℃수중가탕상후24h시취복주동맥혈양,분별행혈기분석、계수WBC화Plt、검측간신공능、기산격매동공매(CK-MB)활성、혈장TNF-α、간우소-γ(IFN-γ)、IL-6、IL-10、고천이솔족단백-1(HMGB-1)수평；취혈양후처사대서,취심、간、폐화신조직행병이학검사,령취폐조직측정수과양화물매(MPO)활성.령취웅성SD대서90지,안조상술방법분조처리후,기록72 h생존정황.결과 여C조비교,S조혈장IL-6、IL-10、IFN-γ화HMGB1수평、WBC계수、혈청ALT、AST화BUN수평、폐조직MPO활성승고,생존솔강저,MDP조혈장TNF-α、IL-6、IL-10、IFN-γ화HMGB-1수평、혈청ALT、AST、총담홍소、BUN、Cr화CK-MB수평、폐조직MPO활성、PaCO2승고,BE부치증가,WBC화Plt계수、pH치、PaO2화생존솔강저(P＜0.05).여S조비교,MDP조혈장TNF-α、IL-6、IFN-γ화HMGB-1수평,혈청ALT、AST、총담홍소、BUN、Cr、CK-MB수평、폐조직MPO활성화PaC02승고,BE부치증가,WBC화Plt계수、pH치、PaO2화생존솔강저(P＜0.05).S조화MDP조심、간、폐화신조직병이학손상명현,MDP조병이학손상정도교중.결론 Ⅲ도탕상후경MDP유도가치대서엄범염성반응병반다기관공능손상,성공건립탕상후농독증모형.
Objective To establish a rat model of sepsis induced by muramyl dipeptide (MDP) after scald burn.Methods Fifty SPF male Sprague-Dawley rats,aged 2-3 months,weighing 200-250 g,were randomly divided into 3 groups:control group (group C,n =10),scald group (group S,n =10) and MDP group (n =30).The rats were subjected to a third-degree scald burn covering 20% of total body surface area in groups S and MDP.The rats were only exposed to 20 ℃ water in group C.MDP 5 mg/kg was injected via the femoral vein at 24 h after scald bum in group MDP.Arterial blood samples were collected at 1,6 and 24 h after MDP injection in group MDP,at 24 h after scald burn in group S,or at 24 h after exposure to 20 ℃ water in group C for blood gas analysis and for measurement of white blood cell (WBC) and platelet (Plt) counts,serum aminotransferase (ALT),aspartate transferase (AST),total bilirubin (TB),creatinine (Cr) and blood urea nitrogen (BUN) levels,creatine kinase isoenzyme-MB (CK-MB) activity,and plasma tumor necrosis factor-α (TNF-α),interferon-γ(IFN-γ),interleukin-6 (IL-6),IL-10 and high mobility group box 1 protein (HMGB-1) levels.The rats were sacrificed after collecting blood samples,and heart,liver,lung,and kidney specimens were obtained for microscopic examination of pathologic changes.The activity of myeloperoxidase (MPO) in lung tissues was measured.Another 90 male Sprague-Dawley rats were randomly divided into 3 groups and treated as the method previously described for record of the survival rate within 72 h.Results Compared with C group,the plasma IL-6,IL-10,IFN-γ and HMGB1 levels,WBC count,serum ALT,AST,and BUN levels and MPO activity were significantly increased,and the survival rate within 72 h was decreased in S group,and the plasma TNF-α,IL-6,IL-10,IFN-γand HMGB-1 levels,serum ALT,AST,TB,BUN,Cr and CK-MB levels,MPO activity,PaCO2 and BE value were significantly increased,and WBC and PLT counts,pH value,PaO2 and survival rate within 72 h were decreased in MDP group (P ＜ 0.05).Compared with S group,the plasma TNF-α,IL-6,IFN-γ and HMGB-1 levels,serum ALT,AST,TB,BUN,Cr and CK-MB levels,MPO activity,PaCO2 and BE value were significantly increased,and WBC and Plt counts,pH value,PaO2 and survival rate within 72 h were decreased in MDP group (P ＜ 0.05).The pathologic changes of heart,liver,lung and kidney were obvious in S and MDP groups and severer in MDP group.Conclusion After a third-degree 20% total body surface area scald burn,MDP induces excessive production of inflammatory cytokines accompanying with multiple organ damage ; thus the model of sepsis is successfully established after scald burn in rats.