目的 评价限制性液体复苏对创伤失血性休克大鼠全身炎性反应的影响.方法 健康清洁级雄性SD大鼠60只,2～3月龄,体重250 ～ 290 g,采用随机数字表法,将其分为6组(n=10):假手术组(S组)、无液体复苏组(NF组)、非限制性液体复苏组(ULF组)、限制性晶体液复苏组(LR组)、不同限制性胶体液复苏组(LSG组和LHES组).采用放血+断尾法制备创伤未控制失血性休克模型.股动脉20 min内放血2.5 ml/100 g,放血结束后10 min断尾,同时行液体复苏,分别静脉输注乳酸钠林格氏液(ULF组和LR组)、4％琥珀酰明胶(LSG组)及6％羟乙基淀粉130/0.4(LHES组).初始输注速率为2 ml·kg-1·min-1,限制性液体复苏目标MAP 50 mm Hg,非限制性液体复苏目标MAP 80 mm Hg.60 min后尾部断端止血并停止液体输注,改用血液复苏60 min后进入观察期180 min.于股动、静脉置管术后稳定10 min(T0)、股动脉放血结束后10 min(T1)、液体复苏结束(T2)、血液复苏结束(T3)及观察期结束(T4)时采集股动脉血样,测定Hct、血浆TNF-α、IL-6及IL-10的浓度；于T2时采集股动脉血样,检测单核细胞Toll样受体4(TLR4)及髓样细胞分化蛋白88(MyD88)表达水平和NF-κB活性；记录尾部断端出血量及输液量.另取SD大鼠120只,按照上述方法分组(n=20)和复苏处理后,观察72 h内生存情况.结果 与S组比较,其余各组Hct降低,血浆TNF-α、IL-6、IL-10浓度升高,单核细胞TLR4及MyD88表达上调,NF-κB活性增强(P＜0.05).与NF组比较,ULF组、LR组及LSG组血浆TNF-α、IL-6浓度升高,IL-10浓度和Hct降低,单核细胞TLR4及MyD88表达上调,NF-κB活性增强,LHES组血浆TNF-α、IL-6浓度升高,IL-10浓度和Hct降低(P＜0.05).与ULF组比较,LSG组、LHES组及LR组血浆TNF-α、IL-6浓度降低,IL-10浓度、Hct和生存率升高,单核细胞TLR4及MyD88表达下调,NF-κB活性减弱,尾部断端出血量降低,输液量减少(P＜0.05).与LR组比较,LHES组血浆TNF-α、IL-6浓度降低,单核细胞TLR4及MyD88表达下调,NF-κB活性减弱(P＜0.05),IL-10浓度差异无统计学意义(P＞0.05),LSG组及LHES组输液量减少,生存率升高(P＜0.05),尾部断端出血量比较差异无统计学意义(P＞0.05).结论 与非限制性液体复苏比较,限制性液体复苏对创伤失血性休克大鼠全身炎性反应影响小,且6％羟乙基淀粉130/0.4复苏效果尤其显著,其机制与抑制TLR4/NF-κB信号通路有关.
目的 評價限製性液體複囌對創傷失血性休剋大鼠全身炎性反應的影響.方法 健康清潔級雄性SD大鼠60隻,2～3月齡,體重250 ～ 290 g,採用隨機數字錶法,將其分為6組(n=10):假手術組(S組)、無液體複囌組(NF組)、非限製性液體複囌組(ULF組)、限製性晶體液複囌組(LR組)、不同限製性膠體液複囌組(LSG組和LHES組).採用放血+斷尾法製備創傷未控製失血性休剋模型.股動脈20 min內放血2.5 ml/100 g,放血結束後10 min斷尾,同時行液體複囌,分彆靜脈輸註乳痠鈉林格氏液(ULF組和LR組)、4％琥珀酰明膠(LSG組)及6％羥乙基澱粉130/0.4(LHES組).初始輸註速率為2 ml·kg-1·min-1,限製性液體複囌目標MAP 50 mm Hg,非限製性液體複囌目標MAP 80 mm Hg.60 min後尾部斷耑止血併停止液體輸註,改用血液複囌60 min後進入觀察期180 min.于股動、靜脈置管術後穩定10 min(T0)、股動脈放血結束後10 min(T1)、液體複囌結束(T2)、血液複囌結束(T3)及觀察期結束(T4)時採集股動脈血樣,測定Hct、血漿TNF-α、IL-6及IL-10的濃度；于T2時採集股動脈血樣,檢測單覈細胞Toll樣受體4(TLR4)及髓樣細胞分化蛋白88(MyD88)錶達水平和NF-κB活性；記錄尾部斷耑齣血量及輸液量.另取SD大鼠120隻,按照上述方法分組(n=20)和複囌處理後,觀察72 h內生存情況.結果 與S組比較,其餘各組Hct降低,血漿TNF-α、IL-6、IL-10濃度升高,單覈細胞TLR4及MyD88錶達上調,NF-κB活性增彊(P＜0.05).與NF組比較,ULF組、LR組及LSG組血漿TNF-α、IL-6濃度升高,IL-10濃度和Hct降低,單覈細胞TLR4及MyD88錶達上調,NF-κB活性增彊,LHES組血漿TNF-α、IL-6濃度升高,IL-10濃度和Hct降低(P＜0.05).與ULF組比較,LSG組、LHES組及LR組血漿TNF-α、IL-6濃度降低,IL-10濃度、Hct和生存率升高,單覈細胞TLR4及MyD88錶達下調,NF-κB活性減弱,尾部斷耑齣血量降低,輸液量減少(P＜0.05).與LR組比較,LHES組血漿TNF-α、IL-6濃度降低,單覈細胞TLR4及MyD88錶達下調,NF-κB活性減弱(P＜0.05),IL-10濃度差異無統計學意義(P＞0.05),LSG組及LHES組輸液量減少,生存率升高(P＜0.05),尾部斷耑齣血量比較差異無統計學意義(P＞0.05).結論 與非限製性液體複囌比較,限製性液體複囌對創傷失血性休剋大鼠全身炎性反應影響小,且6％羥乙基澱粉130/0.4複囌效果尤其顯著,其機製與抑製TLR4/NF-κB信號通路有關.
목적 평개한제성액체복소대창상실혈성휴극대서전신염성반응적영향.방법 건강청길급웅성SD대서60지,2～3월령,체중250 ～ 290 g,채용수궤수자표법,장기분위6조(n=10):가수술조(S조)、무액체복소조(NF조)、비한제성액체복소조(ULF조)、한제성정체액복소조(LR조)、불동한제성효체액복소조(LSG조화LHES조).채용방혈+단미법제비창상미공제실혈성휴극모형.고동맥20 min내방혈2.5 ml/100 g,방혈결속후10 min단미,동시행액체복소,분별정맥수주유산납림격씨액(ULF조화LR조)、4％호박선명효(LSG조)급6％간을기정분130/0.4(LHES조).초시수주속솔위2 ml·kg-1·min-1,한제성액체복소목표MAP 50 mm Hg,비한제성액체복소목표MAP 80 mm Hg.60 min후미부단단지혈병정지액체수주,개용혈액복소60 min후진입관찰기180 min.우고동、정맥치관술후은정10 min(T0)、고동맥방혈결속후10 min(T1)、액체복소결속(T2)、혈액복소결속(T3)급관찰기결속(T4)시채집고동맥혈양,측정Hct、혈장TNF-α、IL-6급IL-10적농도；우T2시채집고동맥혈양,검측단핵세포Toll양수체4(TLR4)급수양세포분화단백88(MyD88)표체수평화NF-κB활성；기록미부단단출혈량급수액량.령취SD대서120지,안조상술방법분조(n=20)화복소처리후,관찰72 h내생존정황.결과 여S조비교,기여각조Hct강저,혈장TNF-α、IL-6、IL-10농도승고,단핵세포TLR4급MyD88표체상조,NF-κB활성증강(P＜0.05).여NF조비교,ULF조、LR조급LSG조혈장TNF-α、IL-6농도승고,IL-10농도화Hct강저,단핵세포TLR4급MyD88표체상조,NF-κB활성증강,LHES조혈장TNF-α、IL-6농도승고,IL-10농도화Hct강저(P＜0.05).여ULF조비교,LSG조、LHES조급LR조혈장TNF-α、IL-6농도강저,IL-10농도、Hct화생존솔승고,단핵세포TLR4급MyD88표체하조,NF-κB활성감약,미부단단출혈량강저,수액량감소(P＜0.05).여LR조비교,LHES조혈장TNF-α、IL-6농도강저,단핵세포TLR4급MyD88표체하조,NF-κB활성감약(P＜0.05),IL-10농도차이무통계학의의(P＞0.05),LSG조급LHES조수액량감소,생존솔승고(P＜0.05),미부단단출혈량비교차이무통계학의의(P＞0.05).결론 여비한제성액체복소비교,한제성액체복소대창상실혈성휴극대서전신염성반응영향소,차6％간을기정분130/0.4복소효과우기현저,기궤제여억제TLR4/NF-κB신호통로유관.
Objective To evaluate the effects of limited fluid resuscitation on systemic inflammatory responses in rats with traumatic hemorrhagic shock through comparing with unlimited fluid resuscitation.Methods Sixty pathogen-free male Sprague-Dawley rats,aged 2-3 months,weighing 250-290 g,were randomly divided into 6 groups (n =10 each) using a random number table:sham operation group (group S),no fluid resuscitation group (group NF),unlimited fluid resuscitation group (group ULF),limited crystalloid fluid resuscitation group (group LR),and limited colloid fluid resuscitation groups (group LSG and group LHES).Traumatic uncontrolled hemorrhagic shock was induced by withdrawal of blood from the femoral artery at 2.5 mL/100 g over a 20-minute period,followed by tail amputation at 10 min after the end of blood withdrawal.At 10 min after the end of blood withdrawal,fluid resuscitation was performed.Lactated Ringer's solution (ULF and LR groups),4 ％ succinylated gelatin (group LSG),or 6 ％ hydroxyethyl starch 130/0.4 (group LHES) was infused intravenously.The initial infusion rate was 2 ml · kg-1 · min-1.The target MAP was maintained at 50 mm Hg in rats with limited fluid resuscitation,while at 80 mm Hg in rats with unlimited fluid resuscitation.After 60 min of fluid resuscitation,bleeding in the tail was stopped by ligation and fluid infusion was replaced with blood resuscitation.After 60 min of blood resuscitation,180 main of observation was started.At 10 min after catheterization of the femoral artery and vein (T0),10 min after the end of blood withdrawal (T1),the end of fluid resuscitation (T2),the end of blood resuscitation (T3),and the end of observation (T4),arterial blood samples were collected to measure hematocrit (Hct)and concentrations of plasma tumor necrosis factor-alpha (TNF-α),interleukin (IL)-6,and IL-10.Blood samples were collected from the femoral artery at T2 for determination of the expression of Toll-like receptor 4 (TLR4) and myeloid differentiation factor 88 (MyD88) and activity of nuclear factor-kappaB (NF-κB) in monocytes.The amount of blood loss from the tail and volume of fluid infused were also recorded.Another 120 Sprague-Dawley rats were randomly divided into 6 groups (n =20 each) and resuscitation was performed according to the method previously described.The rats were observed for 72 h survival rate.Results Compared with group S,Hct was significantly decreased,the concentrations of plasma TNF-α,IL-6,and IL-10 and activity of NF-κB were increased,and the expression of TLR4,and MyD88 in monocytes was up-regulated in the other groups (P ＜ 0.05).Compared with group NF,the concentrations of plasma TNF-α and IL-6 and NF-κB activity were significantly increased,and the concentration of plasma IL-10 and Hct were decreased,and the expression of TLR4 and MyD88 in monocytes was up-regulated in ULF,LR and LSG groups,and the concentrations of plasma TNF-α and IL-6 were significantly increased,the concentration of plasma IL-10 and Hct were decreased in group LHES (P ＜ 0.05).Compared with group ULF,the concentrations of plasma TNF-α and IL-6 and NF-κB activity were significantly decreased,the concentration of plasma IL-10 and Hct were increased,the survival rate was higher,the expression of TLR4 and MyD88 in monocytes was down-regulated,and the amount of blood loss from the tail was decreased and the volume of fluid infused was reduced in LSG,LHES and LR groups (P ＜ 0.05).Compared with group LR,the concentrations of plasma TNF-α and IL-6 and NF-κB activity were significantly decreased and the expression of TLR4 and MyD88 in monocytes was down-regulated (P ＜ 0.05),and no significant change was found in the concentration of plasma IL-10 in group LHES (P ＞ 0.05),and the volume of fluid infused was reduced and the survival rate was increased (P ＜ 0.05),and no significant change was found in the amount of blood loss from the tail in LSG and LH-ES groups (P ＞ 0.05).Conclusion Compared with unlimited fluid resuscitation,limited fluid resuscitation exerts less effect on systemic inflammatory responses in rats with traumatic hemorrhagic shock,especially when resuscitation with 6％ hydroxyethyl starch 130/0.4 is performed,and inhibition of TLR4/NF-κB signaling pathway is involved in the mechanism.
