中华麻醉学杂志
中華痳醉學雜誌
중화마취학잡지
CHINESE JOURNAL OF ANESTHESIOLOGY
2014年
1期
24-28
,共5页
二氧化碳%蛋白激酶C%再灌注损伤%肝
二氧化碳%蛋白激酶C%再灌註損傷%肝
이양화탄%단백격매C%재관주손상%간
Carbon dioxide%Protein kinase C%Reperfusion injury%Liver
目的 探讨蛋白激酶C(PKC)在CO2预处理减轻大鼠肝脏缺血再灌注损伤中的作用.方法 成年健康雄性Wistar大鼠48只,8～ 10周龄,体重230 ～ 270 g,采用随机数字表法分为3组(n=16):肝缺血再灌注损伤组(HIRI组)、CO2预处理组(P组)和PKC抑制剂白屈菜季铵碱组(CHE组).采用夹闭肝左中叶的门静脉、肝动脉以及胆管1h随后再灌注4h的方法建立肝缺血再灌注损伤模型.机械通气时HIRI组吸入50％ O2-50％ N2,P组吸入50％O2-45％ N2-5％ CO21h,随后吸入50％O2-50％N2,15 min后制备模型；CHE组机械通气前10 min腹腔注射CHE 5 mg/kg,其余操作同P组.于机械通气前、缺血前即刻、再灌注即刻、1h、2h、3h和4h时记录MAP,取动脉血样,行血气分析.再灌注4h时,测定血清ALT和AST活性,采用ELISA法测定血清TNF-α浓度,取肝组织,测定MDA含量和SOD活性(分光光度法)、活化的caspase-3(免疫组化法)和PKC(Western blot法)的表达水平,计算凋亡指数(TUNEL法).结果 与HIRI组比较,P组缺血前即刻和再灌注期间MAP、PaO2和PaCO2升高,CHE组再灌注期间MAP和PaCO2升高,缺血前即刻和再灌注期间PaO2升高,P组和CHE组血清ALT和AST活性、TNF-α浓度、肝组织MDA含量降低,细胞凋亡指数降低,肝组织活化的caspase-3表达下调,P组SOD活性升高,PKC表达上调(P＜0.05或0.01),CHE组SOD活性和PKC表达水平差异无统计学意义(P＞0.05)；与P组比较,CHE组MAP再灌注即刻升高,再灌注1-4 h时降低,缺血前即刻和再灌注期间PaO2降低,再灌注3h时PaCO2降低,血清ALT和AST活性、TNF-α浓度、肝组织MDA含量和细胞凋亡指数升高,肝组织活化的caspase-3表达上调,PKC表达下调(P＜0.05).结论 PKC参与了CO2预处理减轻大鼠肝缺血再灌注损伤的作用.
目的 探討蛋白激酶C(PKC)在CO2預處理減輕大鼠肝髒缺血再灌註損傷中的作用.方法 成年健康雄性Wistar大鼠48隻,8～ 10週齡,體重230 ～ 270 g,採用隨機數字錶法分為3組(n=16):肝缺血再灌註損傷組(HIRI組)、CO2預處理組(P組)和PKC抑製劑白屈菜季銨堿組(CHE組).採用夾閉肝左中葉的門靜脈、肝動脈以及膽管1h隨後再灌註4h的方法建立肝缺血再灌註損傷模型.機械通氣時HIRI組吸入50％ O2-50％ N2,P組吸入50％O2-45％ N2-5％ CO21h,隨後吸入50％O2-50％N2,15 min後製備模型；CHE組機械通氣前10 min腹腔註射CHE 5 mg/kg,其餘操作同P組.于機械通氣前、缺血前即刻、再灌註即刻、1h、2h、3h和4h時記錄MAP,取動脈血樣,行血氣分析.再灌註4h時,測定血清ALT和AST活性,採用ELISA法測定血清TNF-α濃度,取肝組織,測定MDA含量和SOD活性(分光光度法)、活化的caspase-3(免疫組化法)和PKC(Western blot法)的錶達水平,計算凋亡指數(TUNEL法).結果 與HIRI組比較,P組缺血前即刻和再灌註期間MAP、PaO2和PaCO2升高,CHE組再灌註期間MAP和PaCO2升高,缺血前即刻和再灌註期間PaO2升高,P組和CHE組血清ALT和AST活性、TNF-α濃度、肝組織MDA含量降低,細胞凋亡指數降低,肝組織活化的caspase-3錶達下調,P組SOD活性升高,PKC錶達上調(P＜0.05或0.01),CHE組SOD活性和PKC錶達水平差異無統計學意義(P＞0.05)；與P組比較,CHE組MAP再灌註即刻升高,再灌註1-4 h時降低,缺血前即刻和再灌註期間PaO2降低,再灌註3h時PaCO2降低,血清ALT和AST活性、TNF-α濃度、肝組織MDA含量和細胞凋亡指數升高,肝組織活化的caspase-3錶達上調,PKC錶達下調(P＜0.05).結論 PKC參與瞭CO2預處理減輕大鼠肝缺血再灌註損傷的作用.
목적 탐토단백격매C(PKC)재CO2예처리감경대서간장결혈재관주손상중적작용.방법 성년건강웅성Wistar대서48지,8～ 10주령,체중230 ～ 270 g,채용수궤수자표법분위3조(n=16):간결혈재관주손상조(HIRI조)、CO2예처리조(P조)화PKC억제제백굴채계안감조(CHE조).채용협폐간좌중협적문정맥、간동맥이급담관1h수후재관주4h적방법건립간결혈재관주손상모형.궤계통기시HIRI조흡입50％ O2-50％ N2,P조흡입50％O2-45％ N2-5％ CO21h,수후흡입50％O2-50％N2,15 min후제비모형；CHE조궤계통기전10 min복강주사CHE 5 mg/kg,기여조작동P조.우궤계통기전、결혈전즉각、재관주즉각、1h、2h、3h화4h시기록MAP,취동맥혈양,행혈기분석.재관주4h시,측정혈청ALT화AST활성,채용ELISA법측정혈청TNF-α농도,취간조직,측정MDA함량화SOD활성(분광광도법)、활화적caspase-3(면역조화법)화PKC(Western blot법)적표체수평,계산조망지수(TUNEL법).결과 여HIRI조비교,P조결혈전즉각화재관주기간MAP、PaO2화PaCO2승고,CHE조재관주기간MAP화PaCO2승고,결혈전즉각화재관주기간PaO2승고,P조화CHE조혈청ALT화AST활성、TNF-α농도、간조직MDA함량강저,세포조망지수강저,간조직활화적caspase-3표체하조,P조SOD활성승고,PKC표체상조(P＜0.05혹0.01),CHE조SOD활성화PKC표체수평차이무통계학의의(P＞0.05)；여P조비교,CHE조MAP재관주즉각승고,재관주1-4 h시강저,결혈전즉각화재관주기간PaO2강저,재관주3h시PaCO2강저,혈청ALT화AST활성、TNF-α농도、간조직MDA함량화세포조망지수승고,간조직활화적caspase-3표체상조,PKC표체하조(P＜0.05).결론 PKC삼여료CO2예처리감경대서간결혈재관주손상적작용.
Objective To investigate the role of protein kinase C (PKC) in reduction of hepatic ischemiareperfusion injury by CO2 preconditioning in rats.Methods Forty-eight male Wistar rats,aged 8-10 weeks,weighing 230-270 g,were randomly divided into 3 groups (n =16 each):hepatic ischemia-reperfusion injury group (group HIRI),CO2 preconditioning group (group P),and c helerythrine (CHE,a specific inhibitor of PKC) group (group CHE).The portal vein,hepatic artery and bile duct of the left lateral and median lobes of the liver were occluded for 1 h,followed by 4 h reperfusion in anesthetized rats.The rats inhaled 50％ O2-50％ N2 for 1 h during mechanical ventilation in group HIRI.In P group,the rats inhaled 50％ O2-45％ N2-5％ CO2 for 1 h during mechanical ventilation and then inhaled 50％ O2-50％ N2 and the hepatic ischemia-reperfusion injury was performed 15 min later.In group CHE,CHE 5 mg/kg was injected intraperitoneally at 10 min before mechanical ventilation,and the other procedures were similar to those previously described in P group.Before mechanical ventilation,immediately before ischemia,and at 0,1,2,3 and 4 h of reperfusion,mean arterial pressure (MAP) was recorded and arterial blood samples were obtained for blood gas analysis.At 4 h of reperfusion,the serum aspartate amino transferase (AST) and alanine amino-transferase (ALT) activities and tumor necrosis factor-α (TNF-α) concentration (by ELISA) were determined and hepatic specimens were obtained for detection of malondialdehyde (MDA) content and superoxide dismutase (SOD) activity (by spectrophotometry),and the expression of activated caspase-3 (by immuno-histochemistry) and PKC (by Western blot) in hepatic tissues.Apoptosis index was calculated by using TUNEL.Results Compared with group HIRI,MAP,PaO2 and PaCO2were significantly increased immediately before ischemia and during reperfusion in group P,MAP and PaCO2 were increased during reperfusion and PaO2 was increased immediately before ischemia and during reperfusion in group CHE,the serum ALT and AST activities,TNF-α concentrations,MDA content and apoptosis index were decreased,and the expression of activated caspase-3 was down-regulated in P and CHE groups,and the SOD activity was increased,and the expression of PKC was up-regulated in group P (P ＜ 0.05 or 0.01),and no significant changes were found in the SOD activity and PKC expression in CHE group (P ＞ 0.05).Compared with group P,MAP was significantly increased immediately after onset of reperfusion,while decreased at 1-4 h of reperfusion,PaO2 was decreased immediately before ischemia and during reperfusion,PaCO2 was decreased at 3 h of reperfusion,the serum ALT and AST activities,TNF-α concentrations,MDA content and apoptosis index were increased,and the expression of activated caspase-3 was up-regulated,and the expression of PKC was downregulated in group CHE (P ＜ 0.05).Conclusion PKC is involved in reduction of hepatic ischemia-reperfusion injury by CO2 preconditioning in rats.
