CAJ | 학술논문

目的评价辛二酰苯胺异羟肟酸(SAHA)对初进高原致死性失血性休克大鼠肝损伤的影响.方法选择健康成年雄性SD大鼠40只,体重240～280 g,2～3月龄,将大鼠从海拔1520 m养殖地运输至海拔3780m的实验基地,采用随机数字表法,将其分为4组(n=10):假手术组(S组)、致死性失血性休克组(LHS组)、生理盐水组(NS组)、SAHA组.LHS组、NS组和SAHA组经股动脉和股静脉放血制备致死性失血性休克模型.NS组和SAHA组失血完成后经静脉2min内分别注射0.25 ml生理盐水或SAHA 7.5 mg/kg(0.25 ml).记录大鼠3h生存情况.于失血前、失血完成后即刻、失血完成后3h时(存活时间不足3h时则于死亡即刻)采集股静脉血样,采用比色法检测血清AST、ALT、LDH活性.失于血完成后3h或死亡即刻取肝组织,采用Western blot法检测肝组织c-Jun氨基末端激酶(JNK)、磷酸化JNK(p-JNK)和caspase-3的表达和H3K9乙酰化水平.结果与S组比较,LHS组、NS组和SAHA组血清AST、ALT和LDH活性升高(P＜0.01).与LHS组和NS组比较,SAHA组血清AST、ALT和LDH活性降低,3h生存率升高,肝组织H3K9乙酰化水平升高,caspase-3表达下调,p-JNK/JNK降低(P＜ 0.05或0.01).LHS组和NS组肝组织病理学损伤严重,SAHA组病理学损伤程度明显减轻.结论初进高原致死性失血性休克大鼠休克早期应用SAHA具有肝保护作用,其作用机制与提高肝细胞H3K9乙酰化水平,抑制JNK/caspase-3凋亡信号转导通路有关.
목적 평개신이선분알이간우산(SAHA)대초진고원치사성실혈성휴극대서간손상적영향.방법 선택건강성년웅성SD대서40지,체중240～280 g,2～3월령,장대서종해발1520 m양식지운수지해발3780m적실험기지,채용수궤수자표법,장기분위4조(n=10):가수술조(S조)、치사성실혈성휴극조(LHS조)、생리염수조(NS조)、SAHA조.LHS조、NS조화SAHA조경고동맥화고정맥방혈제비치사성실혈성휴극모형.NS조화SAHA조실혈완성후경정맥2min내분별주사0.25 ml생리염수혹SAHA 7.5 mg/kg(0.25 ml).기록대서3h생존정황.우실혈전、실혈완성후즉각、실혈완성후3h시(존활시간불족3h시칙우사망즉각)채집고정맥혈양,채용비색법검측혈청AST、ALT、LDH활성.실우혈완성후3h혹사망즉각취간조직,채용Western blot법검측간조직c-Jun안기말단격매(JNK)、린산화JNK(p-JNK)화caspase-3적표체화H3K9을선화수평.결과 여S조비교,LHS조、NS조화SAHA조혈청AST、ALT화LDH활성승고(P＜0.01).여LHS조화NS조비교,SAHA조혈청AST、ALT화LDH활성강저,3h생존솔승고,간조직H3K9을선화수평승고,caspase-3표체하조,p-JNK/JNK강저(P＜ 0.05혹0.01).LHS조화NS조간조직병이학손상엄중,SAHA조병이학손상정도명현감경.결론 초진고원치사성실혈성휴극대서휴극조기응용SAHA구유간보호작용,기작용궤제여제고간세포H3K9을선화수평,억제JNK/caspase-3조망신호전도통로유관.
Objective To evaluate the effect of suberoylanilide hydroxamic acid (SAHA) on liver injury induced by lethal hemorrhagic shock in rats first entering high altitude.Methods Forty healthy male SpragueDawley rats,aged 2-3 months,weighing 240-280 g,transported from breeding grounds at an altitude of 1520 meters to the experimental station at an altitude of 3780 meters,were randomly divided into 4 groups (n =10each):sham operation group (group S),lethal hemorrhagic shock group (group LHS),normal saline group (group NS),and SAHA group.Anesthesia was induced with inhalation of 3% isoflurane and maintained with inhalation of 0.5%-1.0% isoflurane.Lethal hemorrhagic shock was induced by withdrawing blood from the femoral artery in groups LHS,NS and SAHA.Normal saline 0.25 ml and SAHA 7.5 mg/kg (0.25 ml) were injected intravenously over 2 min after completion of blood-letting in groups NS and SAHA,respectively.The survival rates with 3 h were recorded.Blood samples from femoral veins were taken before blood-letting,immediately after completion of blood-letting and at 3 h after completion of blood-letting (immediately after death if the survival time was less than 3 h) for determination of serum aspartate aminotransferase (AST),alanine aminotransferase (ALT) and lactic dehydrogenase (LDH) activities by the colorimetric method.Liver specimens were taken at 3 h after completion of blood-letting or immediately after death for examination of the pathological changes of the liver and for determination of c-Jun N-terminal kinase (JNK),phosphorylated-JNK (p-JNK) and caspase-3 expression and acetylation of H3K9 in liver tissues by Western blot.Results Compared with group S,the activities of serum AST,ALT and LDH were significantly increased in the other three groups (P ＜ 0.01).Compared with LHS and NS groups,the activities of serum AST,ALT and LDH were significantly decreased,the survival rate within 3 h and acetylation of H3K9 were increased,caspase-3 expression was down-regulated,and p-JNK/JNK ratio was decreased in group SAHA (P ＜ 0.05 or 0.01).The pathological changes of the liver were severe in LHS and NS groups and attenuated in SAHA group.Conclusion Administration of SAHA in early shock can significantly protect the liver after lethal hemorrhage in rats first entering high altitude,and increased acetylation of H3K9 and inhibition of the JNK/caspase-3 apoptotic pathway in liver tissues are involved in the mechanism.