芍药苷预处理对肝缺血再灌注大鼠肝功能及线粒体损伤的影响

目的探讨芍药苷预处理对肝缺血再灌注大鼠肝功能及线粒体损伤的影响。方法健康雄性SD大鼠64只,按照随机区组设计法分为假手术组(S组)、肝缺血再灌注组(I-R组)、芍药苷组(PF组)和生理盐水组(NS组),每组16只。I-R组、PF组和NS组采用肝脏缺血60min进行再灌注的方法制备70%肝脏缺血再灌注损伤模型。PF组于模型制备前7d通过尾静脉连续注射芍药苷30mg/(kg·d),共7d;NS组于模型制备前7d通过尾静脉连续注射0.9%Nacl注射液30mg/(kg·d),共7d。于再灌注6h后经心脏采集血液标本测定血清丙氨酸氨基转移酶(ALT)和天门冬氨酸氨基转移酶(AST)水平;取肝组织测定肝细胞凋亡情况,计算肝细胞凋亡指数(AI);制备肝细胞单细胞悬液,测定线粒体膜电位(△Ψm);提取肝组织线粒体,测定线粒体三态呼吸速率和四态呼吸速率(氧浓度降低值/实际时间),计算呼吸控制率(三态呼吸速率/四态呼吸速率)和磷氧比。结果与S组比较,I-R组和PF组大鼠AI、ALT、AST和四态呼吸速率水平明显升高,三态呼吸速率、呼吸控制率、磷氧比和△Ψm水平明显降低,差异有统计学意义(P<0.05);与I-R组比较,PF组大鼠AI、ALT、AST和四态呼吸速率水平明显降低,三态呼吸速率、呼吸控制率、磷氧比和△Ψm水平明显升高,差异有统计学意义(P<0.05)。结论芍药苷预处理可减轻大鼠肝缺血再灌注损伤,其机制可能与保护线粒体有关。

Effect of paeoniflorin pretreatment on damage of hepatic function and mitochondrion induced by hepatic ischemia-reperfusion in rats

Objective To investigate the effects of paeoniflorin pretreatment on hepatic function and mitochondrion induced by hepatic ischemia-reperfusion in rats.Methods A total of 64 adult SD rats were randomized divided into 4 groups,16 rats in each group,including sham operation group(group S),ischemia-reperfusion group(group I-R),paeoniflorin group(group PF)and normal saline group(group NS).The rats were anesthetized with intraperitoneal ketamine.A rat model of 70%hepatic ischemia-reperfusion injury was produced by occlusion of the liver for 60 min followed by 6 hreperfusion in group I-R,group PF and group NS.Group PF received resveratrol intravenously at 30 mg/(kg·d)for 7 days before modelling,while 0.9% Nacl 30 mg/(kg·d)was given in group NS.Blood samples were taken from the rat at 6 hof reperfusion for measurement of serum alanine aminotransferase(ALT)and aspartate aminotransferase(AST).The liver tissues were obtained to detect the liver cell apoptosis.Apoptotic index(AI)was calculated and the single liver cell suspension was prepared to determine△Ψm,three state respiration rate,four state respiration rate of mitochondria,respiratory control rate and P/O ratio.Results Compared with group S,the level of ALT,AST,AI and four state respiration rate in the group I-R and group PF increased,the level of three state respiration rate,respiratory control rate,P/O ratio and △Ψm decreased(P<0.05).Compared with group I-R,the level of ALT,AST,AI and four state respiration rate in the group PF decreased,the level of three state respiration rate,respiratory control rate,P/O ratio and△Ψm increased(P<0.05).Conclusion Paeoniflorin pretreatment could improve hepatic function induced by hepatic ischemia-reperfusion in rats,and protecting mitochondrion might be involved in the mechanism.