1,6-二磷酸果糖与环孢素A对心脏移植大鼠心室肌细胞凋亡的协同作用

Synergistic action of fructose-1,6-diphosphate combined with cyclosporine A in the apoptosis of ventricular myocytes following cardiac allograft in rats

目的:观察1,6-二磷酸果糖与环孢素A单独与联合应用对同种异体大鼠心脏移植后急性排斥期心肌细胞凋亡的影响。方法:实验于2004-04/12在南京军区南京总医院动物实验中心完成。选择Wistar大鼠(供体)和SD大鼠(受体)各40只,根据Ono改进模型将Wistar大鼠心脏移植到SD大鼠腹腔,随机数字表法分为4组,各组10例。对照组,手术前后不作处理;1,6-二磷酸果糖组,供体术前5d开始每天静脉注射1,6-二磷酸果糖350mg/kg,术后受体同剂量注射至处死或停跳;环孢菌素A组,受体手术当天开始肌肉注射环孢菌素A5.0mg/kg,直至处死或停跳;1,6-二磷酸果糖+环孢菌素A组,按1,6-二磷酸果糖组与环孢菌素A组联合进行给药。术后5d采用TUNEL法测定各组5例移植后心室肌细胞凋亡指数,其余5例观察移植心脏存活时间。结果:纳入Wistar大鼠(供体)和SD大鼠(受体)各40只,40只受体均进入结果分析。①1,6-二磷酸果糖组、环孢菌素A组、1,6-二磷酸果糖+环孢菌素A组大鼠移植心脏心室肌细胞凋亡指数较对照组明显降低(分别为8.06±0.70,6.63±0.52,3.12±0.45,12.07±1.32,P<0.05),1,6-二磷酸果糖+环孢菌素A组降低尤其明显(P<0.01)。②1,6-二磷酸果糖组、环孢菌素A组、1,6-二磷酸果糖+环孢菌素A组大鼠移植心脏生存时间高于对照组,差异有显著性意义[分别为(12.4±1.1),(11.6±1.1),(18.0±1.0),(6.0±1.0)d,P<0.05],1,6-二磷酸果糖+环孢菌素A组更加显著(P<0.01)。结论:1,6-二磷酸果糖是细胞能量代谢的重要中间产物,可以减少移植心肌细胞凋亡,对促进移植心脏复跳、保护心功能、延长移植心脏生存时间均有显著效果,可以作为心脏移植围手术期与环孢素A协同用药。

AIM： To observe the effect of the fructose-1 ,6-diphosphate （FDP） used solely and combined with the cyclosporine A （CsA） on apoptosis in setting up the model of cardiac allograft in rats in acute rejection phase. METHODS： The experiment was completed in the Experiment Center of Nanjing General Hospital of Nanjing Military Area Command of Chinese PLA from April to December 2004. Totally 40 Wistar rats （donator） and 40 SD rats （receptor） were selected. After hearts of Wistar rats were implanted into abdominal cavity of SD rats according to modified Ono operation, they were divided into four groups： control group, intervention before and after operation; FDP group, 350 mg/kg FDP was administrated in donators daily by vein injection at day 5 before operation; receptors were injected with the same volume till killed or arrested; CsA group, 5.0 mg/kg CsA was injected immediately by muscle injection till killed or arrested; FDP plus CsA group, the rats were administrated based on that in the FDP group and CsA group. Five days after operation apoptotic index of ventricular myocytes in 5 rats was measured with TUNEL method. Survival time of the replanted heart was observed in the other 5 rats. RESULTS： Of the 40 Wistar rats （donator） and 40 SD rats （receptor）, 40 receptors were involved in the result analysis. ①The apoptotic index in the three experimental groups was significantly lower than that in the control group （8.06±0.70,6.63±0.52,3.12±0.45,12.07±1.32,P 〈 0.05）, especially in the FDP plus CsA group （P 〈 0.01 ）. ②Survival time the three experimental groups was significantly longer than that in the control group, which had significant difference [ （ 12.4± 1.1 ）, （ 11.6± 1.1 ） , （18.0±1.0）, （6.0±1.0） days,P 〈 0.05], especially in the FDP plus CsA group （P 〈 0.01 ）. CONCLUSION： FDP is a very important mesne production in energy metabolism, which can reduce cell apoptosis and has positive effect on replanted heart, including advancing heart self-recovery, protecting function, and prolonging survival time. So it can be used as a concurrent drug with CsA during the perioperative stage of the heart transplantation.