铁离子螯合剂对肝细胞冷缺血再灌注损伤的保护机制研究

Protective effect of iron chelator on hepatocyte underwent cold ischemia/reperfusion injury

目的研究铁螯合剂去铁胺(DFO)对模拟冷缺血再灌注后肝细胞内铁离子浓度及细胞凋亡的影响。方法培养肝细胞株HL-7702,随机分为对照组和DFO干预组。对照组未予DFO处理,DFO干预组按100、50和25μmol/L分为高、中、低3个浓度组,每组各8个样本,建立模拟冷缺血再灌注模型,分别检测各组细胞内Fe3+离子浓度,MDA含量,细胞凋亡水平,以及各组上清液乳酸脱氢酶(LDH)水平。结果肝细胞经冷缺血8 h,DFO干预高、中、低浓度组Fe3+浓度分别为(1.36±0.14)、(1.92±0.24)和(2.09±0.28)mmol/L,均显著低于对照组(2.58±0.27)mmol/L,(P<0.05);DFO高、中、低浓度组肝细胞凋亡率分别为(6.37±1.28)%、(14.07±2.61)%和(16.07±1.71)%,均低于对照组(20.53±1.58)%,(均P<0.01)。DFO高、中、低浓度组肝细胞内MDA含量分别为(1.83±0.63)、(2.60±0.26)和(2.74±0.32)nmol/m L,均显著低于对照组(3.57±0.45)nmol/m L(P<0.01)。而再灌注8 h时,DFO高、中、低浓度组肝细胞内MDA含量分别为(2.48±0.36)、(3.07±0.35)和(3.56±0.25)nmol/m L,均显著低于对照组(4.16±0.24)nmol/m L(P<0.05);在LDH水平测定中,DFO高、中、低浓度组上清液LDH水平分别为(216.67±22.22)、(263.68±7.16)和(297.57±21.06)u/L,均低于对照组(354.51±31.02)u/L(P<0.05)。结论铁螯合剂去铁胺可以通过降低冷保存时Fe3+浓度,从而减少肝细胞凋亡发生以达到减轻肝细胞冷缺血损害的目的。

[ Objective ] To investigate the effect of iron chelator-Desferrioxamine （DFO） on intracellular iron ion concentration （Fe^3＋） and apoptosis of hepatocyte. [ Methods] HL-7702 Hepatocytes were divided into control group and DFO group. In the DFO group, three subgroups were divided according to DFO level （n =8）. The level of intra- cellular Fe3＋ level, MDA level, apoptosis index as well as the LDH level in supernatant were determined using mim- icking hepatoeytes cold ischemia/warm repferfusion model. [Result] After 8h cold isehemia, Fe3 ＋ level in DFO high-level （HL）, middle-level （ML） and low-level （LL） subgroups were （1.36±0.14）, （1.92±0.24）, （2.09±0.28） mmol/ L, all of which lowered than that in control group statistically （2.58±0.27） mmol/L, （P 〈0.01）. The apoptosis ratio in HL, ML, LL subgroups were （6.37±1.28）, （14.07±2.61）, （16.07±1.71）%. They also all were lowered that in control group statistically（20.53±1.58%）, （P 〈0.01）. The MDA level in HL, ML, LL subgroups were （1.83±0.63）, （2.60±0.26）,（2.74±0.32） nmol/mL, all of which lowered than that in control group statistically （3.57±0.45） nmol/mL （P 〈0.01）. After 8 h reperfusion, The MDA level in HL, ML, LL subgroups were （2.48±0.36）, （3.07±0.35）, （3.56±0.25） nmol/mL which were statistically lowered compared with control group （4.16±0.24） nmol/mL （P 〈0.05）. The LDH test showed the level of LDH in HL, ML, LL subgroups were （216.67±22.22）, （263.68±7.16）, （297.57±21.06） U/L respectively, which were statistically lowered compared with control group （354.51±31.02） U/L （P 〈0.05）. [ Conclusion ] The iron chelator Desferrioxamine could attenuate the cold ischemia injury by reducing the Fe3＋ level so that control of the apoptosis.