一氧化氮在犬离体胰腺低温灌注与保存中的作用

Protective effects of nitric oxide on the reperfusion injury of isolated pancreas with hypothermic perfusion and preservation in canines

目的探讨一氧化氮(NO)对犬胰腺低温灌注及保存中缺血再灌注损伤的影响。方法在EC(EuroCollin)液中分别加用L精氨酸(LArg,200mg/kg体重)、N硝基L精氨酸甲酯(LNAME,10mg/kg体重)和生理盐水,分别对犬离体胰腺节段进行低温灌注保存(灌注量为30～50ml,温度为0～4℃,时间为24h),应用犬胰节段移植模型,测定移植后犬血清中脂肪酶、淀粉酶含量,测定移植胰腺组织中髓过氧化酶(MPO)活性、一氧化氮(NO)含量、一氧化氮合酶(NOS)及诱导型一氧化氮合酶(iNOS)mRNA的表达情况。并进行组织学观察。结果血清脂肪酶,LArg处理组<对照组<LNAME处理组,差异有统计学意义(P<0.05)。移植后12h,胰腺组织中NO含量:LArg处理组(0.51±0.11)μmol/L>对照组(0.31±0.06)μmol/L>LNAME处理组(0.17±0.04)μmol/L;NOS活性:LArg处理组(3.13±0.19)U/ml>对照组(2.37±0.20)U/ml>LNAME处理组(1.68±0.20)U/ml;iNOSmRNA的表达:LArg处理组(0.89±0.22)μmol/L>对照组(0.63±0.18)μmol/L>LNAME处理组(0.49±0.20)μmol/L,差异有统计学意义(P<0.05);LArg处理组胰小叶间质水肿和胰小叶内中性粒细胞浸润较轻。结论NO对犬胰腺低温灌注及保存中缺血再灌注损伤具有保护作用。

Objective To study the protective effects of nitric oxide （NO） on reperfusion injury following segmental pancreatic transplantation in canines. Methods Segmental pancreatic transplantation was carried out. In the experimental groups, the L-Arg （200 mg/kg） or L-NAME（10 mg/kg） as an additive agent was added into the traditional EC solution （30-50 ml）. The isolated pancreas of dogs was flushed and stored at 0-4℃ for 24 h. In the control group, the EC solution was used. Serum concentrations of amylase and lipase were determined. In the pancreas graft, the contents of NO, NOS, iNOS mRNA and MPO were examined. Histological observation was performed. Results The content of NO in the L-Arg group （0.51 ± 0.11）μmol/L was higher than in the control group （0.31 ± 0.06） /μmol/L, while （0.17 ± 0.04） μmol/L in the L-NAME group was lower than in the control group （P〈 0.05）. The activitiy of NOS in L-Arg group （3.13 ± 0.19） U/ml was higher than in the control group （2.37 ± 0.20） U/ml, while in the L-NAME group, that （1.68± 0.20） U/ml was lower than in control group （P〈0.05） .The expression of iNOS mRNA in L-Arg group （0.89 ± 0.22） μmol/L was higher than in the control group （0.63 ± 0.18）μmol/L, while in the L-NAME group, that （0.49 ± 0.20） μmoL/L was lower than in control group （P〈 0.05）. The pancrease inflammation was ameliorated when L-Arg was administered, while the pancrease damage was aggravated when L-NAME was administered. Conclusion NO has protective effects on isolated pancreas with hypothermic perfusion and preservation.