UW液和HTK液低温保存大鼠后肢效果的对比研究

Contrastive study on cryopreservation of rat amputated limb with UW solution and HTK solution

目的比较UW液和HTK液4℃常规低温保存大鼠离断后肢的效果。方法将30只Wistar大鼠随机分为UW液组、HTK液组及对照组（每组10只），形成后肢离断模型后，分别采用UW液、HTK液及生理盐水灌注离断肢体后。4℃保存。收集不同时间点的肌肉匀浆液及组织，观察生化指标和光镜下结构变化。结果与对照组比较，uw液和HTK液组于相同时间点发生病理损伤程度轻，前者又较后者发生病理损伤程度轻。保存12h，3组均出现线粒体肿胀空泡变性，UW液组肌纤维损伤最轻。保存24h，对照组出现肌纤维断裂成碎片，损伤最为严重，UW液组和HTK液组变化基本类似，对照组、HTK液组和UW液组ALP（U／L）分别为3．62±1．32、3．37±0．84和2．68±1．59，ALT（U／L）分别为542．25±129．36、521．82±97．32和462．53±74．18，AST（U／L）分另0为2200．12±687．61、2002．20±632．27和1742．87±396．09，CK（U／L）分另0为313190．83±42041．32、283173．50±31488．87和271319．67-4-41147．52；LDH（U／L）分别为32409．50±4253．20、30382．50±2337．18和30047．83±4628．78，GLU（mmol／L）分别为0．242±0．041、0．240±0．044和0．252±0．049；LAC（mmol／L）分别为1．790±0．160、1．792±0．196和1．993±0．366，SOD（U／mL）分别为80．82±9．46、91．62±14．97和73．71±10．60．uw液组与对照组比较，ALP和GLU变化差异有统计学意义（P〈0．05）；UW液组SOD与HTK液组比较，差异有统计学意义（P〈0．05），其余生化指标间差异无统计学意义（P〉0．05）。总体生化指标UW液较HTK液结果好。结论应用器官保存液对离断肢体进行灌注保护可以有效的减缓骨骼肌组织损伤，延长组织活性：UW液较HTK液对离断肢体的保护效果更好。

Objective To compare the effect on preserving rat amputated extremities during the cold preser- vation between the UW solution and HTK solution. Methods Thirty healthy adult Wistar rats were randomly divid- ed into 3 groups （10 rats in each group） for producing the models of amputated limbs of rats. The three groups were separately irrigated the amputated limbs with UW solution, HTK solution and saline from femoral artery. After irriga- tion, the extremities were preserved in the refrigerator at 4℃ Samples of skeleton muscle tissue were taken for patho- logical and biochemical examination every 6 hours since the amputation. Results After perfusion, HE： changes of organizational structures of skeletal muscle were lighter in experimental groups than in control group （saline-perfusion group） at the same time. While samples in UW solution-perfusion group has lighter changes of organizational structure than those in HTK solution-perfusion group. TEM： at 12 h, all the three groups have shown the mitochondria edema. At 24 h, the myofibril in saline-peffusion group become fragments. Biochemical examination： at 24 h, indexes in saline- perfusion group, HTK solution group and UW solution group were： ALP （U/L） 3.62 ± 1.32, 3.37 ± 0.84 and 2.68 ± 1.59, respectively; ALT（U/L） 542.25 ± 129.36, 521.82 ± 97.32 and 462.53 ± 74.18, respectively; AST （U/L） 2200.12 ± 687.61, 2002.20 ± 632.27 and 1742.87 ± 396.09, respectively; CK（U/L） 313190.83 ± 42041.32, 283173.50 ± 31488.87 and 271319.67 ± 41147.52, respectively; LDH （U/L） 32409.50 ± 4253.20, 30382.50 ± 2337.18 and 30047.83 ± 4628.78, respectively; GLU（mmol/L） 0.242 ± 0.041,0.240 ± 0.044 and 0.252 ± 0.049, respectively; LAC （mmol/L） 1.790 ± 0.160, 1.792±0.196 and 1.993 ± 0.366, respectively; SOD（U/mL） 80.82 ± 9.46, 91.62 ± 14.97 and 73.71 ± 10.60, respectively. There were statistical difference among the UW solution-perfusion group and centrol group in ALP and GLU （P 〈 0.05）. UW solution-perfusion group has better resuhs than HTK solution-perfusion group in SOD （P 〈 0.05）. Conclusion Amputated limbs irrigated with organ preservation solution as a way to protect skeletal muscle had great effects on lightening tissue damage and keeping tissue active. Between the two usual organ preservation solution, UW solution had a stronger protective effects on the amputated limbs than HTK solution.