UW液与HTK液灌注保存大鼠离断骨骼肌效果比较

Pathological changes of amputated limbs of rats preserved with UW solution or HTK solution perfusion

目的比较UW液与HTK液,挑选出更适合保存离断肢体的灌注液体。方法选取健康成年Wistar大鼠（雌性）40只（体质量250～300 g）,随机分为4组,UW液灌注组、HTK液灌注组、0.9%氯化钠注射液灌注组、不灌注对照组,每组10只,共形成80只离断肢体模型。0.9%氯化钠注射液灌注组、HTK液灌注组、UW液灌注组均以12 ml/h速度自股动脉灌注对应灌注液1 h。所有离断肢体模型自离断后均放置在4℃的冰箱内保存24 h。在离断6 h、12 h、18 h、24 h时取肌肉组织行伊红-苏木素染色、磷钨酸苏木素染色观察骨骼肌病理变化,并用Image-pro plus软件统计HE染色细胞面积比例。结果各时间点,3个灌注组较不灌注组骨骼肌组织病理变化轻微,其中UW液组病理变化最轻。细胞面积比例各组间均有统计学差异（P〈0.05）,各时间点不灌注组的细胞面积比例为43.93%±12.16%（6 h）、38.41%±6.54%（12h）、35.27%±7.75%（18 h）、36.24%±7.09%（24 h）,均为最低,6～12 h 0.9%氯化钠注射液组细胞面积比例最大（6 h为92.54%±3.26%,12 h为59.35%±4.64%）,18～24 h UW液组细胞面积比例最大（18 h为51.95%±10.35%,24 h为47.56%±7.04%）。结论对大鼠离断肢体进行一次性灌注,24 h内UW液的灌注保存效果最好。

Objective To find out a more suitable solution for amputated limbs preservation by comparing two common organ preservation solution. Methods Forty adult female healthy Wistar rats, weighted 250- 300 g, were provided by the Experimental Animal Center of the Academy of Science and Military of PLA. Rats were randomly divided into 4 groups（10 rats in each group） to establish the models of amputated limbs. Rats in the first group were perfused with saline, the second group with HTK solution, the third group with UW solution, and rats in the fourth group were not perfused with any solution, as the control group. All groups were preserved in the refrigerator at 4℃ for 24 h, and the first three groups were perfused at the speed of 12 ml/h for 1 h just after the amputation. Specimens of muscle tissue were taken for pathological（HE and PTAH） examination every 6 hours. Results During the cryopreservation, the pathological changes of muscle tissue were less severe in perfusion groups than non-perfusion group. Among the three perfusion groups, the third group（with UW solution） had the lowest degree of pathological change. The area of cell had statistically significant differences between each group, and non-perfusion group had the smallest area of cell [43.93%±12.16%（6 h）,38.41%±6.54%（12 h）, 35.27%±7.75%（18 h）, 36.24%±7.09%（24 h）, P 0.05]. The saline-perfusion group had the largest area of cell within 12 hours [92.54%±3.26%（6 h）, 59.35%±4.64%（12 h）], while after 12 h, UW-perfusion group reached to the top [51.95%±10.35%（18 h）, 47.56%±7.04%（24 h）]. Conclusion Compared with HTK solution and saline, UW solution has the best protective effect on amputated limbs during 24 h preservation.