依达拉奉对大鼠再植断肢的保护作用
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摘要
目的：本实验通过建立大鼠断肢再植模型,检测依达拉奉对血清中超氧化物歧化酶(SOD)、丙二醛(MDA)及骨骼肌线粒体Na+-K+-ATP酶、Ca2+-ATP酶活性的影响及光镜、电镜下骨骼肌结构的改变,解明依达拉奉对大鼠再植后断肢的保护作用。
方法：清洁级健康雌性SD大鼠40只,随机分为4组,每组10只：S组假手术组,M组断肢再植模型组,E1组依达拉奉低剂量组,E2组依达拉奉高剂量组。M组：大鼠麻醉成功后固定,右后肢常规皮肤消毒后沿股血管走行方向纵行切开皮肤,显露并游离股动静脉及股神经,钝性分离各层组织,于右股骨中段水平切断所有股部肌肉至股骨,只保留血管。右后肢驱血后以血管夹夹闭右侧股动静脉,室温下放置3h,放松血管夹,形成再灌注,缝合肌肉、神经及皮肤。术野用生理盐水浸湿的无菌纱布覆盖。再灌注前各组均由尾静脉给予肝素钠1000U/kg。E1、E2组除与M组上述操作相同外,于再灌注前分别由尾静脉注射依达拉奉0.5mg/kg和1mg/kg。S组和M组给予等量的生理盐水。S组仅游离出股动静脉,旷置3h后缝合皮肤。
每组于再灌注5h和24h分别经颈外静脉取血各lml,静置后离心,测定血清超氧化物歧化酶(SOD)的活性及丙二醛(MDA)的含量。并于再灌注24h取右后肢腓肠肌,提取线粒体测定骨骼肌线粒体Na+-K+-ATP酶、Ca2+-ATP酶活性的变化。另取右后肢胫前肌在光镜和电镜下观察骨骼肌的结构变化。
结果：40只大鼠全部进入结果分析。
1大体形态观察：S组右后肢肌肉、皮肤红润温暖。M组右后肢肌肉、皮肤颜色稍暗红,温度稍低。E1组右后肢外观改变较M组减轻。E2组右后肢外观较M组有明显的改善。
2骨骼肌组织切片观察：S组肌纤维基本正常,间质无水肿,未见炎细胞浸润。M组肌纤维结构紊乱,崩解,血管扩张充血,间质水肿,有较多的炎性细胞浸润。E1组肌纤维轻度水肿,间质稍水肿,有炎细胞浸润。E2组间质稍水肿,肌间散在炎细胞浸润。
3透射电镜观察：S组肌原纤维排列整齐,Z线清晰。线粒体嵴致密,膜完整,无肿胀。M组肌原纤维部分断裂,Z线较模糊,线粒体明显肿胀,嵴紊乱,部分空泡化。E,组肌原纤维排列较整齐,Z线稍模糊,含有较多的脂滴,线粒体部分空泡化。E2组肌原纤维排列较整齐,Z线较清晰,含有少量脂滴,线粒体部分空泡化。
4血清SOD活性及MDA含量：
血清SOD活性：与S组比,M、E1、E2组SOD活性均降低(P＜0.05)。与M组比,E1、E2组SOD活性均升高(P<0.05)。与E1组比,E2组SOD活性升高(P＜0.05)。再灌注24h时,M、E1、E2组SOD活性较5h时低。
血清MDA含量：与S组比,M、E1、E2组MDA含量均升高(P＜0.05)。与M组比,E1、E2组MDA含量均降低(P＜11.05)。与E1组比,E2组MDA含量降低(P＜0.05)。再灌注24h时,M、E1、E2组MDA含量较5h时高。
S组再灌注24h时上述两指标与再灌注5h时比较差异无统计学意义(P＞0.05)。
5骨骼肌线粒体Na+-K+-ATP酶、Ca2+-ATP酶活性：与S组比,M、E1、E2组Na+-K+-ATP酶、Ca2+-ATP酶均降低(P＜0.05)。与M组比,E1、E2组Na+-K+-ATP酶、Ca2+-ATP酶均升高(P＜0.05)。与E1组比,E2组Na+-K+-ATP酶. Ca2+-ATP酶升高(P＜0.05)。
结论：
1依达拉奉可减轻大鼠肢体缺血再灌注的损伤,对再植后的断肢具有保护作用,且高剂量组优于低剂量组。
2其机制可能是增高了SOD活性,降低MDA含量,提高骨骼肌线粒体内Na+.K+-ATP酶、Ca2+-ATP酶活性。
Objective:In order to study the protective effects of Edaravone on the replanted limb of rats, a model of limb replantation was established. By detecting the effects of Edaravone to serum Superoxidedismutase (SOD), Malondialdehyde (MDA), activity of Na+-K+-ATPase and Ca2+-ATPase of mitochondrion in skeletal muscle, and the structural changes of skeletal muscle was observed with optical microscope and electron microscope.
Methods:Forty female Sprague-Dawley (SD) rats were randomly divided into four groups of ten rats each. Sham group (S), model group (M), low concentration of edaravone group (E1), high concentration of edaravone group (E2). M group was the model of limb replantation of rats. The rats were fixed following anesthetized. After the right posterior limb was disinfected, along the longitudinal direction on femoral vessels cut skin, the femoral artery, vein and nerve were exposed and dissociated. Blunt separation layers of organizations, in middle level of right femur cut off all muscles, only preserve vessels. After the blood was drove, the femoral artery and femoral vein were occluded by vascular clamp. The right posterior limb was put in the room temperature for 3h, then the vascular clamp was removed for reperfusion, suture the muscle, nerve and skin. The operative field was covered with saline soaked sterile gauze. Each group was given heparin sodium 1000U/kg to heparinization via the caudal vein before reperfusion. The rats in group E1 and E2 not only received the operation mentioned above, but also injected edaravone respectively with the concentration of 0.5mg/kg and lmg/kg via the caudal vein before reperfusion. The same volume of saline was injected in group S and M. In group S, the right posterior limb only dissociates the femoral artery and vein, put in the room temperature for 3h, then suture the skin. At the time 5h and 24h reperfusion,1 ml blood of each rat was taken via the external jugular vein. Serum Superoxidedismutase (SOD) and Malondialdehyde (MDA) were determined. At 24h take the right posterior limb gastrocnemius, extract mitochondrial to determine the changes of Na+-K+-ATPase and Ca2+-ATPase. Take the right posterior limb tibial muscle to detect the structural changes of skeletal muscle with optical microscope and electron microscope.
Results:40 healthy SD rats included in the experiment entered the final analysis.
1 The general morphological observation:In group S, right posterior limb muscle and skin ruddy and warm. In the group M, the color of right posterior limb muscle and skin was slightly dark red, and the temperature was slightly lower. In group E1, the appearance of the right posterior limb was alleviative to group M. The appearance in group E2 was more alleviative than group M.
2 For the histological section:In group S, the muscular fiber was normal, there was no edema in the interstitial, and there was no inflammatory cells infiltration. In group M, the muscular fiber was chaotic and ruptured. The blood vessel was dilated and congestive. The interstitial was edema, and there was infiltration with many inflammatory cells. In group E1, the muscular fiber and interstitial were slightly edema. There was inflammatory cell infiltration. In group E2, the interstitial was slightly edema, and there were scattered inflammatory cells infiltrate.
3 Observations from transmission electron microscope:The arrangement of myofibril in group S was relatively regular. Z line was clear, mitochondrial crista was compact, membrane was intact and without swelling. Parts of myofibril in group M were broken. Z line was unclear or disappeared, mitochondria were highly swollen and mitochondrial crista was in chaos, and parts had been vacuoles. In group E1, the arrangement of myofibril was relatively regular. Z line was a little obscured. There have major lipid droplets. Parts of the mitochondria were vacuolated. In group E2, the arrangement of myofibril was relatively regular. Z line was clear. There have small amounts of lipid droplets. Parts of the mitochondria were vacuolated.
4 The activity of SOD and the content of MDA in serum:
Serum SOD activity:Activity of SOD in group M, E1 and E2 was lower than group S (p＜0.05). The activity of SOD in group E1 and E2 was higher than group M (p＜0.05). The activity of SOD in group E2 was higher than group E1 (p＜0.05). The activity of SOD at 24h of reperfusion in group M, E1 and E2 was lower than 5h.
Serum MDA content:Content of MDA in group M, E1 and E2 was higher than group S (p＜0.05). The content of MDA in group E1 and E2 was lower than group M (p＜0.05). The content of MDA in group E2 was lower than group E1 (p＜0.05). The content of MDA at 24h of reperfusion in group M, E1 and E2 was higher than 5h.
There was no significant difference of SOD and MDA at 24h of reperfusion compared with that 5h in group S (p>0.05).
5 Activity of mitochondria Na+-K+-ATPase and Ca2+-ATPase:The activity of Na+-K+-ATPase and Ca2+-ATPase in group M, E1 and E2 was lower than group S (p＜0.05). The activity of Na+-K+-ATPase and Ca2+-ATPase in group E1 and E2 was higher than group M (p<0.05). The activity of Na+-K+-ATPase and Ca2+-ATPase in group E2 was higher than group E1 (p＜0.05).
Conclusion:
1 Edaravone can reduce ischemia reperfusion injury of rats, have the protective effect on replanted limb.
2 The mechanism may be increased in the activity of SOD, mitochondria Na+-K+-ATPase and Ca2+-ATPase, decreased the content of MDA.
方法：清洁级健康雌性SD大鼠40只,随机分为4组,每组10只：S组假手术组,M组断肢再植模型组,E1组依达拉奉低剂量组,E2组依达拉奉高剂量组。M组：大鼠麻醉成功后固定,右后肢常规皮肤消毒后沿股血管走行方向纵行切开皮肤,显露并游离股动静脉及股神经,钝性分离各层组织,于右股骨中段水平切断所有股部肌肉至股骨,只保留血管。右后肢驱血后以血管夹夹闭右侧股动静脉,室温下放置3h,放松血管夹,形成再灌注,缝合肌肉、神经及皮肤。术野用生理盐水浸湿的无菌纱布覆盖。再灌注前各组均由尾静脉给予肝素钠1000U/kg。E1、E2组除与M组上述操作相同外,于再灌注前分别由尾静脉注射依达拉奉0.5mg/kg和1mg/kg。S组和M组给予等量的生理盐水。S组仅游离出股动静脉,旷置3h后缝合皮肤。
每组于再灌注5h和24h分别经颈外静脉取血各lml,静置后离心,测定血清超氧化物歧化酶(SOD)的活性及丙二醛(MDA)的含量。并于再灌注24h取右后肢腓肠肌,提取线粒体测定骨骼肌线粒体Na+-K+-ATP酶、Ca2+-ATP酶活性的变化。另取右后肢胫前肌在光镜和电镜下观察骨骼肌的结构变化。
结果：40只大鼠全部进入结果分析。
1大体形态观察：S组右后肢肌肉、皮肤红润温暖。M组右后肢肌肉、皮肤颜色稍暗红,温度稍低。E1组右后肢外观改变较M组减轻。E2组右后肢外观较M组有明显的改善。
2骨骼肌组织切片观察：S组肌纤维基本正常,间质无水肿,未见炎细胞浸润。M组肌纤维结构紊乱,崩解,血管扩张充血,间质水肿,有较多的炎性细胞浸润。E1组肌纤维轻度水肿,间质稍水肿,有炎细胞浸润。E2组间质稍水肿,肌间散在炎细胞浸润。
3透射电镜观察：S组肌原纤维排列整齐,Z线清晰。线粒体嵴致密,膜完整,无肿胀。M组肌原纤维部分断裂,Z线较模糊,线粒体明显肿胀,嵴紊乱,部分空泡化。E,组肌原纤维排列较整齐,Z线稍模糊,含有较多的脂滴,线粒体部分空泡化。E2组肌原纤维排列较整齐,Z线较清晰,含有少量脂滴,线粒体部分空泡化。
4血清SOD活性及MDA含量：
血清SOD活性：与S组比,M、E1、E2组SOD活性均降低(P＜0.05)。与M组比,E1、E2组SOD活性均升高(P<0.05)。与E1组比,E2组SOD活性升高(P＜0.05)。再灌注24h时,M、E1、E2组SOD活性较5h时低。
血清MDA含量：与S组比,M、E1、E2组MDA含量均升高(P＜0.05)。与M组比,E1、E2组MDA含量均降低(P＜11.05)。与E1组比,E2组MDA含量降低(P＜0.05)。再灌注24h时,M、E1、E2组MDA含量较5h时高。
S组再灌注24h时上述两指标与再灌注5h时比较差异无统计学意义(P＞0.05)。
5骨骼肌线粒体Na+-K+-ATP酶、Ca2+-ATP酶活性：与S组比,M、E1、E2组Na+-K+-ATP酶、Ca2+-ATP酶均降低(P＜0.05)。与M组比,E1、E2组Na+-K+-ATP酶、Ca2+-ATP酶均升高(P＜0.05)。与E1组比,E2组Na+-K+-ATP酶. Ca2+-ATP酶升高(P＜0.05)。
结论：
1依达拉奉可减轻大鼠肢体缺血再灌注的损伤,对再植后的断肢具有保护作用,且高剂量组优于低剂量组。
2其机制可能是增高了SOD活性,降低MDA含量,提高骨骼肌线粒体内Na+.K+-ATP酶、Ca2+-ATP酶活性。
Objective:In order to study the protective effects of Edaravone on the replanted limb of rats, a model of limb replantation was established. By detecting the effects of Edaravone to serum Superoxidedismutase (SOD), Malondialdehyde (MDA), activity of Na+-K+-ATPase and Ca2+-ATPase of mitochondrion in skeletal muscle, and the structural changes of skeletal muscle was observed with optical microscope and electron microscope.
Methods:Forty female Sprague-Dawley (SD) rats were randomly divided into four groups of ten rats each. Sham group (S), model group (M), low concentration of edaravone group (E1), high concentration of edaravone group (E2). M group was the model of limb replantation of rats. The rats were fixed following anesthetized. After the right posterior limb was disinfected, along the longitudinal direction on femoral vessels cut skin, the femoral artery, vein and nerve were exposed and dissociated. Blunt separation layers of organizations, in middle level of right femur cut off all muscles, only preserve vessels. After the blood was drove, the femoral artery and femoral vein were occluded by vascular clamp. The right posterior limb was put in the room temperature for 3h, then the vascular clamp was removed for reperfusion, suture the muscle, nerve and skin. The operative field was covered with saline soaked sterile gauze. Each group was given heparin sodium 1000U/kg to heparinization via the caudal vein before reperfusion. The rats in group E1 and E2 not only received the operation mentioned above, but also injected edaravone respectively with the concentration of 0.5mg/kg and lmg/kg via the caudal vein before reperfusion. The same volume of saline was injected in group S and M. In group S, the right posterior limb only dissociates the femoral artery and vein, put in the room temperature for 3h, then suture the skin. At the time 5h and 24h reperfusion,1 ml blood of each rat was taken via the external jugular vein. Serum Superoxidedismutase (SOD) and Malondialdehyde (MDA) were determined. At 24h take the right posterior limb gastrocnemius, extract mitochondrial to determine the changes of Na+-K+-ATPase and Ca2+-ATPase. Take the right posterior limb tibial muscle to detect the structural changes of skeletal muscle with optical microscope and electron microscope.
Results:40 healthy SD rats included in the experiment entered the final analysis.
1 The general morphological observation:In group S, right posterior limb muscle and skin ruddy and warm. In the group M, the color of right posterior limb muscle and skin was slightly dark red, and the temperature was slightly lower. In group E1, the appearance of the right posterior limb was alleviative to group M. The appearance in group E2 was more alleviative than group M.
2 For the histological section:In group S, the muscular fiber was normal, there was no edema in the interstitial, and there was no inflammatory cells infiltration. In group M, the muscular fiber was chaotic and ruptured. The blood vessel was dilated and congestive. The interstitial was edema, and there was infiltration with many inflammatory cells. In group E1, the muscular fiber and interstitial were slightly edema. There was inflammatory cell infiltration. In group E2, the interstitial was slightly edema, and there were scattered inflammatory cells infiltrate.
3 Observations from transmission electron microscope:The arrangement of myofibril in group S was relatively regular. Z line was clear, mitochondrial crista was compact, membrane was intact and without swelling. Parts of myofibril in group M were broken. Z line was unclear or disappeared, mitochondria were highly swollen and mitochondrial crista was in chaos, and parts had been vacuoles. In group E1, the arrangement of myofibril was relatively regular. Z line was a little obscured. There have major lipid droplets. Parts of the mitochondria were vacuolated. In group E2, the arrangement of myofibril was relatively regular. Z line was clear. There have small amounts of lipid droplets. Parts of the mitochondria were vacuolated.
4 The activity of SOD and the content of MDA in serum:
Serum SOD activity:Activity of SOD in group M, E1 and E2 was lower than group S (p＜0.05). The activity of SOD in group E1 and E2 was higher than group M (p＜0.05). The activity of SOD in group E2 was higher than group E1 (p＜0.05). The activity of SOD at 24h of reperfusion in group M, E1 and E2 was lower than 5h.
Serum MDA content:Content of MDA in group M, E1 and E2 was higher than group S (p＜0.05). The content of MDA in group E1 and E2 was lower than group M (p＜0.05). The content of MDA in group E2 was lower than group E1 (p＜0.05). The content of MDA at 24h of reperfusion in group M, E1 and E2 was higher than 5h.
There was no significant difference of SOD and MDA at 24h of reperfusion compared with that 5h in group S (p>0.05).
5 Activity of mitochondria Na+-K+-ATPase and Ca2+-ATPase:The activity of Na+-K+-ATPase and Ca2+-ATPase in group M, E1 and E2 was lower than group S (p＜0.05). The activity of Na+-K+-ATPase and Ca2+-ATPase in group E1 and E2 was higher than group M (p<0.05). The activity of Na+-K+-ATPase and Ca2+-ATPase in group E2 was higher than group E1 (p＜0.05).
Conclusion:
1 Edaravone can reduce ischemia reperfusion injury of rats, have the protective effect on replanted limb.
2 The mechanism may be increased in the activity of SOD, mitochondria Na+-K+-ATPase and Ca2+-ATPase, decreased the content of MDA.
引文
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11 Lepore DA. Nitric oxide synthase-independent generation of nitric oxide in muscle ischemia--reperfusion injury. Nitric Oxide 2000,4(6):541-545
12 Roth E. The impact of L-arginine-nitric oxide metabolism on ischemia/reperfusion injury. Curr Opin Clin Nutr Metab Care 1998,1(1):97-99
13 Hatoko M, Tanaka A, Kuwahara M, et al. Difference of molecular response to ischemia-reperfusion of rat skelletal muscle as a function of ischemic time:study of the expression of p53,p21(WAF-1), Bax protein, and apoptosis. Ann Plast Surg 2002,48(1):68-74
14杨军,胡新华,张强,等.骨骼肌缺血再灌注后肺细胞间黏附分子1的表达[J].中国医科大学学报,2004,33(4)：296-298
15 Fitzal F, Delano FA, Young C, et al. Early capillary no-reflow during low-flow reperfusion after hind limb ischemia in the rat. Ann Plast Surg 2002,49(2):170-180
16张润峰,李霞.丹参改善缺血再灌注血液流变学异常的作用机制研究[J].华西医学,2004,19(2)：341-342
17孙向东,况炜,陈莹莹,等.过氧化氢的血管效应及其作用机制[J].现代实用医学,2005,17(2)：124-127
18 Bonheur JA, Albadawi H, Patton GM, et al. A noninvasive murine model of hind limb ischemia-reperfusion injury. J Surg Res 2004,116(1):55-63
19符庆瑛,李著华.肾缺血再灌注损伤中微血管损伤防治的研究进展[J].微循环学杂志,2004,14(1)：50-52
20 Beyerdorf F, et, al Thorac Cardiovasc Surg,1991,39(2):19
21 Tamai S, et al Phst Reconstr Surg 1990,46(3):219
22 Shenton OW, et al Jhone joint J,1990,72A:1405
23 Haimocivi H, I Camiovasc Surg,1990,31 (3);318
24 Gregory MA, et, al S Afr Med J,1991,79(16):307
25 Barie PS, et, al J Surg Res,1988,44:284
26 Okad k. Microvasc Res,1990,39(2):156
27张新宇,王力平,樊东升,等.重组组织型纤溶酶原激活剂与依达拉奉联合应用治疗超早期脑梗死.中华神经科杂志,2008,41：201-203
28 Asai T, Ohno Y, Minatoguchi S, et al. The specific free radical scavenger edaravone suppresses bleomycin-induced acute pulmonary injury in rabbits. Clin Exp Pharmacol Physiol,2007,34(1-2):22-26
29 Kawasaki T, Ishihara K, Ago Y, et al. Protective effect of the radical scavenger edaravone against methamphetamine-induced dopaminergic neurotoxicity in mouse striatum. Eur J Pharmacol,2006,542(1-3):92-99
30 Dohi K, Satoh K, Mihara Y, et al. Alkoxyl radical-scavenging activity of edaravone in patients with traumatic brain injury. J Neurotrauma, 2006,23(11):1591-1599
31 Katare GR, Shiro S. Antioxidant MCI-186 inhibits mitochondrial permeability transition pore and up regulates BCL-2 expression[J]. AJP-Heart,2003,285:2171
32段国贤,赵春秀,王艳蕾,等.参麦注射液对兔肢体缺血再灌注致远隔器官脂质过氧化损伤的保护作用.陕西医学杂志,2004,33(7)：598
33梁贵友,牛义民.内源性一氧化氮抗兔肺缺血再灌注损伤作用及其机制探讨[J].四川大学学报(医学版),2005,36(2)：246-248
34 Ito K, Ozasa H, Horikawa S. Edaravone protects against lung injury induced by intestiral ischemia/reperfusion in rat[J]. Free Radic Biol Med, 2005,38(3):369-374
35 Reyes YA, Shimoyama T. MCT-186(edaravone), a free radical scavenger, attennates ischemia-reperfusion injury and activation of phospholipase A(2) in an isolanted rat lung modEl after 18h of cold preserration[J]. Eur J Cardiothorac Surg,2006,29(3),304-311
36赵利军,董淑云.氧化应激在大鼠肢体缺血/再灌注心肌损伤中的作用[J].解放军医学杂志,2006,31(8)：802-803
37 Minhaz U,Tanaka M. Effect of MCI-186 on postischemic reperfusion injury in isolated rat[J]. Free Radic Res,1996,24(5):361
38周洪霞,张宇新,张连元,等.大鼠肢体缺血再灌注可致脑组织细胞凋亡[J]．解剖学杂志，2005,28(6):660-662
39周洪霞,张连元.大鼠肢体缺血再灌注脑组织Bax的表达变化及谷氨酸受体拮抗剂对其影响[J].华北煤炭医学院学报,2005,(7)2：142-143
40 Amemiya S, Kamiya T, Nito C, et al. Anti-apoptotic and neuroprotective effects of edaravone fol louing to unsient focalischemia in rats[J]. Eur J Pharmacol,2005,516(2):125-130
41 Yamamo T, Yuki S, Watonabe T, et al. DElayed neuronal death prevented by inhibition of increased hydroxyl radical formation in a transient cerebral ischemia[J]. Brain Res,1997,762(1-2):240-242
42王银环,董淑云.牛磺酸对大鼠肢体缺血再灌注后肝脏损伤的保护效应[J].第四军医大学学报,2006,27(22)：2047-2050
43 Tsvji K, Kwon AH, Yoshida H, et al. Free radical Scavenger (edaravone) prevents endotoxin-induced liver injury after partial hepatectomy in rats[J]. J Hepatol,2005,42(1):94-101
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1 McCord JM. The superoxide free radical:its biochemistry and pathophysiology. Surgery 1983,94(3):412-414
2 Koksal C, Bozkurt AK, CangEl U, et al. Attenuation of ischemia/reperfusion injury by N-acetylcysteine in a rat hind limb model. J Surg Res 2003,111(2):236-239
3 Erdogan D, Omeroglu S, Sarban S, et al. Prevention of oxidative stress due to tourniquet application. Analysis of the effects of local hypothermia and systemic allopurinol administration. Acta Orthop Belg 1999,65(2):164-169
4 Schlag MG, Harris KA, Potter RF. Role of leukocyte accumulation and oxygen radicals in ischemia-reperfusion-induced injury in skeletal muscle. Am J Physiol Heart Circ Physiol 2001,280(4):H1716-H1721
5 Pattwell D, McArdle A, Griffiths RD, et al. Measurement of free radical production by in vivo microdialysis during ischemia/reperfusion injury to skeletal muscle. Free Radic Biol Med 2001,30(9):979-985
6 Van Wagoner DR, Bond M. Reperfusion arrhythmias:new insights into the role of the Na(+)/Ca(2+) exchanger. J Mol Cell Cardiol 2001,33(12):2071-2074
7 Tricarico D, Capriulo R, Camerino DC. Involvement of K(Ca2+) channels in the local abnormalities and hyperkalemia following the ischemia-reperfusion injury of rat skeletal muscle. Neuromuscul Disord 2002,12(3):258-265
8 Takahashi K, Takahashi T, Suzuki T, et al. Protective effects of SEA0400, a novel and selective inhibitor of the Na+/Ca2+exchanger, on myocardial ischemia-reperfusion injuries. Eur J Pharmacol 2003,458(1-2):155-162
9 Rezende-Neto JB, Moore EE, Masuno T, et al. The abdominal compartment syndrome as a second insult during systemic neutrophil priming provokes multiple organ injury. Shock 2003,20(4):303-308
10 Harkin DW, Barros D'sa AA, McCallion K, et al. Circulating neutrophil priming and systemic inflammation in limb ischemia-reperfusion injury. Int Angiol 2001,20(1):78-89
11 Lepore DA. Nitric oxide synthase-independent generation of nitric oxide in muscle ischemia--reperfusion injury. Nitric Oxide 2000,4(6):541-545
12 Roth E. The impact of L-arginine-nitric oxide metabolism on ischemia/reperfusion injury. Curr Opin Clin Nutr Metab Care 1998,1(1):97-99
13 Hatoko M, Tanaka A, Kuwahara M, et al. Difference of molecular response to ischemia-reperfusion of rat skelletal muscle as a function of ischemic time:study of the expression of p53,p21(WAF-1), Bax protein, and apoptosis. Ann Plast Surg 2002,48(1):68-74
14杨军,胡新华,张强,等.骨骼肌缺血再灌注后肺细胞间黏附分子1的表达[J].中国医科大学学报,2004,33(4)：296-298
15 Fitzal F, Delano FA, Young C, et al. Early capillary no-reflow during low-flow reperfusion after hind limb ischemia in the rat. Ann Plast Surg 2002,49(2):170-180
16张润峰,李霞.丹参改善缺血再灌注血液流变学异常的作用机制研究[J].华西医学,2004,19(2)：341-342
17孙向东,况炜,陈莹莹,等.过氧化氢的血管效应及其作用机制[J].现代实用医学,2005,17(2)：124-127
18 Bonheur JA, Albadawi H, Patton GM, et al. A noninvasive murine model of hind limb ischemia-reperfusion injury. J Surg Res 2004,116(1):55-63
19符庆瑛,李著华.肾缺血再灌注损伤中微血管损伤防治的研究进展[J].微循环学杂志,2004,14(1)：50-52
20 Beyerdorf F, et, al Thorac Cardiovasc Surg,1991,39(2):19
21 Tamai S, et al Phst Reconstr Surg 1990,46(3):219
22 Shenton OW, et al Jhone joint J,1990,72A:1405
23 Haimocivi H, I Camiovasc Surg,1990,31 (3);318
24 Gregory MA, et, al S Afr Med J,1991,79(16):307
25 Barie PS, et, al J Surg Res,1988,44:284
26 Okad k. Microvasc Res,1990,39(2):156
27张新宇,王力平,樊东升,等.重组组织型纤溶酶原激活剂与依达拉奉联合应用治疗超早期脑梗死.中华神经科杂志,2008,41：201-203
28 Asai T, Ohno Y, Minatoguchi S, et al. The specific free radical scavenger edaravone suppresses bleomycin-induced acute pulmonary injury in rabbits. Clin Exp Pharmacol Physiol,2007,34(1-2):22-26
29 Kawasaki T, Ishihara K, Ago Y, et al. Protective effect of the radical scavenger edaravone against methamphetamine-induced dopaminergic neurotoxicity in mouse striatum. Eur J Pharmacol,2006,542(1-3):92-99
30 Dohi K, Satoh K, Mihara Y, et al. Alkoxyl radical-scavenging activity of edaravone in patients with traumatic brain injury. J Neurotrauma, 2006,23(11):1591-1599
31 Katare GR, Shiro S. Antioxidant MCI-186 inhibits mitochondrial permeability transition pore and up regulates BCL-2 expression[J]. AJP-Heart,2003,285:2171
32段国贤,赵春秀,王艳蕾,等.参麦注射液对兔肢体缺血再灌注致远隔器官脂质过氧化损伤的保护作用.陕西医学杂志,2004,33(7)：598
33梁贵友,牛义民.内源性一氧化氮抗兔肺缺血再灌注损伤作用及其机制探讨[J].四川大学学报(医学版),2005,36(2)：246-248
34 Ito K, Ozasa H, Horikawa S. Edaravone protects against lung injury induced by intestiral ischemia/reperfusion in rat[J]. Free Radic Biol Med, 2005,38(3):369-374
35 Reyes YA, Shimoyama T. MCT-186(edaravone), a free radical scavenger, attennates ischemia-reperfusion injury and activation of phospholipase A(2) in an isolanted rat lung modEl after 18h of cold preserration[J]. Eur J Cardiothorac Surg,2006,29(3),304-311
36赵利军,董淑云.氧化应激在大鼠肢体缺血/再灌注心肌损伤中的作用[J].解放军医学杂志,2006,31(8)：802-803
37 Minhaz U,Tanaka M. Effect of MCI-186 on postischemic reperfusion injury in isolated rat[J]. Free Radic Res,1996,24(5):361
38周洪霞,张宇新,张连元,等.大鼠肢体缺血再灌注可致脑组织细胞凋亡[J]．解剖学杂志，2005,28(6):660-662
39周洪霞,张连元.大鼠肢体缺血再灌注脑组织Bax的表达变化及谷氨酸受体拮抗剂对其影响[J].华北煤炭医学院学报,2005,(7)2：142-143
40 Amemiya S, Kamiya T, Nito C, et al. Anti-apoptotic and neuroprotective effects of edaravone fol louing to unsient focalischemia in rats[J]. Eur J Pharmacol,2005,516(2):125-130
41 Yamamo T, Yuki S, Watonabe T, et al. DElayed neuronal death prevented by inhibition of increased hydroxyl radical formation in a transient cerebral ischemia[J]. Brain Res,1997,762(1-2):240-242
42王银环,董淑云.牛磺酸对大鼠肢体缺血再灌注后肝脏损伤的保护效应[J].第四军医大学学报,2006,27(22)：2047-2050
43 Tsvji K, Kwon AH, Yoshida H, et al. Free radical Scavenger (edaravone) prevents endotoxin-induced liver injury after partial hepatectomy in rats[J]. J Hepatol,2005,42(1):94-101