止血带压迫对大鼠坐骨神经轴浆转运的影响
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摘要
背景：止血带用于肢体手术,可以减少病人出血量,清晰手术视野,便于解剖结构辨认,缩短手术时间,减少并发症,目前广泛应用于骨科和整形外科手术。但应用止血带压迫本身也可以引起一些不良反应,如出现疼痛、高血压等。既往对止血带导致不良反应的研究主要集中在止血带压迫区域,或者是止血带压迫下方的缺血区域,而对于止血带压迫上方区域研究较少。本实验通过观察止血带压迫大鼠坐骨神经对类胰岛素样生长因子-1(Insulin-like growth factor-1, IGF-1)运输的变化情况,来研究止血带压迫对轴浆转运的影响。
方法：采用12周龄的雄性sprague dawley大鼠,48只,体重250-300g,随机分为8组：止血带压迫1h组(A组)、2h组(B组)、4h组(C组)和12h组(D组)、神经结扎1h组(E组)、2h组(F组)、4h组(G组)和12h组(H组),每组6只。全身麻醉后,随机决定每只大鼠的左侧或右侧坐骨神经是处理组或对照组。压迫或结扎达到目标时间后取材,通过免疫组化的方法观察坐骨神经受压或结扎部位上方、下方IGF-1的堆积情况,然后计算出每张免疫组化图片的平均光密度值,用处理组平均光密度值与对照组平均光密度值之比(ratio of experimental average optic density to control, E/C)来判断IGF-1堆积的程度。
结果：D组有一只大鼠死亡,其实验数据予以排除,其余四十七只大鼠实验结果纳入分析使用。A、B、C、D组为止血带压迫组,止血带压迫上方区域有明显的IGF-1免疫反应性物质堆积,E/C值均大于1,随着压迫时间的延长而增加,分别为1.30±0.28、1.57±0.30、2.78±0.56、5.91±1.22。止血带压迫下方区域未见明显IGF-1免疫反应性物质堆积,E/C值在1左右,也不随压迫时间的延长而改变,分别为1.03±0.05、1.00±0.01、1.08±0.12、1.02±0.04。A组内的三只大鼠,B组内的一只大鼠坐骨神经压迫部位上方区域IGF-1免疫反应性物质堆积不明显；E、F、G、H组为神经结扎组,结扎痕迹上方及下方都有IGF-1免疫反应性物质的明显堆积,且结扎痕迹上方区域堆积的程度较下方明显。四组在结扎痕迹上方及下方区域E/C值都大于1,且随着时间的延长而增加。结扎上方的E/G值分别为1.53±0.51、2.89±0.67、4.04±0.77和7.99±1.18。结扎下方的E/G值分别为1.19±0.24、1.86±0.24、2.17±0.33和4.04±1.69。E组有两只大鼠坐骨神经结扎部位上方及下方IGF-1免疫反应性物质堆积不明显。
结论：1.止血带压迫可影响坐骨神经的轴浆转运,在止血带压迫部位的上方区域出现IGF-1的明显堆积,并随时间延长增加；而止血带压迫部位的下方区域未见明显IGF-1堆积,也不随时间改变。2.结扎大鼠坐骨神经可以阻断坐骨神经的轴浆转运,在结扎部位的上方和下方均出现明显的IGF-1堆积,并随时间延长增加；
3.神经结扎导致的IGF-1堆积程度明显超过止血带压迫导致的IGF-1堆积。
Background:Tourniquet is widely useu in extremity surgery to reduce bleeding, achieve a bloodless field and facilitate the identification of anatomical structures, which is expected to shorten the operative time and decrease complications. So the tourniquet is extensive used in orthopedic and plastic surgery. It also can cause some pathophysiology changes, even adverse reactions, for example tourniquet pain, tourniquet hypertension and so on. The research of tourniquet side effect mainly focus on the area, which is compressed by tourniquet or distal to compressed site. But few research pay attention to the area, which is proximal tourniquet compressed site. In this experiment, to study the tourniquet compression on axonal transport, the transport of insulin like growth factor-1 in rat sciatic nerve axon was observed.
Methods:Forty-eight 12-week-old, weight 250-300g male sprague dawley rats were included in this study, and random divided into eight groups:Tourniquet compressed 1h group (group A),2h group (group B),4h group (group C),12h group (group D), nerve ligation 1h group (group E),2h group (group F),4h group (group G),12h group (group H). Six rats in every group. After anesthesia, the left or right side of each rat sciatic nerve is randomly separated into the treatment group or control group. After treatment reach target time, drawing materials. And, the immunohistochemistry was used to observation the accumulation of IGF-1 of the area around sciatic nerve compression or ligation site. Then calculated the average optical density for each immunohistochemical picture. After that the ratio of experimental average optic density to control was used to measure the degree of IGF-1 accumulation.
Results:One of rats was dead in group D, and the data was excluded from the result. The experimental data of remaining 47 rats were included in the analysis. Group A, B, C and D was tourniquet compression group, there was obvious IGF-1 immunoreactivity material accumulation on the area proximal tourniquet compressed site. And all of the E/C values were greater than 1. The accumulation was severer with the compressed time extended. The E/C values of group A, B, C and D were 1.30±0.28,1.57±0.30,2.78 ±0.56,5.91±1.22, respectively. But there was no significant IGF-1 immunoreactivity material accumulation on the area distal to tourniquet compressed site. And the values of group A, B, C, D were 1.03±0.05,1.00±0.01,1.08±0.12,1.02±0.04, around 1, there was no obvious change with treatment time extended. There were three rats in group A and one rat in group B no obvious IGF-1 immunoreactivity material accumulation at the proximal Sciatic nerve compression site. Group E, F, G and H were nerve ligation group. Both proximal and distal compressed site were IGF-1 immunoreactivity material accumulation, and the accumulation of proximal was more severe. The values of group E, F, G, H were 1.53±0.51,2.89±0.67,4.04±0.77,7.99±1.18 at the proximal ligation site,1.19±0.24、1.86±0.24、2.17±0.33和4.04±1.69 at the distal ligation site. There were two rats in group E no obvious IGF-1 immunoreactivity material accumulation.
Conclusion:1.The use of tourniquet can affect the sciatic nerve axonal transport. There were apparent accumulation of IGF-1 in proximal tourniquet compressed site, and accumulation degree severer with time extended. But in distal tourniquet compressed site, there were no significant accumulation, and little change with time extended. 2. The ligation of rat sciatic nerve can block axonal transport, both the proximal and distal ligation site were apparent IGF-1 accumulation, and severer with time extended. 3. The accumulation degree of IGF-1 was severer in nerve ligation group than tourniquet compressed group.
方法：采用12周龄的雄性sprague dawley大鼠,48只,体重250-300g,随机分为8组：止血带压迫1h组(A组)、2h组(B组)、4h组(C组)和12h组(D组)、神经结扎1h组(E组)、2h组(F组)、4h组(G组)和12h组(H组),每组6只。全身麻醉后,随机决定每只大鼠的左侧或右侧坐骨神经是处理组或对照组。压迫或结扎达到目标时间后取材,通过免疫组化的方法观察坐骨神经受压或结扎部位上方、下方IGF-1的堆积情况,然后计算出每张免疫组化图片的平均光密度值,用处理组平均光密度值与对照组平均光密度值之比(ratio of experimental average optic density to control, E/C)来判断IGF-1堆积的程度。
结果：D组有一只大鼠死亡,其实验数据予以排除,其余四十七只大鼠实验结果纳入分析使用。A、B、C、D组为止血带压迫组,止血带压迫上方区域有明显的IGF-1免疫反应性物质堆积,E/C值均大于1,随着压迫时间的延长而增加,分别为1.30±0.28、1.57±0.30、2.78±0.56、5.91±1.22。止血带压迫下方区域未见明显IGF-1免疫反应性物质堆积,E/C值在1左右,也不随压迫时间的延长而改变,分别为1.03±0.05、1.00±0.01、1.08±0.12、1.02±0.04。A组内的三只大鼠,B组内的一只大鼠坐骨神经压迫部位上方区域IGF-1免疫反应性物质堆积不明显；E、F、G、H组为神经结扎组,结扎痕迹上方及下方都有IGF-1免疫反应性物质的明显堆积,且结扎痕迹上方区域堆积的程度较下方明显。四组在结扎痕迹上方及下方区域E/C值都大于1,且随着时间的延长而增加。结扎上方的E/G值分别为1.53±0.51、2.89±0.67、4.04±0.77和7.99±1.18。结扎下方的E/G值分别为1.19±0.24、1.86±0.24、2.17±0.33和4.04±1.69。E组有两只大鼠坐骨神经结扎部位上方及下方IGF-1免疫反应性物质堆积不明显。
结论：1.止血带压迫可影响坐骨神经的轴浆转运,在止血带压迫部位的上方区域出现IGF-1的明显堆积,并随时间延长增加；而止血带压迫部位的下方区域未见明显IGF-1堆积,也不随时间改变。2.结扎大鼠坐骨神经可以阻断坐骨神经的轴浆转运,在结扎部位的上方和下方均出现明显的IGF-1堆积,并随时间延长增加；
3.神经结扎导致的IGF-1堆积程度明显超过止血带压迫导致的IGF-1堆积。
Background:Tourniquet is widely useu in extremity surgery to reduce bleeding, achieve a bloodless field and facilitate the identification of anatomical structures, which is expected to shorten the operative time and decrease complications. So the tourniquet is extensive used in orthopedic and plastic surgery. It also can cause some pathophysiology changes, even adverse reactions, for example tourniquet pain, tourniquet hypertension and so on. The research of tourniquet side effect mainly focus on the area, which is compressed by tourniquet or distal to compressed site. But few research pay attention to the area, which is proximal tourniquet compressed site. In this experiment, to study the tourniquet compression on axonal transport, the transport of insulin like growth factor-1 in rat sciatic nerve axon was observed.
Methods:Forty-eight 12-week-old, weight 250-300g male sprague dawley rats were included in this study, and random divided into eight groups:Tourniquet compressed 1h group (group A),2h group (group B),4h group (group C),12h group (group D), nerve ligation 1h group (group E),2h group (group F),4h group (group G),12h group (group H). Six rats in every group. After anesthesia, the left or right side of each rat sciatic nerve is randomly separated into the treatment group or control group. After treatment reach target time, drawing materials. And, the immunohistochemistry was used to observation the accumulation of IGF-1 of the area around sciatic nerve compression or ligation site. Then calculated the average optical density for each immunohistochemical picture. After that the ratio of experimental average optic density to control was used to measure the degree of IGF-1 accumulation.
Results:One of rats was dead in group D, and the data was excluded from the result. The experimental data of remaining 47 rats were included in the analysis. Group A, B, C and D was tourniquet compression group, there was obvious IGF-1 immunoreactivity material accumulation on the area proximal tourniquet compressed site. And all of the E/C values were greater than 1. The accumulation was severer with the compressed time extended. The E/C values of group A, B, C and D were 1.30±0.28,1.57±0.30,2.78 ±0.56,5.91±1.22, respectively. But there was no significant IGF-1 immunoreactivity material accumulation on the area distal to tourniquet compressed site. And the values of group A, B, C, D were 1.03±0.05,1.00±0.01,1.08±0.12,1.02±0.04, around 1, there was no obvious change with treatment time extended. There were three rats in group A and one rat in group B no obvious IGF-1 immunoreactivity material accumulation at the proximal Sciatic nerve compression site. Group E, F, G and H were nerve ligation group. Both proximal and distal compressed site were IGF-1 immunoreactivity material accumulation, and the accumulation of proximal was more severe. The values of group E, F, G, H were 1.53±0.51,2.89±0.67,4.04±0.77,7.99±1.18 at the proximal ligation site,1.19±0.24、1.86±0.24、2.17±0.33和4.04±1.69 at the distal ligation site. There were two rats in group E no obvious IGF-1 immunoreactivity material accumulation.
Conclusion:1.The use of tourniquet can affect the sciatic nerve axonal transport. There were apparent accumulation of IGF-1 in proximal tourniquet compressed site, and accumulation degree severer with time extended. But in distal tourniquet compressed site, there were no significant accumulation, and little change with time extended. 2. The ligation of rat sciatic nerve can block axonal transport, both the proximal and distal ligation site were apparent IGF-1 accumulation, and severer with time extended. 3. The accumulation degree of IGF-1 was severer in nerve ligation group than tourniquet compressed group.
引文
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