兔骶丛爆震伤动物模型的建立及相关性实验研究
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摘要
盆骶部火器伤是现代战伤的一个重要部位,由于解剖结构复杂,生理功能特殊,合并伤及并发症多,具有高感染率、高致残率、高死亡率的特点,是战创伤救治的难题和重点。骶丛战伤损伤范围广泛,大多不能一期修复,加重了失神经支配器官的功能障碍。现代战争更加侧重于爆炸性武器的应用,因此,建立骶丛爆震伤模型,揭示神经变性及靶细胞的功能退变规律,为骶丛火器伤的救治提供一定的理论依据,具有非常重要的军事及医学意义,以最大程度减低战场官兵的致残率。
目的1、研究兔骶丛的组成及其支配器官。
2、建立兔骶丛爆震伤动物模型。
3、通过兔骶丛爆震伤动物模型,观察骶丛神经损伤段、骶丛损伤远段(坐骨神经段)、中枢神经元凋亡情况、靶肌肉失神经萎缩情况。
方法1、新鲜大耳白兔标本10只,共20侧。从前后路切开,从腰椎椎间孔与骶椎骶孔开始,尖刀锐性性分离神经与周围组织,从近端分离至远端,探查神经至靶器官及肌肉的起止点。
2、大耳白兔18只,致伤物为当量600mgTNT炸药的电子雷管,根据雷管与皮肤致伤距离不同随机分为致伤物与皮肤距离为1cm(A)、致伤物与皮肤距离为2cm(B)、致伤物与皮肤距离为5cm(C)三组,以后正中线、右髂后上棘与股骨粗隆连线、右股骨粗隆水平连线形成—三角区域,即骶丛在体表投影区域为致伤点,根据下肢体感诱发电位致伤前、致伤即刻、致伤1周后3个时相点检测及动物的伤情评估,来观察骶丛神经损伤的情况。
3、大耳白兔36只,随机分为正常组(A)、1d组(B)、3d组(C)、1w周(D)、2w组(E)、4w组(F)等6组。致伤物为当量600mgTNT炸药的电子雷管,雷管与皮肤致伤距离为2cm,以后正中线、右髂后上棘与股骨粗隆连线、右股骨粗隆水平连线形成一三角区域,即骶丛在体表投影区域为致伤点,根据下肢体感诱发电位致伤后检测明显消失及动物的出现骶丛神经损伤表现,纳入实验组。通过HE染色、TUNEL检测、电镜观察,光镜下观察骶丛、骶髓、腓肠肌组织,神经手工计数神经元细胞。
结果1、骶丛主要由腰5～7神经干及骶神经组成,其神经分支有臀前神经、坐骨神经、股后皮神经、阴部神经、.直肠中神经、直肠后神经；靶器官及肌肉支配有直肠、泌尿生殖道、臀大肌、臀中肌、梨状肌、股二头肌、半腱肌、半膜肌、小腿三头肌、胫前肌、腓骨长肌、腓骨短肌、第三腓骨肌、第四腓骨肌、趾长伸肌、踇长伸肌、趾浅屈肌、趾长屈肌、骨间肌、蚓状肌。
2、A组肉眼下骶丛神经全部断裂,1周存活率为0；B组肉眼下骶丛神经挫伤,骶丛损伤症状明显,伤后即刻SLSEP消失,1周后无恢复,1周存活率83%；C组仅伤及皮肤,未见骶丛损伤症状,伤后即刻SLSEP轻度改变(p<0.05),1周后与伤前比较无明显差别(p>0.05),1周存活率100%。
3、爆震伤后骶丛神经直接损伤段1～3天主要表现为束膜内出血,2周后神经轴突内出现明显瘢痕增生。1d～3d时脊髓伤侧前角神经元计数与正常组比较,差异不显著(P>0.05)；1周时神经元数与正常组相比显著降低(P<0.01)。爆震伤后1d即可见到脊髓凋亡细胞,伤后第3d为凋亡发生的高峰期,1周组与3d组无明显差别,与正常组和其他组相比显著增多(P<0.01)。靶肌肉(腓肠肌)在2周后出现明显失神经萎缩。
结论1、兔的骶神经的解剖位置、神经干的组成及靶器官的支配,与人的骶丛大致相似,可作为关于骶丛方面实验动物选择。
2、B组600mgTNT炸药,致伤距离2cm,致伤压力51.34±1.03MPa,可形成典型的骶丛损伤,稳定性良好,且存活率高,可作为长期实验观察研究。
3、骶丛爆震伤神经段损伤广泛严重,中枢脊髓神经元凋亡出现早,靶肌肉出现失神经萎缩,因此骶丛神经爆震损伤需要更早的进行治疗。
Pelvis Sacral firearm injury is an important part of modern war injuries. Because of complicated anatomical, special physiological functions, more associated injury and complications,there are have high rate of infection,mutilation and mortality.so, it is a tough problem and emphasis of firearm injury treatment. The range of Sacral plexus firearm injury is wide, but almost can't primary repair. So the denervated organs aggravate dysfunction. Modern warfare is more focused on the application of explosive weapons, thus, it's very important significance in military and medical to establish the model of the Sacral Plexus explosion Injury, revealed the laws of neural degeneration and target cells degeneration, for provide the treatment theories of sacral plexus firearm injury and to maximum reduce the disability of injured soldiers.
Objective 1、To consider composition and dominance of Plexus and organ in rabbits.
2、To establish the model of the Sacral Plexus explosion Injury in Rabbits.
3、To observe the changes of Sacral Plexus'direct segment, distal segment(Sciatic nerve), spinal cord neuronal apoptosis, and denervated muscle atrophy following explosion injury of Sacral Plexus in rabbits.
Methods 1、There are 10 fresh specimens of rabbits, a total of 20 sides. The incision conclude both previous and posterior. Separated nerves and tissue start from lumbar vertebra foramina and sacral vertebra foramina by knife. Separated from the proximal to distal direction. To detect the beginning and ending of nerves and muscles.
2、18 Rabbits were divided into A,B,C groups at random which were injured by electronic explosive detonators contain 600mg TNT. The distance between detonator and skin was lcm in A group,2cm in B group,5cm in C group. The hit point was a triangle formed by posterior median line, the right posterior superior iliac spine with femur connection line, and the level line of the right femur, where was the surface projection of the Sacral Plexus region. The Sacral Plexus nerve injury situation was observed according to record animal injury assessment and short-latency somatosensory evoked potential (SLSEP) at the time of before injury, immediately after injury and 1 w after injury.
3、6 rabbits were divided into the control group(A),1d group(B),3d group(C),1w group(D),2w group(E),4w(F) group at random which were injured by electronic explosive detonators contain 600mg TNT. The distance between detonator and skin was 2 cm. The hit point was a triangle formed by posterior median line, the right posterior superior iliac spine with femur connection line, and the level line of the right femur, where was the surface projection of the Sacral Plexus region. The accepted ones were according to short-latency somatosensory evoked potential (SLSEP) disappeared and the Sacral Plexus nerve injuried situation was observed after injuried. According to HE stain,TUNEL and electron microscope,to observe Sacral plexus, sacral and gastrocnemius, to record the number of neurons and neuronal apoptosis by manual counted.
Results 1、The sacral nerves were mainly composed of 5-7 lumbar nerves and all anterior branch sacral nerves. The nervous ramification conclude nerves glutaeus cranialis, sciatic nerve, posterior femoral cutaneous nerve, pudendal nerve, meso-rectum nerve, aboral-rectum nerve. The target organs and muscles concluded rectum, appendage urogenitalis, gluteus maximus muscle, gluteus medius muscle, piriform muscle, biceps flexor cruris,semitendinosus,semimembranous muscle, triceps surae muscle, anterior tibial muscle, long fibular muscle, peroneous brevis, peroneus tertius, forth fibular muscle, extensor digitorum longus, extensor pollicis longus, flexor digitorum superficialis muscle,flexor digitorum longus muscle, interosseus,lumbricales.
2、The Sacral Plexus were all rupture, survival rate of 1 w was 0 in A group. The Sacral Plexus were bruised in B group and occurred obvious injury symptoms. The SLSEP disappeared immediately after injury,and didn't recover after 1 w, survival rate of 1w was 83%. The skin was hurt but no Sacral Plexus injury symptoms in C group. The SLSEP little changed immediately after injury (p<0.05), but recovered after 1 w compared before injury with no significant difference (p> 0.05), survival rate of 1w was 100%.
3、The mainly changes of Sacral Plexus'direct segments were hemorrhage in intraperineurium in Id to 3d. There were appearance scar in Axons after 2 weeks. There Was no significant differences in anterior horn neurons of spinal cord of the injured side 1d-3d after injury in the injury group and the control group(P>0.05), but the neuron number were decreased 1 weeks later in the injury group as compared with that of the control group(P<0.05). Apoptotic neurons were observed in spinal cord in Id group. The peak of apoptosis occurred 3 days after injury, which was similar to 1w group, but had significantly more than the control group and other groups. Gastrocnemius denervated muscle atrophy occurred after 2 weeks.
Conclusion 1、The Sacral Plexus anatomy,neural stem composition and the dominant target organs of rabbits are similar to human. Rabbits can be used as experimental animals in sacral plexus study.
2、Under the condition of 600mgTNT,2 cm detonator distance,and 51.34±1.03MPa blast pressure in B group,typical Sacral Plexus explosion injury could be gained easily, and the long time pathological changes of injured nerve could be observed.
3、The Sacral Plexus explosion injury are widespread and serious. Spinal cord neuronal apoptosis occurred early. Target muscles occurred denervated muscle atrophy. Therefore,the sacral plexus explosion injury need treatment earlier.
目的1、研究兔骶丛的组成及其支配器官。
2、建立兔骶丛爆震伤动物模型。
3、通过兔骶丛爆震伤动物模型,观察骶丛神经损伤段、骶丛损伤远段(坐骨神经段)、中枢神经元凋亡情况、靶肌肉失神经萎缩情况。
方法1、新鲜大耳白兔标本10只,共20侧。从前后路切开,从腰椎椎间孔与骶椎骶孔开始,尖刀锐性性分离神经与周围组织,从近端分离至远端,探查神经至靶器官及肌肉的起止点。
2、大耳白兔18只,致伤物为当量600mgTNT炸药的电子雷管,根据雷管与皮肤致伤距离不同随机分为致伤物与皮肤距离为1cm(A)、致伤物与皮肤距离为2cm(B)、致伤物与皮肤距离为5cm(C)三组,以后正中线、右髂后上棘与股骨粗隆连线、右股骨粗隆水平连线形成—三角区域,即骶丛在体表投影区域为致伤点,根据下肢体感诱发电位致伤前、致伤即刻、致伤1周后3个时相点检测及动物的伤情评估,来观察骶丛神经损伤的情况。
3、大耳白兔36只,随机分为正常组(A)、1d组(B)、3d组(C)、1w周(D)、2w组(E)、4w组(F)等6组。致伤物为当量600mgTNT炸药的电子雷管,雷管与皮肤致伤距离为2cm,以后正中线、右髂后上棘与股骨粗隆连线、右股骨粗隆水平连线形成一三角区域,即骶丛在体表投影区域为致伤点,根据下肢体感诱发电位致伤后检测明显消失及动物的出现骶丛神经损伤表现,纳入实验组。通过HE染色、TUNEL检测、电镜观察,光镜下观察骶丛、骶髓、腓肠肌组织,神经手工计数神经元细胞。
结果1、骶丛主要由腰5～7神经干及骶神经组成,其神经分支有臀前神经、坐骨神经、股后皮神经、阴部神经、.直肠中神经、直肠后神经；靶器官及肌肉支配有直肠、泌尿生殖道、臀大肌、臀中肌、梨状肌、股二头肌、半腱肌、半膜肌、小腿三头肌、胫前肌、腓骨长肌、腓骨短肌、第三腓骨肌、第四腓骨肌、趾长伸肌、踇长伸肌、趾浅屈肌、趾长屈肌、骨间肌、蚓状肌。
2、A组肉眼下骶丛神经全部断裂,1周存活率为0；B组肉眼下骶丛神经挫伤,骶丛损伤症状明显,伤后即刻SLSEP消失,1周后无恢复,1周存活率83%；C组仅伤及皮肤,未见骶丛损伤症状,伤后即刻SLSEP轻度改变(p<0.05),1周后与伤前比较无明显差别(p>0.05),1周存活率100%。
3、爆震伤后骶丛神经直接损伤段1～3天主要表现为束膜内出血,2周后神经轴突内出现明显瘢痕增生。1d～3d时脊髓伤侧前角神经元计数与正常组比较,差异不显著(P>0.05)；1周时神经元数与正常组相比显著降低(P<0.01)。爆震伤后1d即可见到脊髓凋亡细胞,伤后第3d为凋亡发生的高峰期,1周组与3d组无明显差别,与正常组和其他组相比显著增多(P<0.01)。靶肌肉(腓肠肌)在2周后出现明显失神经萎缩。
结论1、兔的骶神经的解剖位置、神经干的组成及靶器官的支配,与人的骶丛大致相似,可作为关于骶丛方面实验动物选择。
2、B组600mgTNT炸药,致伤距离2cm,致伤压力51.34±1.03MPa,可形成典型的骶丛损伤,稳定性良好,且存活率高,可作为长期实验观察研究。
3、骶丛爆震伤神经段损伤广泛严重,中枢脊髓神经元凋亡出现早,靶肌肉出现失神经萎缩,因此骶丛神经爆震损伤需要更早的进行治疗。
Pelvis Sacral firearm injury is an important part of modern war injuries. Because of complicated anatomical, special physiological functions, more associated injury and complications,there are have high rate of infection,mutilation and mortality.so, it is a tough problem and emphasis of firearm injury treatment. The range of Sacral plexus firearm injury is wide, but almost can't primary repair. So the denervated organs aggravate dysfunction. Modern warfare is more focused on the application of explosive weapons, thus, it's very important significance in military and medical to establish the model of the Sacral Plexus explosion Injury, revealed the laws of neural degeneration and target cells degeneration, for provide the treatment theories of sacral plexus firearm injury and to maximum reduce the disability of injured soldiers.
Objective 1、To consider composition and dominance of Plexus and organ in rabbits.
2、To establish the model of the Sacral Plexus explosion Injury in Rabbits.
3、To observe the changes of Sacral Plexus'direct segment, distal segment(Sciatic nerve), spinal cord neuronal apoptosis, and denervated muscle atrophy following explosion injury of Sacral Plexus in rabbits.
Methods 1、There are 10 fresh specimens of rabbits, a total of 20 sides. The incision conclude both previous and posterior. Separated nerves and tissue start from lumbar vertebra foramina and sacral vertebra foramina by knife. Separated from the proximal to distal direction. To detect the beginning and ending of nerves and muscles.
2、18 Rabbits were divided into A,B,C groups at random which were injured by electronic explosive detonators contain 600mg TNT. The distance between detonator and skin was lcm in A group,2cm in B group,5cm in C group. The hit point was a triangle formed by posterior median line, the right posterior superior iliac spine with femur connection line, and the level line of the right femur, where was the surface projection of the Sacral Plexus region. The Sacral Plexus nerve injury situation was observed according to record animal injury assessment and short-latency somatosensory evoked potential (SLSEP) at the time of before injury, immediately after injury and 1 w after injury.
3、6 rabbits were divided into the control group(A),1d group(B),3d group(C),1w group(D),2w group(E),4w(F) group at random which were injured by electronic explosive detonators contain 600mg TNT. The distance between detonator and skin was 2 cm. The hit point was a triangle formed by posterior median line, the right posterior superior iliac spine with femur connection line, and the level line of the right femur, where was the surface projection of the Sacral Plexus region. The accepted ones were according to short-latency somatosensory evoked potential (SLSEP) disappeared and the Sacral Plexus nerve injuried situation was observed after injuried. According to HE stain,TUNEL and electron microscope,to observe Sacral plexus, sacral and gastrocnemius, to record the number of neurons and neuronal apoptosis by manual counted.
Results 1、The sacral nerves were mainly composed of 5-7 lumbar nerves and all anterior branch sacral nerves. The nervous ramification conclude nerves glutaeus cranialis, sciatic nerve, posterior femoral cutaneous nerve, pudendal nerve, meso-rectum nerve, aboral-rectum nerve. The target organs and muscles concluded rectum, appendage urogenitalis, gluteus maximus muscle, gluteus medius muscle, piriform muscle, biceps flexor cruris,semitendinosus,semimembranous muscle, triceps surae muscle, anterior tibial muscle, long fibular muscle, peroneous brevis, peroneus tertius, forth fibular muscle, extensor digitorum longus, extensor pollicis longus, flexor digitorum superficialis muscle,flexor digitorum longus muscle, interosseus,lumbricales.
2、The Sacral Plexus were all rupture, survival rate of 1 w was 0 in A group. The Sacral Plexus were bruised in B group and occurred obvious injury symptoms. The SLSEP disappeared immediately after injury,and didn't recover after 1 w, survival rate of 1w was 83%. The skin was hurt but no Sacral Plexus injury symptoms in C group. The SLSEP little changed immediately after injury (p<0.05), but recovered after 1 w compared before injury with no significant difference (p> 0.05), survival rate of 1w was 100%.
3、The mainly changes of Sacral Plexus'direct segments were hemorrhage in intraperineurium in Id to 3d. There were appearance scar in Axons after 2 weeks. There Was no significant differences in anterior horn neurons of spinal cord of the injured side 1d-3d after injury in the injury group and the control group(P>0.05), but the neuron number were decreased 1 weeks later in the injury group as compared with that of the control group(P<0.05). Apoptotic neurons were observed in spinal cord in Id group. The peak of apoptosis occurred 3 days after injury, which was similar to 1w group, but had significantly more than the control group and other groups. Gastrocnemius denervated muscle atrophy occurred after 2 weeks.
Conclusion 1、The Sacral Plexus anatomy,neural stem composition and the dominant target organs of rabbits are similar to human. Rabbits can be used as experimental animals in sacral plexus study.
2、Under the condition of 600mgTNT,2 cm detonator distance,and 51.34±1.03MPa blast pressure in B group,typical Sacral Plexus explosion injury could be gained easily, and the long time pathological changes of injured nerve could be observed.
3、The Sacral Plexus explosion injury are widespread and serious. Spinal cord neuronal apoptosis occurred early. Target muscles occurred denervated muscle atrophy. Therefore,the sacral plexus explosion injury need treatment earlier.
引文
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