脊髓损伤后脊髓组织内凝血酶受体1的表达

Expression of thrombin receptor-1 in spinal cord tissue after spinal cord injury

目的:观察脊髓损伤后脊髓组织内凝血酶受体1的表达,分析表达规律与脊髓损伤程度是否具有时效性。方法:①实验于2005-04/08在兰州大学基础医学院解剖学教研室实验室和甘肃省医学科学研究院动物实验中心完成。②选用成年健康Wistar大鼠75只,雌雄不拘。按随机抽签法将大鼠分为3组:正常对照组:只打开椎板,不损伤脊髓;模型组:打开椎板,损伤脊髓;脊髓损伤+注全血组:打开椎板,损伤脊髓,立即自大鼠尾静脉取20μL自体血,注入骨窗中心的损伤脊髓组织中,缓慢注射,注射完毕后留置3min。③于术后6,24,48,72h和5d,采用免疫组织化学法检测各组脊髓组织凝血酶受体1表达。电镜观察各组术后72h神经元的超微结构变化。苏木精-伊红染色后对脊髓组织进行形态学观察。结果:大鼠75只均进入结果分析。①凝血酶受体1阳性表达神经元数:模型组和脊髓损伤+注全血组明显多于正常对照组(P<0.01);脊髓损伤+注全血组明显多于模型组(P<0.01)。模型组和脊髓损伤+注全血组脊髓组织阳性细胞表达上调开始于术后6h(每400倍视野阳性细胞数分别为6.20±1.48,8.30±1.42),逐渐升高,72h达高峰(每400倍视野阳性细胞数分别为14.50±1.43,20.40±1.71),以后逐渐降低。②脊髓组织形态学观察结果:术后6h模型组和脊髓损伤+注全血组神经细胞出现轻度水肿,术后72h神经细胞和细胞周围水肿非常明显,之后水肿情况逐渐减轻;脊髓损伤+注全血组脊髓组织水肿较模型组重。③术后72h各组脊髓组织超微结构:模型组和脊髓损伤+注全血组脊髓组织出现神经元凋亡。正常对照组见正常神经元。结论:脊髓损伤后脊髓组织中神经元凝血酶受体1表达增强,其表达规律与脊髓损伤程度有一定的时效关系,凝血酶受体1的表达可能参与了脊髓组织继发性损伤。

AIM： To detect the expression of thrombin receptor-1 （PAR-1） after spinal cord injury, and investigate whether there is a time-effect relationship between expressive rule and injured degree. METHODS： （1） The experiment was conducted in .the laboratory of Department of Human Anatomy, School of Basic Medicine, Lanzhou University and the animal experimental center of Gansu Medical Research College from April to August 2005. （2）Seventy-five healthy adult Wistar rats were randomly divided into 3 groups： control group, only the neural scute was opened; model group, only the neural scute was opened and the spinal cord was injured; spinal cord injury plus whole blood injection group （SCI-B） the neural scute was opened and the spinal cord was injured and 20 μL whole blood was slowly injected into the injured tissue, and detained for 3 minutes. （3） Six hours, twenty-four hours, forty-eight hours, seventy-two hours and five days after operation, the spinal cord tissue was taken out to detect the PAR-1 expression by immunohistochemical method. The changes of neuron uhrastructure were observed by electron microscope at seventy-two hours and the morph changes were observed after HE staining. RESULTS： All the 75 rats were involved in the result analysis. （1）Number of PAR-l-positive cells： The model group and SCI-B group were markedly more than the control group （P 〈 0.01）, the SCI-B group was markedly increased compared with model group （P 〈 0.01）. The expression of PAR-1 was found in the model group and SCI-B group at six hours after Operation （6.20±1.48, 8.30±1.42 per 400-fold field）, and gradually increased, after reached the peak at 72 hours （14.50±1.43, 20.40±1.71 per 400-fold field）, it began to decrease gradually. （2） Morphological observation of spinal tissue： Slight edema of spinal cord tissue was found in the model and SCI-B groups at 6 hours after operation, which became more serious at 72 hours, and then gradually improved; the edema in the SCI-B group was more serious than the model group. （3）Uhrastructure of the spinal tissue after 72 hours： Neural apoptosis was found in the model and SCI-B groups; but normal neurons were observed in the normal control group. CONCLUSION： The expression of PAR-I in spinal cord tissue is upregulated after SCI. There is a time-effect relationship between the expressive rule and injured degree of spinal cord, and PAR-1 expression might be involved in neuron injuries after SCI.