小肠缺血再灌注后远隔器官次级损伤的实验研究
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摘要
小肠缺血再灌注后可引起小肠组织一系列病理生理改变如血管内皮细胞肿胀和破坏，氧自由基的产生，微血管收缩，血小板及中性粒细胞粘附聚集，血流的降低等，其中氧自由基的改变，可激活相关酶系统及凋亡相关基因，诱导细胞凋亡，造成组织细胞功能和形态上的损伤。有研究表明小肠缺血再灌注除引起肠组织本身损伤外，还可引起远隔脏器的次级损伤。我们通过建立大鼠小肠缺血再灌注模型，对血中一氧化氮和超氧化物歧化酶的改变，心、肾、肝、肺组织中Bax，Bcl-2，P53的表达情况及电镜下各组织超微结构的变化进行研究，探讨小肠缺血再灌注损伤后对于远隔器官的次级损害，为研究各种原因引起的缺血再灌注造成远隔器官的次级损害的机制和防治建立基础。
研究目的 研究大鼠小肠缺血再灌注后血中NO、SOD的浓度变化，肺、肝、心、肾组织中Bax，Bcl-2，P53的表达以及电镜下超微结构的改变，探讨小肠缺血再灌注后对各远隔器官的损伤。
研究方法 建立大鼠小肠缺血再灌注模型，分对照组，再灌注后0、30min，1、2h，1、3、7d共8组，于各时点检测血中NO、SOD的浓度，用免疫组织化学SP法观察肺、肝、心、肾组织中Bax，Bcl-2，P53的表达，透射电镜下观察各组织超微结构的改变。
研究结果 大鼠小肠缺血再灌注后NO浓度0min明显升高，再灌注2h时降低，随后升高，再灌注7d时达高峰。SOD浓度再灌注0min明显下降，再灌注2h时升高，随后下降，再灌注7d时达最低水平。再灌注0min，肺、肝、心、肾组织中Bax，Bcl-2，p53阳性细胞率增多，再灌注30min时Bax，Bcl-2，p53阳性细胞率均升高，bcl-2表达高于Bax，两者差别显著(p＜0.01)。再灌注2h时Bax，Bcl-2，p53阳性细胞率降低，其后升高，再灌注7d时阳性细胞率达
第四军医大学硕士毕业论文
最高水平，Bax表达明显高于 bclZ，两者差别显著中＜0刀 1人透射电镜显示
各组织细胞染色质浓缩边集，线粒体异常等超微结构明显损伤改变。
结论 大鼠小肠缺血再灌注后血中NO，SOD的浓度明显变化、Bax，Bcl毛，
P53阳性细胞在心、肾、肝、肺组织中的表达的显著性改变以及各组织细胞超
微结构的改变说明小肠缺血再灌注后可引起远隔器官组织细胞次级损伤。
It might be a series of small intestin tissue pathophysiological processes such as swelling and destruction of vascular endothelial cell, prodution of oxygen-derived free radidicals, microvasoconstriction, assembling of platelet and neutrophil, reduction of blood flow after small intestine IR. The change of oxygen-derived free radidicals may active correlative enzyme system and apoptosis gene, induce cell apoptosis then result in morphologic tissue injuries. Researches have shown that small intestine IR not only injure bowl itself but also induce remote organ second injuries. Via building small intestine IR modles, we exame the concentration of NO and SOD in the blood, observe the expression of Bax, bcl-2, P53 and the change of ultrastructure in the remote organ of the lung, liver, heart and keydney to study IR-induced remote organ second injuries and to protect remote organ from IR-induced second injury for various reasons.
Abstract
AIM: To study the change of concentration of NO and SOD in the blood, the expression of Bax, bcl-2, P53, the change of ultrastructure in the remote organ of the lung, liver, heart and keydney and try to find out the small intestine IR-induced remote organ second injuries of the rat. The change of ultrastructure was observed by transmission electron microscopy.
METHODS: To make models of ischemia-reperfusion of small intestine at 0, 30min, 1, 2h, 1, 3, 7d after reperfusion, the concentration of NO, SOD in the blood was examed, the expression of Bax, bcl-2, Psa in the remote organ was observed by the immunohistochemical SP method. Remoe organ ultrastructures were observed by transmission electron microscopy.
RESULTS: The concentration of NO increased apparently after I/R 0 minute, but decreased significantly after I/R 2 hours, then increased gradually to a peak after 7 days. For SOD, the concentration decreased after I/R 0 minute, increased significantly after I/R 2 hours, and decreased gradually to the lowest level after I/R 7 days. The positive cells were observed. The ratio of positive cells of Bax, bcl-2, P53 increased after I/R 30 min, the ratio of positive cells of bcl-2 was higher than that of Bax (p<0.01) , after I/R two hours it decreased apparently, then increased after I/R seven days, and the ratio of Bax was higher than that of bcl-2 (p<0.01 ) .Remoe organ ultrastructures were serious injured observed by transmission electron microscopy such as chromatin concintration and mitochondria
disorder.
CONCLUSION: The change of concentration of NO and SOD in blood, the expression of positive cells of Bax, bcl-2, P53 as well as the change of Remoe organ ultrastructures show that it might cause apoptosis and injuries in the remote organ after IR of small intestine of rat.
研究目的 研究大鼠小肠缺血再灌注后血中NO、SOD的浓度变化，肺、肝、心、肾组织中Bax，Bcl-2，P53的表达以及电镜下超微结构的改变，探讨小肠缺血再灌注后对各远隔器官的损伤。
研究方法 建立大鼠小肠缺血再灌注模型，分对照组，再灌注后0、30min，1、2h，1、3、7d共8组，于各时点检测血中NO、SOD的浓度，用免疫组织化学SP法观察肺、肝、心、肾组织中Bax，Bcl-2，P53的表达，透射电镜下观察各组织超微结构的改变。
研究结果 大鼠小肠缺血再灌注后NO浓度0min明显升高，再灌注2h时降低，随后升高，再灌注7d时达高峰。SOD浓度再灌注0min明显下降，再灌注2h时升高，随后下降，再灌注7d时达最低水平。再灌注0min，肺、肝、心、肾组织中Bax，Bcl-2，p53阳性细胞率增多，再灌注30min时Bax，Bcl-2，p53阳性细胞率均升高，bcl-2表达高于Bax，两者差别显著(p＜0.01)。再灌注2h时Bax，Bcl-2，p53阳性细胞率降低，其后升高，再灌注7d时阳性细胞率达
第四军医大学硕士毕业论文
最高水平，Bax表达明显高于 bclZ，两者差别显著中＜0刀 1人透射电镜显示
各组织细胞染色质浓缩边集，线粒体异常等超微结构明显损伤改变。
结论 大鼠小肠缺血再灌注后血中NO，SOD的浓度明显变化、Bax，Bcl毛，
P53阳性细胞在心、肾、肝、肺组织中的表达的显著性改变以及各组织细胞超
微结构的改变说明小肠缺血再灌注后可引起远隔器官组织细胞次级损伤。
It might be a series of small intestin tissue pathophysiological processes such as swelling and destruction of vascular endothelial cell, prodution of oxygen-derived free radidicals, microvasoconstriction, assembling of platelet and neutrophil, reduction of blood flow after small intestine IR. The change of oxygen-derived free radidicals may active correlative enzyme system and apoptosis gene, induce cell apoptosis then result in morphologic tissue injuries. Researches have shown that small intestine IR not only injure bowl itself but also induce remote organ second injuries. Via building small intestine IR modles, we exame the concentration of NO and SOD in the blood, observe the expression of Bax, bcl-2, P53 and the change of ultrastructure in the remote organ of the lung, liver, heart and keydney to study IR-induced remote organ second injuries and to protect remote organ from IR-induced second injury for various reasons.
Abstract
AIM: To study the change of concentration of NO and SOD in the blood, the expression of Bax, bcl-2, P53, the change of ultrastructure in the remote organ of the lung, liver, heart and keydney and try to find out the small intestine IR-induced remote organ second injuries of the rat. The change of ultrastructure was observed by transmission electron microscopy.
METHODS: To make models of ischemia-reperfusion of small intestine at 0, 30min, 1, 2h, 1, 3, 7d after reperfusion, the concentration of NO, SOD in the blood was examed, the expression of Bax, bcl-2, Psa in the remote organ was observed by the immunohistochemical SP method. Remoe organ ultrastructures were observed by transmission electron microscopy.
RESULTS: The concentration of NO increased apparently after I/R 0 minute, but decreased significantly after I/R 2 hours, then increased gradually to a peak after 7 days. For SOD, the concentration decreased after I/R 0 minute, increased significantly after I/R 2 hours, and decreased gradually to the lowest level after I/R 7 days. The positive cells were observed. The ratio of positive cells of Bax, bcl-2, P53 increased after I/R 30 min, the ratio of positive cells of bcl-2 was higher than that of Bax (p<0.01) , after I/R two hours it decreased apparently, then increased after I/R seven days, and the ratio of Bax was higher than that of bcl-2 (p<0.01 ) .Remoe organ ultrastructures were serious injured observed by transmission electron microscopy such as chromatin concintration and mitochondria
disorder.
CONCLUSION: The change of concentration of NO and SOD in blood, the expression of positive cells of Bax, bcl-2, P53 as well as the change of Remoe organ ultrastructures show that it might cause apoptosis and injuries in the remote organ after IR of small intestine of rat.
引文
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