CAJ | 학술논문

目的探索失血性休克对肠黏膜细胞紧密连接的作用以及对Occludin表达的影响.方法 SD大鼠24只,按随机数字表法分为假手术组、失血性休克0.5h组、2h组、4h组,每组6只.采用Wiggers改良法制备失血性休克大鼠模型.腹主动脉取血,活性比色法测定血浆二胺氧化酶(diamine oxidase,DAO)活性;分光光度计法测定血浆D-乳酸(D-Lactate,D-LA)含量;比浊法测定血浆内毒素含量;肠系膜淋巴结和肝组织细菌培养检测细菌移位情况;光镜观察大鼠肠黏膜病理变化;电镜观察肠上皮细胞超微结构;RT-PCR测定小肠黏膜Occludin的mRNA表达;Westem blot与免疫组化法测定Occludin蛋白表达. 结果失血性休克0.5h组DAO、D-LA及内毒素含量分别为(72.68 ±10.24) mg/ml、(9.28±0.53) μg/ml、(41.25±3.68) pg/L,较假手术组明显升高(P＜0.05).2h组肝组织和肠系膜淋巴结均培养出细菌,4h组菌落量明显增高(P＜0.01).小肠上皮的形态及超微结构在休克0.5h组即发生破坏,随着失血时间延长,破坏越严重.失血性休克0.5h组肠黏膜OccludinmRNA降低(P＜0.01);2 h组降低更明显(P＜0.05).失血性休克0.5h组Occludin蛋白定位表达强度呈弱阳性;2h组Occludin蛋白量较0.5h组明显降低(P＜0.0l),定位表达亦呈阴性. 结论 Occludin分子在大鼠肠黏膜上皮细胞中的表达随失血性休克的时间延长下降,推测可能是导致细胞紧密连接损伤的重要基础.
목적 탐색실혈성휴극대장점막세포긴밀련접적작용이급대Occludin표체적영향.방법 SD대서24지,안수궤수자표법분위가수술조、실혈성휴극0.5h조、2h조、4h조,매조6지.채용Wiggers개량법제비실혈성휴극대서모형.복주동맥취혈,활성비색법측정혈장이알양화매(diamine oxidase,DAO)활성;분광광도계법측정혈장D-유산(D-Lactate,D-LA)함량;비탁법측정혈장내독소함량;장계막림파결화간조직세균배양검측세균이위정황;광경관찰대서장점막병리변화;전경관찰장상피세포초미결구;RT-PCR측정소장점막Occludin적mRNA표체;Westem blot여면역조화법측정Occludin단백표체. 결과 실혈성휴극0.5h조DAO、D-LA급내독소함량분별위(72.68 ±10.24) mg/ml、(9.28±0.53) μg/ml、(41.25±3.68) pg/L,교가수술조명현승고(P＜0.05).2h조간조직화장계막림파결균배양출세균,4h조균락량명현증고(P＜0.01).소장상피적형태급초미결구재휴극0.5h조즉발생파배,수착실혈시간연장,파배월엄중.실혈성휴극0.5h조장점막OccludinmRNA강저(P＜0.01);2 h조강저경명현(P＜0.05).실혈성휴극0.5h조Occludin단백정위표체강도정약양성;2h조Occludin단백량교0.5h조명현강저(P＜0.0l),정위표체역정음성. 결론 Occludin분자재대서장점막상피세포중적표체수실혈성휴극적시간연장하강,추측가능시도치세포긴밀련접손상적중요기출.
Objective To investigate the effect of hemorrhagic shock on intestinal epithelial tight junction and expression of protein Occludin.Methods Twenty-four SD rats were allocated to sham operation group,hemorrhagic shock 0.5-hour,2-hour,and 4-hour groups according to random number table,with 6 rats per group.A model of hemorrhagic shock in the rat was induced by the improved Wiggers method.Blood samples were drawn from abdominal aorta.Activity of plasma diamine oxidase (DAO) was determined by active colonmetric method; content of plasma D-lactic acid (D-LA) by spectrophotometer method; plasma endotoxin levels by turbidimetry; bacterial translocation by bacterial culture of mesenteric lymph nodes and liver tissues; pathological changes of intestinal mucosa by light microscope; ultrastructure of intestinal epithelial cells by electron microscopy; Occludin mRNA expression in small intestinal mucosa by RT-PCR; Occludin expression by Western blot and imnmnohistochemical method.Results Levels of DAO [(72.68-± 10.24) mg/ml],D-LA [(9.28 ± 0.53) μg/ml] and endotoxin [(41.25 ±3.68) pg/L] in hemorrhagic shock 0.5-hour group increased markedly as compared with those in sham operation group (P ＜0.05).Bacteria were cultivated from the liver tissues and mesenteric lymph nodes in hemorrhagic shock 2-hour group and much more colonies of bacteria were observed in hemorrhagic shock 4-hour group (P ＜ 0.01).Morphology and ultrastructure of the small intestinal epithelium manifested immediate damage in hemorrhagic shock 0.5-hour group and much worse detriment was obsevved with a prolonged bleeding time.Occludin mRNA expression in small intestinal mucosa decreased in hemorrhagic shock 0.5-hour group,followed by a further reduction in hemorrhagic shock 2-hour group (P ＜0.05).Localized expression of Occludin in hemorrhagic shock O.5-hour group was weakly positive.Occludin content in hemorrhagic shock 2-hour group showed significant reduction when compared with that in hemorrhagic shock O.5-hour group (P ＜ 0.01) and its localized express was negative as well.Conclusion Occludin expression in intestinal epithelial cells decreases obviously with a prolonged bleeding time,which is presumed to be an important basis for disruption of cellular tight junction.