乌司他丁对SAP大鼠肠粘膜HMGB1表达及肠粘膜屏障变化的研究
发布时间：2018-11-08 06:47
【摘要】：目的：探讨乌司他丁对于重症急性胰腺炎（severe acute pancreatitis，SAP）大鼠肠粘膜高迁移率族蛋白B1（high mobility group box-1protein，HMGB1）表达及对肠粘膜屏障的作用，旨在为SAP的临床治疗提供实验依据。 方法：实验动物选用雌性斯普拉-道来（Sprague dawley,SD）大鼠54只，清洁级，体重220-250g，平均体重237.45g。将实验大鼠称重后标号，随机分为对照组、SAP组及乌司他丁干预组，每组18只，实验前全部大鼠禁食12小时，但不禁饮水。运用穿刺十二指肠并逆行胆胰管注射牛磺胆酸钠法[1]诱导建立大鼠SAP模型，对照组开腹并翻动胰腺组织两三下后，逐层缝合切口。干预组于造模成功后5分钟内，行尾缘静脉注射乌司他丁（1000IU/100g体质量），10分钟内注射完毕，对照组及SAP组分别于造模成功后5分钟，行尾缘静脉注射生理盐水。每一组按造模成功后6小时、12小时、24小时分为3个时间亚组，每组6只，并按照上述时间点分3批处死大鼠，每批6只。使用生化分析仪检测各组血清淀粉酶含量，观察各组胰腺组织学变化，应用酶联免疫吸附方法（ELISA）测定各组血清TNF-α含量，取小肠组织后匀浆并应用ELISA法测定肠道HMGB1浓度，分光光度法测肠粘膜二胺氧化酶（diamineoxidase，DAO）含量，同时测定肠道组织湿干比。 结果：造模时可见胰腺组织肿胀明显，并有点状出血，提示造模成功[1]。SAP组18只大鼠死亡10只，乌司他丁干预组死亡6只，对照组死亡3只，死亡大鼠被剔除后重新造模以补充数据。6小时对照组、SAP组及干预组血清淀粉酶、血清TNF-α含量、肠道HMGB1含量、肠粘膜二胺氧化酶含量、肠道组织湿干比和胰腺组织病理评分两两比较，差异有统计学意义(p0.05)。12小时对照组、SAP组及干预组上述指标两两比较，差异有统计学意义(p0.05)。24小时血清TNF-αSAP组与干预组相比差异无统计学意义(p0.05),但两组与对照组相比差异有统计学意义(p0.05)。其余指标两两比较，差异有统计学意义(p0.05)。 结论：采用穿刺十二指肠逆行胆胰管注射牛磺胆酸钠法可成功制造出SAP动物模型。乌司他丁可减少致炎因子HMGB1在肠道组织的表达，减少二胺氧化酶在肠粘膜的丢失，降低肠道组织湿干比，保护肠道粘膜屏障功能。
[Abstract]:Objective: to investigate the effect of ulinastatin on the expression of intestinal mucosal high mobility group protein B1 (high mobility group box-1protein,HMGB1 and intestinal mucosal barrier in rats with severe acute pancreatitis (severe acute pancreatitis,SAP), in order to provide experimental evidence for the clinical treatment of SAP. Methods: Fifty-four female Sprague dawley,SD rats, weighing 220-250 g, were selected as experimental animals, with an average weight of 237.45 g. After weighing the experimental rats were randomly divided into control group, SAP group and ulinastatin intervention group, 18 rats in each group. All rats fasted for 12 hours before the experiment, but could not help drinking water. The rat SAP model was induced by injection of sodium taurocholate into the duodenum and retrograde cholangiopancreatic duct. In the control group, the abdominal cavity was opened and the pancreatic tissue was turned over for two or three times, and the incision was sutured layer by layer. In the intervention group, ulinastatin (1000IU/100g body mass) was injected intravenously in the caudal margin within 5 minutes after the successful modeling, and the control group and the SAP group were injected with normal saline in the caudal vein within 5 minutes after the successful establishment of the model. Each group was divided into 3 subgroups according to 6 hours, 12 hours and 24 hours after successful modeling, 6 rats in each group were killed in 3 batches according to the above time points, 6 rats were killed in each batch. The content of serum amylase in each group was detected by biochemical analyzer, and the histological changes of pancreas in each group were observed. The content of TNF- 伪 in serum of each group was determined by enzyme linked immunosorbent assay (ELISA), the homogenate of small intestine tissue was taken out and the concentration of HMGB1 in intestine was determined by ELISA method. The content of diamine oxidase (diamineoxidase,DAO) in intestinal mucosa and the wet / dry ratio of intestinal tissue were measured by spectrophotometry. Results: the pancreatic tissue swelling and punctate hemorrhage could be seen in the model making. [1] .Eighteen rats died in SAP group, 6 rats in ulinastatin intervention group and 3 rats in control group. Serum amylase, serum TNF- 伪, intestinal HMGB1 and intestinal mucosal diamine oxidase were found in 6 hour control group, SAP group and intervention group. The wet-to-dry ratio of intestinal tissue and the pathological score of pancreatic tissue were significantly different (p0.05). The above indexes were compared in 12 hours control group, SAP group and intervention group. There was no significant difference between the 24 hour serum TNF- 伪 SAP group and the intervention group (p0. 05), but there was a significant difference between the two groups and the control group (p0. 05). There was significant difference between the other indexes (p 0.05). Conclusion: SAP animal model can be successfully established by injection of sodium taurocholate into retrograde cholangiopancreatic duct of duodenum. Ulinastatin could reduce the expression of HMGB1 in intestinal tissue, decrease the loss of diamine oxidase in intestinal mucosa, decrease the wet / dry ratio of intestinal tissue, and protect intestinal mucosal barrier function.
【学位授予单位】：皖南医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R965
本文编号：2317693
[Abstract]:Objective: to investigate the effect of ulinastatin on the expression of intestinal mucosal high mobility group protein B1 (high mobility group box-1protein,HMGB1 and intestinal mucosal barrier in rats with severe acute pancreatitis (severe acute pancreatitis,SAP), in order to provide experimental evidence for the clinical treatment of SAP. Methods: Fifty-four female Sprague dawley,SD rats, weighing 220-250 g, were selected as experimental animals, with an average weight of 237.45 g. After weighing the experimental rats were randomly divided into control group, SAP group and ulinastatin intervention group, 18 rats in each group. All rats fasted for 12 hours before the experiment, but could not help drinking water. The rat SAP model was induced by injection of sodium taurocholate into the duodenum and retrograde cholangiopancreatic duct. In the control group, the abdominal cavity was opened and the pancreatic tissue was turned over for two or three times, and the incision was sutured layer by layer. In the intervention group, ulinastatin (1000IU/100g body mass) was injected intravenously in the caudal margin within 5 minutes after the successful modeling, and the control group and the SAP group were injected with normal saline in the caudal vein within 5 minutes after the successful establishment of the model. Each group was divided into 3 subgroups according to 6 hours, 12 hours and 24 hours after successful modeling, 6 rats in each group were killed in 3 batches according to the above time points, 6 rats were killed in each batch. The content of serum amylase in each group was detected by biochemical analyzer, and the histological changes of pancreas in each group were observed. The content of TNF- 伪 in serum of each group was determined by enzyme linked immunosorbent assay (ELISA), the homogenate of small intestine tissue was taken out and the concentration of HMGB1 in intestine was determined by ELISA method. The content of diamine oxidase (diamineoxidase,DAO) in intestinal mucosa and the wet / dry ratio of intestinal tissue were measured by spectrophotometry. Results: the pancreatic tissue swelling and punctate hemorrhage could be seen in the model making. [1] .Eighteen rats died in SAP group, 6 rats in ulinastatin intervention group and 3 rats in control group. Serum amylase, serum TNF- 伪, intestinal HMGB1 and intestinal mucosal diamine oxidase were found in 6 hour control group, SAP group and intervention group. The wet-to-dry ratio of intestinal tissue and the pathological score of pancreatic tissue were significantly different (p0.05). The above indexes were compared in 12 hours control group, SAP group and intervention group. There was no significant difference between the 24 hour serum TNF- 伪 SAP group and the intervention group (p0. 05), but there was a significant difference between the two groups and the control group (p0. 05). There was significant difference between the other indexes (p 0.05). Conclusion: SAP animal model can be successfully established by injection of sodium taurocholate into retrograde cholangiopancreatic duct of duodenum. Ulinastatin could reduce the expression of HMGB1 in intestinal tissue, decrease the loss of diamine oxidase in intestinal mucosa, decrease the wet / dry ratio of intestinal tissue, and protect intestinal mucosal barrier function.
【学位授予单位】：皖南医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R965
【参考文献】
相关期刊论文 前10条
1 黎君友，于燕，郝军，，晋桦，许惠君;分光光度法测定血和小肠组织二胺氧化酶活性[J];氨基酸和生物资源;1996年04期
2 唐晋;宋孟龙;程飞;张献全;;大承气汤治疗重症急性胰腺炎大鼠的实验研究[J];重庆医学;2010年09期
3 ;Low-dose dopamine reduces inflammatory factors of acute pancreatitis in rats[J];Hepatobiliary & Pancreatic Diseases International;2007年06期
4 刁春香,商弘,王世龙;善得定与洛塞克联合治疗重症胰腺炎[J];黑龙江医学;2002年07期
5 万美华;李娟;唐文富;龚翰林;陈光远;薛平;赵先林;夏庆;;大承气汤对急性胰腺炎致急性肺损伤-腹内高压的作用[J];四川大学学报(医学版);2011年05期
6 方立峰;;乌司他丁治疗急性坏死性胰腺炎的实验研究[J];局解手术学杂志;2008年04期
7 李君久,覃谦,王力斌,李洪;生长抑素治疗轻型急性胰腺炎的疗效研究[J];宁夏医学杂志;2004年03期
8 栾正刚;张成;葛春林;马晓春;郭仁宣;;重症急性胰腺炎大鼠HMGB1表达与肠黏膜屏障损害的关系[J];中国现代普通外科进展;2008年01期
9 林栋栋,孙家邦,李非,张淑文,崔叶青,刘大川,朱斌,孙海晨,刘爽;IL-8、IL-10在大鼠急性胰腺炎并发肺损伤中的作用[J];首都医科大学学报;2005年02期
10 张明钧;姚玮艳;乔敏敏;诸琦;袁耀宗;;肠壁穿刺逆行胰胆管注射牛黄胆酸钠重症急性胰腺炎造模[J];上海交通大学学报(医学版);2006年05期
本文编号：2317693
本文链接：https://www.wllwen.com/yixuelunwen/yiyaoxuelunwen/2317693.html
最近更新
教材专著
