胆汁内外引流术对梗阻性黄疸大鼠血浆二胺氧化酶活性及肠源性防御素-5mRNA表达水平的影响

发布时间：2018-06-27 08:56

本文选题：梗阻性黄疸 + 胆汁引流　；参考：《中国人民解放军医学院》2014年硕士论文

【摘要】：【研究目的】1、观察梗阻性黄疸（梗黄）形成与解除大鼠肠黏膜组织形态、血浆二胺氧化酶（Diamine Oxidase,DAO）的活性变化，探讨二者之间的关系，为临床建立敏感无创的检测方法来判断肠黏膜屏障功能的损伤和修复状况提供实验及理论依据；2、观察大鼠防御素-5（Rat Defensin-5,RD-5）mRNA的变化，探讨胆汁内引流术优于外引流术的机制，为临床治疗方式的选择提供实验依据。【材料与方法】将60只成年雄性SD大鼠随机分为四组：梗阻性黄疸组(obstructive jaundice,OJ)、胆汁外引流组(external drainage,ED)、胆汁内引流组(internal drainage,ID)及假手术对照组(shame operation,SH)，各组均15只。建立动物模型：第一次手术建立SH模型（只分离胆管不结扎）和OJ模型（分离胆管并结扎、切断），，OJ术后7天行二次手术，建立ED模型（胆管插管经皮引流至体外）和ID模型（胆管与十二指肠间建立引流）。造模成功后，处死大鼠并采集标本，下腔静脉取血ELASA方法检测各组大鼠血浆DAO的活性;取末段回肠病理切片、HE染色,光镜下观察末端回肠组织黏膜形态学变化；取末段回肠RT-PCR方法检测RD-5mRNA的表达水平。【结果】1.胆管梗阻形成后，OJ组大鼠肠黏膜损伤明显，黏膜变薄、绒毛稀疏、上皮完整性受到破坏并伴有炎性细胞浸润，Chiu氏评分为2.158±0.579，ID、ED组大鼠肠黏膜有不同程度的恢复（Chiu氏评分分别为0.633±0.144，1.329±0.289），ID组更接近SH组（0.315±0.080），四组间两两比较P值均0.01；OJ组血浆DAO水平显著升高，明显高于SH、ID、ED组（8.183±0.211vs3.570±0.280、3.978±0.269、5.051±0.328U/L，P0.01），引流术后DAO水平下降，ID组较ED组下降更明显，二者差异有统计学意义（P0.01），ID组血浆DAO水平略高于SH组，二者差异无统计学意义（P0.05）；各大鼠血浆DAO水平与肠粘膜损伤评分的相关系数r=0.89(P0.01)，大鼠血浆DAO变化与肠黏膜组织病理学变化一致，呈正相关。2.OJ组回肠RD-5mRNA的表达水平显著降低，明显低于SH、ID、ED组(0.0708±0.0061vs0.1895±0.0124、0.1903±0.0143、0.1601±0.0078，P0.01），引流术后RD-5mRNA的表达水平升高，ID组较ED组升高更明显，二者差异有统计学意义（P0.01），ID组RD-5mRNA表达水平略高于SH组，二者差异无统计学意义（P0.05）。【结论】梗黄后，机体高胆红素血症及肠道胆汁缺乏致肠黏膜上皮受损，血浆DAO活性升高，胆汁内外引流术可修复肠黏膜屏障，逆转血浆DAO水平，RD-5mRNA表达水平增高，且以内引流效果更佳。血浆DAO水平变化与肠黏膜形态学改变具有一致性，可反映梗黄大鼠肠黏膜屏障损伤及修复情况；胆汁内引流优于外引流的机制之一可能在于肠源性防御素表达水平更高，抗菌作用更强、更能促进肠道黏膜免疫的恢复。
[Abstract]:[Objective] 1 to observe the formation of obstructive jaundice (stem yellow) and the changes in the tissue morphology of intestinal mucosa and the changes of the activity of plasma two amine oxidase (Diamine Oxidase, DAO) in rats, and explore the relationship between the two, and provide an experimental and theoretical basis for the establishment of a sensitive and noninvasive detection method to judge the damage and repair of the intestinal mucous membrane barrier function. 2, 2, observe the changes of Rat Defensin-5 (RD-5) mRNA of rat, and explore the mechanism of biliary drainage better than external drainage, and provide experimental basis for the choice of clinical treatment.
[materials and methods] 60 adult male SD rats were randomly divided into four groups: the obstructive jaundice group (obstructive jaundice, OJ), the external biliary drainage group (external drainage, ED), the bile drainage group (internal drainage, ID) and the sham operation control group (shame operation,), all of the groups were 15. Type (isolated bile duct without ligation) and OJ model (separating bile duct and ligating and cutting off), two operations were performed 7 days after OJ. The ED model (bile duct intubation percutaneous drainage to external drainage) and ID model (drainage between bile duct and duodenum) were established. After the success of the model, the rats were killed and the specimens were collected, and the blood plasma DAO of the inferior vena cava was taken ELASA method to detect the plasma DAO in each group. The pathological sections of the last ileum, HE staining, and light microscopy were used to observe the morphological changes of the mucosa of the distal ileum tissue, and the expression level of RD-5mRNA was detected by the distal ileum RT-PCR method.
[results] after the formation of 1. bile duct obstruction, the intestinal mucosa of OJ rats was significantly damaged, the mucosa thinned, the villus was sparse, the epithelial integrity was damaged and inflammatory cell infiltration, the Chiu score was 2.158 + 0.579, ID, the intestinal mucosa of the group ED rats had different degrees of recovery (Chiu's score was 0.633 + 0.144,1.329 + 0.289 respectively), and the ID group was closer to the SH group (0.3 15 + 0.080), 22 of the four groups, the P value was 0.01, and the level of plasma DAO in group OJ was significantly higher than that in SH, ID, ED group (8.183 + 0.211vs3.570 + 0.280,3.978 + 0.269,5.051 + 0.328U/L, P0.01). The decrease of DAO level after drainage was more obvious than that in the group of two. The level of two groups was slightly higher than that of the group, two There was no statistical significance (P0.05). The correlation coefficient of plasma DAO level and intestinal mucosal injury score of rats was r=0.89 (P0.01), and the changes of DAO in plasma of rats were in accordance with the pathological changes of intestinal mucosa, and the expression level of the ileum RD-5mRNA in the positive correlation group was significantly lower than that of the SH, ID, ED group (0.0708 + 0.0061vs0.1895 + 0.0124,0.1903). 0.0143,0.1601 + 0.0078, P0.01), the expression level of RD-5mRNA increased after drainage, and the increase in ID group was more obvious than that in ED group. The difference between the two groups was statistically significant (P0.01), and the level of RD-5mRNA in the ID group was slightly higher than that in the SH group, and the difference between the two groups was not statistically significant (P0.05).
[Conclusion] the intestinal mucosal epithelium was damaged by hyperbilirubinemia and intestinal bile deficiency, the activity of plasma DAO increased, the internal and external drainage of bile can repair the intestinal mucosal barrier, reverse the plasma DAO level, increase the level of RD-5mRNA expression, and the effect of internal drainage is better. The change of plasma DAO level is consistent with the morphological changes of intestinal mucosa. It can reflect the damage and repair of intestinal mucosal barrier in the stem yellow rats. One of the mechanisms of bile drainage is better than the external drainage mechanism, which may lie in the higher expression level of enteric defensin, stronger antibacterial effect, and can promote the recovery of intestinal mucosal immunity.
【学位授予单位】：中国人民解放军医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R575

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