肾素血管紧张素系统在溃疡性结肠炎小鼠肠-血屏障损伤中的调控作用

doi:10.11843/j.issn.0366-6964.2024.04.037

摘要/Abstract

摘要： 本研究旨在探讨肾素血管紧张素系统(renin-angiotensin system, RAS)在溃疡性结肠炎小鼠肠-血屏障损伤中发挥的作用。选取16只ICR小鼠，随机分为正常对照组(Control组)和葡聚糖硫酸钠盐处理组(DSS组)，等体积灌胃生理盐水。试验至第4天，DSS组小鼠在饮用水中添加DSS(终浓度为4%)制造小鼠溃疡性结肠炎模型。期间每天记录小鼠体重、粪便性状和粪便隐血情况，并计算疾病活跃指数(disease activity index, DAI)。DSS饮用至第8天，所有小鼠采血后剖检，量取结肠长度，取结肠组织等样品。进行如下试验：1)HE染色观察结肠组织病理变化；2)ELISA法检测血液中血管内皮生长因子(vascular endothelial growth factor, VEGFA)及结肠组织中Ang1-7和Ang Ⅱ的含量；3)Western blot检测结肠组织中血管紧张素转化酶2(angiotensin-coverting enzyme 2, ACE2)、血管紧张素转化酶(ACE)和质膜膜泡关联蛋白(plasmalemma vesicle associated protein, PLVAP)的表达；4)斯皮尔曼相关性分析ACE2、ACE与PLVAP表达变化的相关性以及Ang1-7、Ang Ⅱ和VEGFA含量变化的相关性。结果显示：1)成功建立了DSS诱导小鼠溃疡性结肠炎模型，小鼠表现为体重下降、DAI极显著升高(P < 0.01)、结肠极显著缩短(P < 0.01)和结肠组织出现明显的病理变化；2)与对照组相比，DSS组小鼠表现肛门明显出血，结肠组织中PLVAP和VEGFA蛋白表达均极显著升高(P < 0.01)；3)与对照组相比，DSS组小鼠结肠组织中ACE2和ACE表达均极显著上调(P < 0.01)，Ang1-7和Ang Ⅱ含量分别显著(P < 0.05)和极显著(P < 0.01)升高；4)相关性分析显示，ACE2、ACE的表达变化和PLVAP表达变化呈正相关，Ang1-7、Ang Ⅱ含量变化与VEGFA含量变化也呈正相关。以上结果表明，DSS致小鼠结肠炎症过程中小鼠结肠血管屏障受损，结肠局部RAS均处于激活状态，ACE/Ang Ⅱ通路的激活占优势，提示：Ang Ⅱ参与了结肠炎小鼠肠-血屏障的损伤过程。通过激活ACE2或过表达ACE2，增强其对Ang Ⅱ的降解作用以缓解肠-血屏障的损伤，可能是治疗或缓解溃疡性结肠炎的一个新的思路或途径。

关键词: 肾素-血管紧张素系统, 小鼠, 溃疡性结肠炎, 肠血屏障

Abstract: The purpose of this study was to investigate the role of renin-angiotensin system (RAS) in gut-vascular barrier injury in ulcerative colitis mice. Sixteen ICR mice were selected and randomly divided into control and DSS groups by equal volume saline gavage. Mice in the DSS group were supplemented with DSS (final concentration of 4%) in the drinking water starting on day 4 of the study to establish a murine ulcerative colitis model. During this period, body weight, fecal characteristics, and fecal occult blood of the mice were recorded daily for the calculation of DAI. Until day 8 of DSS drinking, all mice were necropsied after blood collection, then the colon length was measured, and colon tissue samples were collected. The following experiments were performed: 1) HE staining to observe the histopathological changes in the colon of each group of mice; 2) ELISA to detect the levels of VEGFA in blood and Ang1-7 and Ang Ⅱ in colon tissues; 3) Western blot to detect the expression of ACE2, ACE and PLVAP in colon tissues; 4) Spearman correlation analysis was used to analyze the correlation between ACE2, ACE expression changes and PLVAP expression changes, as well as the correlation between Ang1-7 and Ang Ⅱ content changes and VEGFA content changes. Results: 1) A DSS-induced ulcerative colitis model was successfully established in mice, which showed decreased body weight, highly significant increase in DAI (P < 0.01), highly significant shortening of colon (P < 0.01) and severe pathological changes in colon tissues; 2) Compared with the control group, mice in the DSS group showed colonic hemorrhage, highly significant increase in PLVAP and VEGFA protein expression in colon tissues (P < 0. 01); 3) Compared with the control group, ACE2 and ACE expression in colon tissues of mice in the DSS group were highly significantly upregulated (P < 0.01), and the contents of Ang1-7 and Ang Ⅱ were significantly and significantly increased (P < 0.05 and P < 0.01); 4) Correlation analysis showed that there were positive correlations between ACE2 and ACE expression changes and PLVAP expression changes, as well as Ang1-7 and Ang Ⅱ content changes and VEGFA content changes. The above results suggest that the colonic vascular barrier is damaged during DSS-induced colonic inflammation, the two pathways of RAS in the colon are activated or out of balance, and the activation of ACE/Ang Ⅱ pathways is dominant, suggesting that ACE/Ang Ⅱ is involved in the injury of intestinal-blood barrier in mice with colitis. Activating ACE2 or overexpressing ACE2 to enhance its degradation of Ang Ⅱ to alleviate the damage of intestinal blood barrier may be a new idea or way to treat or alleviate ulcerative colitis.

Key words: renin-angiotensin system, mice, ulcerative colitis, gut-vascular barrier

中图分类号:

S852.21

张崇昊, 马畅, 李志强, 伍钢, 张源淑. 肾素血管紧张素系统在溃疡性结肠炎小鼠肠-血屏障损伤中的调控作用[J]. 畜牧兽医学报, 2024, 55(4): 1756-1765.

ZHANG Chonghao, MA Chang, LI Zhiqiang, WU Gang, ZHANG Yuanshu. The Role and Relationship of Renin-angiotensin System in Gut-vascular Barrier Injury in Ulcerative Colitis Mice[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(4): 1756-1765.

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