乌司他丁对脓毒血症大鼠肠黏膜屏障损伤的保护及其对Wnt信号通路的影响

doi:10.16098/j.issn.0529-1356.2021.02.021

摘要/Abstract

摘要：

目的探讨乌司他丁（尿抑制素，UTI）能否对大鼠脓毒症肠黏膜损伤提供保护及其机制。方法选取SD大鼠100只，采用计算机软件随机分为对照组、脓毒症组、乌司他丁组、XAV939+乌司他丁组、氧化锂（LiCl ）+乌司他丁组。以经典盲肠结扎穿孔法建立脓毒症模型，检查评估空肠黏膜的损伤。采用酶联免疫吸附法(ELISA)检测炎症因子白细胞介素（IL-6）及肿瘤坏死因子（TNF-α）水平，采用Real-time PCR、Western blotting检测乌司他丁对β-连环蛋白（β-catenin）和细胞周期蛋白（cyclin D1）的表达情况；观察XAV939阻断或者LiCl激活Wnt信号通路对乌司他丁保护大鼠肠黏膜及Wnt信号通路相关蛋白的影响。结果脓毒症组IL-6、TNF-α水平及肠道黏膜损伤评分均显著高于乌司他丁组；脓毒症组β-catenin及cyclin D1的mRNA和蛋白表达水平均较对照组显著升高，差异具有显著性（P<0.05），给予乌司他丁处理之后，β-catenin及cyclin D1 mRNA和蛋白表达水平均显著降低，差异具有显著性（P<0.05）；与乌司他丁相比，XAV939促进了乌司他丁对大鼠肠黏膜的保护的作用，而且β-catenin和cyclin D1的蛋白表达降低（P<0.05）；LiCl减弱了乌司他丁对大鼠肠黏膜的保护作用，而且β-catenin和cyclin D1的蛋白表达升高，差异具有显著性（P<0.05）。结论乌司他丁通过下调β-catenin的表达抑制Wnt信号通路，降低炎症因子IL-6、TNF-α表达，从而改善脓毒症导致的肠黏膜屏障功能损伤。

关键词: 乌司他丁, 脓毒症, 肠黏膜, Wnt信号通路, 免疫印迹法, 大鼠

Abstract:

Objective To investigate the intestinal mucosal barrier function protective effect of ulinastatin in sepsis rats and its effect on Wnt/β-catenin signaling pathway. Methods One hundred SD rats were randomly divided into control group, sepsis group, ulinastatin group, XAV939+ulinastatin group and lithium chloride(LiCl)+ulinastatin group. The classical cecal ligation was used to duplicate sepsis model, and the jejunal mucosal injury was evaluated. The levels of inflammatory factors interleukin(IL)-6 and tumor necrosis factor(TNF)-α were detected by ELISA, and the expressions of β-catenin and cyclin D1 were detected by Real-time PCR and Western blotting. We also observed the effect of the Wnt signal pathway blockage by XAV939 or Wnt signal pathway activator by LiCl on ulinastatin protection of intestinal mucosa and proteins related to the Wnt signal pathway. Results The levels of IL-6, TNF-α and intestinal mucosal injury in the sepsis group were significantly higher than those in the ulinastatin group. The mRNA and protein expression levels of β-catenin and cyclin D1 in the sepsis group were significantly higher than those in the control group (P<0.05), After ulinastatin treatment, the expression levels of β-catenin and cyclin D1 mRNA and protein were significantly decreased, and the difference was significant (P<0.05). Compared with the ulinastatin group, combined treatment with XAV939 promoted the protective effect of ulinastatin on the intestinal mucosa of rats, and the protein expression of β-catenin and cyclin D1 was reduced (P<0.05). Combined treatment with LiCl weakened the protective effect of ulinastatin on the intestinal mucosa of rats, and the protein expression of β-catenin and cyclin D1 was increased (P<0.05). Conclusion Ulinastatin may inhibit the Wnt signaling pathway by down-regulating the expression of β-catenin, reduce the expression of inflammatory factors IL-6 and TNF-α, thereby promote repairing the intestinal mucosal barrier function damage.

Key words: Ulinastatin, Sepsis, Intestinal mucosa, Wnt signal pathway, Western blotting, Rat

中图分类号:

R459.7

王宇辉叶八宁吴婧龙大利李琨施贤清. 乌司他丁对脓毒血症大鼠肠黏膜屏障损伤的保护及其对Wnt信号通路的影响[J]. 解剖学报, 2021, 52(2): 295-299.

WANG Yu-hui YE Ba-ning WU Jing LONG Da-li LI Kun SHI Xian-qing. Effects of ulinastatin on intestinal mucosal barrier function in sepsis rats and its effect on wnt signal transduction pathway[J]. Acta Anatomica Sinica, 2021, 52(2): 295-299.

参考文献

［1］ Jin PF, Chen L, Hu YR, et al. Progress of study on intestinal pathogenesis of sepsis［J］. Chinese Journal of Nosocomiology,2018,28(10):1441-1445. (in Chinese)

金鹏锋,陈琳,胡耀仁,等.脓毒症的肠道发病机制研究进展［J］.中华医院感染学杂志,2018,28(10):1441-1445.

［2］ Khosravi A, Mazmanian SK. Disruption of the gut microbiome as a risk factor for microbial infections［J］. Curr Opin Microbiol,2013,16(2):221-227.

［3］ Shi LL, Han YQ, Ren HJ, et al. Research advance of pathology and physiology of sepsis［J］.Chinese Journal of Nosocomiology,2016,26(8):1914-1916. (in Chinese)

师灵灵,韩艳秋,任慧娟,等.脓毒症的病理生理机制研究进展［J］.中华医院感染学杂志,2016,26(8):1914-1916.

［4］ Yang J, Radulescu A, Chen CL, et al. Heparin-binding epidermal growth factor-like growth factor improves intestinal barrier function and reduces mortality in a murine model of peritonitis［J］. Surgery,2013,153(1):52-62.

［5］ Linder A, Russell JA. Russell an exciting candidate therapy for sepsis: ulinastatin, a urinary protease inhibitor［J］. Intensive Care Med, 2014,40(8):1164-1167.

［6］ Han D, Shang W, Wang G, et al. Ulinastatin- and thymosin α1-based immunomodulatory strategy for sepsis: a meta-analysis［J］. Int Immunopharmacol,2015, 29(2):377-382.

［7］ Rodríguez-González R, Martín-Barrasa JL, Ramos-Nuez á, et al. Multiple system organ response induced by hyperoxia in a clinically relevant animal model of sepsis［J］. Shock, 2014, 42(2):148-153.

［8］ Pugia MJ, Lott JA. Pathophysiology and diagonostic value of urinary trypsin inhibitors［J］. Clin Chem Lab Med, 2005,43(1):1-16.

［9］ Chen ZhP. Effect of wnt/β-catenin signaling pathway on acute lung injury in sepsis［D］. Wuhan: Huazhong University of Science and Technology,2016. (in Chinese)

陈正平. Wnt/β-catenin信号通路对脓毒症急性肺损伤的影响［D］.武汉：华中科技大学,2016.

［10］ Cai,YY, Zhou JP, Hu Q, et al. Effect of Wnt/β-catenin signaling pathway on the repair of orthodontic root, resorption［J］. Journal of Chongqing Medical University,2018,43(8):1109-1115. (in Chinese)

蔡洋伊,周建萍,胡琴,等.Wnt/β-catenin信号通路对正畸牙根吸收后修复作用的研究［J］.重庆医科大学学报,2018,43(8):1109-1115.

［11］ Wang H, Mao JL, Wu YM, et al. Protective effect of Zhenrenyangzang decoction on intestinal mucosal bar-rier function in ulcerative colitis rats［J］. Chinese Journal of Pathophysiology,2017,33(11):2053-2059. (in Chinese)

王慧,毛晶磊,吴艳敏,等.真人养脏汤对溃疡性结肠炎大鼠肠道黏膜屏障功能的保护作用［J］.中国病理生理杂志,2017,33(11):2053-2059.

［12］ Schmittgen TD, Livak KJ. Analyzing real-time PCR data by the comparative C(T) method［J］. Nat Protoc, 2008, 3(6): 1101-1108.

［13］ Xie F, Min S, Chen J, et al. Ulinastatin inhibited sepsis-induced spinal inflammation to alleviate peripheral neuromuscular dysfunction in an experimental rat model of neuromyopathy［J］. J Neurochem, 2017,143(2): 225-235.

［14］ Liang Q, Diao BB, Wang C. Effect of Yiqi Yangyin Huoxue recipe on serum TNF-α, IL-6 and IL-10 in septic model rats［J］. Chinese Journal of Immunology, 2014,11(12):1705-1707. (in Chinese)

梁群, 刁冰冰, 王丛. 益气养阴活血法对脓毒症模型大鼠血清TNF-α、IL-6与IL-10的影响［J］.中国免疫学杂志, 2014,11(12):1705-1707.

［15］ Tian A, Benchabane H, Wang Z, et al. Regulation of stem cell proliferation and cell fate specification by Wingless/Wnt signaling gradients enriched at adult intestinal compartment boundaries［J］. PLoS Genetics, 2016, 12(2):e1005822.

［16］ Yao LH. Advanced glycation end product receptors participate in the regulation of TDI asthma airway inflammation by stabilizing β-catenin signaling［D］. Guangzhou: Southern Medical University,2017. (in Chinese)

姚利红. 晚期糖基化终末产物受体通过稳定β-catenin信号参与调控TDI哮喘气道炎症［D］.广州：南方医科大学,2017.

［17］ Zhu YT, Guo Y, Qiao YY, et al. Verbenalin inhibits airway inflammation and remodeling by modulating Wnt/β-catenin signal pathway in asthma rats［J］. The Journal of Practical Medicine,2018,34(17):2880-2883. (in Chinese)

朱颖涛,郭燕,乔岩岩,等.马鞭草苷通过Wnt/β-catenin信号通路干预哮喘大鼠气道炎症及气道重塑的研究［J］.实用医学杂志,2018,34(17):2880-2883.

[1]	洪晓敏李三强崔钦奕郑润月杨孟利罗仁利李前辉 . 酒精性肝纤维化核因子κB信号通路与性别的差异 [J]. 解剖学报, 2024, 55(1): 55-61.
[2]	谢惠兰方芳林毅. Chir99021调控Wnt/β-catenin 信号通路抑制大鼠牙髓干细胞成骨诱导分化的机制[J]. 解剖学报, 2024, 55(1): 67-72.
[3]	魏茜茜李超红赵晨露唐家萍刘玉珍. 大鼠颈上神经节中Ⅰ组代谢型谷氨酸受体表达及慢性间歇性低氧对其的影响 [J]. 解剖学报, 2024, 55(1): 3-9.
[4]	袁蕾杨智伟杨惠刘政海何洁万炜. 三七总皂苷抑制小鼠星形胶质细胞活化缓解完全弗氏佐剂所致炎性痛 [J]. 解剖学报, 2024, 55(1): 25-31.
[5]	马婷婷陈建红刘爱翠李海宁. 抑制lncRNA TUG1下调核苷酸结合寡聚结构域样受体蛋白1炎症小体在延缓阿尔茨海默病进展的作用 [J]. 解剖学报, 2024, 55(1): 32-42.
[6]	邹成功冯浩陈兵唐辉邵川孙谋杨荣何家全. 创伤性颅脑损伤中神经功能障碍与认知功能损伤的动态变化 [J]. 解剖学报, 2024, 55(1): 43-48.
[7]	吕海燕沈西雅赵富生朱梅. 松茸多糖对 1-甲基-4-苯基-吡啶离子诱导 PC12 细胞损伤的影响 [J]. 解剖学报, 2024, 55(1): 49-54.
[8]	郑丽平刘霞杜晓华吴亚娟刘珊珊 . 脑源性神经营养因子在牦牛与黄牛端脑不同区域的表达与分布 [J]. 解剖学报, 2024, 55(1): 10-16.
[9]	苏芃王兴仪梁景岩熊天庆. 一种低密度及高纯度胎鼠原代海马神经元培养方法的优化及鉴定[J]. 解剖学报, 2024, 55(1): 113-119.
[10]	郝永明郭磊周程辉裴汉军李莉赵哲 . 改良腹主动脉缩窄法建立心肌肥厚模型[J]. 解剖学报, 2024, 55(1): 120-124.
[11]	许亚平余悦欣陈金福王柯柯郭志坤 . 高龄大鼠心室肌间质弹性纤维和胶原纤维的结构分布特征及其机制 [J]. 解剖学报, 2023, 54(6): 716-721.
[12]	刘丽平朱梦妮徐慧 . 白细胞介素6对脂多糖诱导所致类子痫前期大鼠有核红细胞的影响 [J]. 解剖学报, 2023, 54(6): 722-729.
[13]	赵卫明李晓余国营王改平常翠芳 . 丝氨酸肽酶抑制因子Kazal型1对肝癌细胞增殖的影响及其机制 [J]. 解剖学报, 2023, 54(6): 695-702.
[14]	王文娟丁雨桐苏昊任捷孙艳荣王涵菲陆嘉莉张林倩白钰秦丽华. 低雌激素状态下大鼠海马及纹状体Nogo-A的表达变化[J]. 解剖学报, 2023, 54(6): 620-627.
[15]	张琴杨俊霞蒋青松. 福辛普利对糖尿病视网膜病变小鼠血管紧张素转化酶2和血管内皮生长因子的影响[J]. 解剖学报, 2023, 54(6): 628-634.