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中国临床药理学与治疗学 ›› 2022, Vol. 27 ›› Issue (5): 544-550.doi: 10.12092/j.issn.1009-2501.2022.05.007

• 基础研究 • 上一篇    下一篇

Rho激酶抑制剂对脓毒症肝损伤的影响

邢博民1,郭娜1,宁海慧1,丁新耘1,马玉清2   

  1. 1兰州大学第一临床医学院,兰州 730000,甘肃;2兰州大学第一医院麻醉科,兰州 730000,甘肃
  • 收稿日期:2021-12-22 修回日期:2022-04-14 出版日期:2022-05-26 发布日期:2022-06-06
  • 通讯作者: 马玉清,女,医学博士,主任医师,教授,硕士生导师,研究方向:脓毒症。 E-mail: myq2392466@163.com
  • 作者简介:邢博民,男,硕士研究生,研究方向:脓毒症脏器保护。 E-mail: m13255508762@163.com
  • 基金资助:
    甘肃省自然科学基金(21JR1RA077)

Effects of Rho kinase inhibitors on septic liver injury

XING Bomin1, GUO Na1, NING Haihui1, DING Xinyun1, MA Yuqing2   

  1. 1The First Clinical Medical College of Lanzhou University, Lanzhou 730000, Gansu, China; 2Department of Anesthesiology, the First Hospital of Lanzhou University, Lanzhou 730000, Gansu, China
  • Received:2021-12-22 Revised:2022-04-14 Online:2022-05-26 Published:2022-06-06

摘要: 目的:探讨Rho激酶抑制剂Y-27632对脓毒症急性肝损伤的作用。方法:健康成年雄性SD大鼠32只,随机分为假手术组(Sham组)、假手术+Y-27632组(Sham+Y组),盲肠结扎穿孔组(CLP组)和CLP+Y-27632组(CLP+Y组),每组8只。采用CLP方法制备大鼠脓毒症模型,造模24 h心脏采血后安乐死大鼠,收集血清及肝脏组织。苏木精-伊红(HE)染色观察各组大鼠肝组织病理学变化;蛋白质免疫印迹(Western blot)检测各组大鼠肝组织ROCK1和下游NF-κB蛋白的表达情况;免疫组织化学法(ICH)检测大鼠肝脏组织ROCK1蛋白表达;酶联免疫吸附(ELISA)法检测各组大鼠血清肝功能指标ALT、AST水平及肝组织匀浆IL-18、IL-10和谷胱甘肽(GSH)含量变化。结果:与Sham组相比,Sham+Y组肝脏组织病理学无明显变化,CLP组肝细胞排列紊乱,有大量炎症细胞浸润,CLP+Y组炎症细胞浸润较少,肝细胞条索逐渐恢复。与Sham组相比,CLP和CLP+Y组ROCK1蛋白表达增加(P<0.05);与CLP组相比,CLP+Y组ROCK1蛋白表达减少(P<0.05)。与Sham组相比,CLP和CLP+Y组NF-κB蛋白表达增加(P<0.05);与CLP组相比,CLP+Y组NF-κB蛋白表达减少(P<0.05);ICH检测到Sham组肝脏ROCK1少量表达,CLP组和CLP+Y组大量表达;与Sham组相比,CLP和CLP+Y组血清ALT、AST水平升高(P<0.05),与CLP组相比,CLP+Y组ALT、AST水平降低(P<0.05)。与Sham组相比,CLP和CLP+Y组肝组织匀浆IL-18含量升高(P<0.05),CLP组肝组织匀浆IL-10和GSH含量降低(P<0.05),CLP+Y组IL-10和GSH的变化相近(P>0.05);与CLP组相比,CLP+Y组IL-18含量降低(P<0.05),IL-10和GSH含量升高(P<0.05)。 结论:Rho激酶抑制剂可以减轻脓毒症大鼠急性肝损伤,可能与抑制Rho/ROCK/NF-κB信号通路及其下游的炎症因子,减轻肝脏炎症反应,同时降低肝脏氧化应激水平有关。

关键词: 脓毒症, 肝损伤, ROCK1蛋白, Y-27632

Abstract:

AIM: To investigate the effect of Rho kinase inhibitor Y-27632 on acute liver injury in sepsis. METHODS: Thirty-two healthy adult male SD rats were randomly divided into sham operation group (Sham group), sham operation+Y-27632 group (Sham+Y group), cecal ligation and perforation group (CLP group) and CLP+Y-27632 group (CLP+Y-27632 group), 8 animals in each group. The rat sepsis model was established by the CLP method, and the rat was euthanized 24 hours after the model was established, and the serum and liver tissue were collected. Hematoxylin-eosin (HE) staining was used to observe the pathological changes in the liver tissue of the rats in each group; Western Blot was used to detect the expressions of ROCK1 and downstream NF-κB proteins in the liver tissue of the rats in each group; immunohistochemical method The expression of ROCK1 protein in liver tissue of rats was detected; enzyme-linked immunosorbent assay (ELISA) method was used to detect the levels of serum liver function indexes ALT and AST, and the changes of IL-18, IL-10 and GSH contents in liver tissue homogenate. RESULTS: Compared with the Sham group, there was no significant change in the histopathology of the liver in the Sham+Y group. In the CLP group, the arrangement of hepatocytes was disordered, with a large number of inflammatory cells infiltrating. Compared with the Sham group, the expression of ROCK1 protein in the CLP and CLP+Y groups was increased (P<0.05); compared with the CLP group, the ROCK1 protein expression in the CLP+Y group was decreased (P<0.05). Compared with the Sham group, the expression of NF-κB protein in the CLP and CLP+Y groups increased (P<0.05); compared with the CLP group, the NF-κB protein expression in the CLP+Y group decreased (P<0.05); A small amount of expression was found in the liver Sham group, and a large amount was expressed in the CLP group and CLP+Y group; compared with the Sham group, the serum ALT and AST levels in the CLP and CLP+Y groups were increased (P<0.05). The levels of ALT and AST in +Y group decreased (P<0.05). Compared with the Sham group, the contents of IL-18 in the liver tissue homogenate of the CLP and CLP+Y groups increased (P<0.05), while the contents of IL-10 and GSH in the liver tissue homogenate of the CLP group decreased (P<0.05). The changes of IL-10 and GSH in the group were similar (P>0.05); compared with the CLP group, the content of IL-18 in the CLP+Y group was decreased (P<0.05), and the content of IL-10 and GSH was increased (P<0.05). CONCLUSION: Rho kinase inhibitor can alleviate acute liver injury in septic rats, which may be related to inhibiting the expression of ROCK1 and NF-κB proteins, reducing the inflammatory response of liver tissue, and reducing the level of liver oxidative stress.

Key words: sepsis, liver injury, ROCK1 protein, Y-27632

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