异莲心碱通过抑制TLR4减轻脂多糖诱导的急性肺损伤作用机制的研究

职业与健康 ›› 2023, Vol. 39 ›› Issue (22): 3067-3071.

异莲心碱通过抑制TLR4减轻脂多糖诱导的急性肺损伤作用机制的研究

李文^a, 宋杨一嫣^b, 宋金春^a

武汉大学人民医院a急诊科,b药学部,湖北武汉 430060

收稿日期:2023-04-11 修回日期:2023-05-09 出版日期:2023-11-15 发布日期:2026-03-26
通信作者: 宋杨一嫣,副主任药师,E-mail：919572095@qq.com
作者简介:李文,男,副主任医师,主要从事心肌损伤的发病与防治工作。
基金资助:
湖北省中央引导地方科技发展专项（2019ZYYD065）; 湖北省重点实验室开放项目（2021KFY023）

Study on mechanism of isoliensinine reducing acute lung injury induced by lipopolysaccharide through inhibiting TLR4

LI Wen^a, SONGYANG Yiyan^b, SONG Jinchun^a

a Department of Emergency,b Department of Pharmacy,Renmin Hospital of Wuhan University,Wuhan Hubei 430060,China

Received:2023-04-11 Revised:2023-05-09 Online:2023-11-15 Published:2026-03-26
Contact: SONGYANG Yiyan,Deputy chief pharmacist,E-mail：919572095@qq.com

摘要/Abstract

摘要： 目的探讨异莲心碱对脂多糖（lipopolysaccharide,LPS）诱导的急性肺损伤的作用和作用机制。方法 C57BL6J小鼠随机分为4组：对照组、模型组（LPS组）、异莲心碱组和LPS+异莲心碱组,每组12只。小鼠气管内注射200 μg LPS诱导急性肺损伤模型;采用苏木精伊红染色（H&E）、肺脏湿重/干重比值和炎症指数评价急性肺炎程度;采用ELISA试剂盒检测促炎症因子肿瘤坏死因子（tumor necrosis factor-α,TNF-α）、白细胞介素-1（interleukin-1,IL-1）和白细胞介素-6（interleukin-6,IL-6）的水平;采用试剂盒检测活性氧（reactive oxygen species,ROS）、丙二醛（malondialdehyde,MDA）、锰超氧化物歧化酶（superoxide dismutase,SOD2）和谷胱甘肽还原酶（glutathione reductase,Gpx）的水平/活性;采用Western Blot检测相关信号分子的改变。结果 H&E染色显示,LPS组肺组织炎症浸润明显增多,肺脏湿重/干重比值（4.09±0.55vs5.73±0.55）和炎症指数高于对照组（0.36±0.11vs4.25±0.53）;ELISA检测结果显示,LPS组肺脏组织中TNF-α[（45.93±8.08）vs（198.71±37.85）pg/mL]、IL-1[（111.31±6.54）vs（151.27±18.39）pg/mL]和IL-6[（50.15±11.72）vs（468.07±41.23）pg/mL]的水平明显高于对照组;LPS组ROS（1.00±0.23vs4.73±0.39）和MDA含量（1.00±0.09vs4.53±0.07）高于对照组,SOD2[（2.07±0.11）vs（0.74±0.22）U/mg]和Gpx[（188.03±16.64）vs（69.59±13.51）μmol/g]的活性明显低于对照组。LPS+异莲心碱组肺脏湿重/干重比值（4.88±0.39）、炎症指数（3.05±0.49）、炎症因子、ROS（2.31±0.32）和MDA含量（2.97±0.47）均低于LPS组;但是SOD2[（1.65±0.13）U/mg]和Gpx的活性[（143.61±24.21）μmol/g]明显高于LPS组。Western Blot结果显示,LPS组肺脏组织中toll样受体（toll like receptor 4,TLR4）显著上调,磷酸化的核因子-κB（nuclear factor-κB,NF-κB）明显增加;而LPS+异莲心碱组肺脏组织中TLR4和P-NF-κB的表达低于LPS组。结论异莲心碱可以通过抑制TLR4减轻脂多糖诱导的急性肺损伤。异莲心碱可能成为治疗急性肺损伤的新药物。

关键词: 异莲心碱, 脂多糖, 急性肺损伤, Toll 样受体4

Abstract: Objective To explore the effect and mechanism of isoliensinine on lipopolysaccharide（LPS）-induced acute lung injury. Methods C57BL6J mice were randomly divided into four groups：control group,model group（LPS group）,isoliensinine group,LPS+isoliensinine group,12 animals in each group. The mice were treated with intratracheal injection of 200 μg LPS to induce acute lung injury model. The H&E staining,lung wet weight/dry weight ratio and inflammation index were used to evaluate the degree of acute pneumonia. ELISA kit was used to detect pro-inflammatory factors,tumor necrosis factor TNF-α（TNF-α）,interleukin-1（IL-1） and interleukin-6（IL-6） levels. The kits were used to detect reactive oxygen species（ROS）,malondialdehyde （MDA）,superoxide dismutase（SOD） and glutathione reductase（Gpx） levels/activity. The Western blot was used to detect changes in related signal molecules. Results H&E staining showed that the inflammatory infiltration of the lung tissues in the LPS group increased,and the wet weight/dry weight ratio（4.09±0.55vs5.73±0.55） and inflammation index（0.36±0.11vs4.25±0.53） of the lungs were higher than those of the control group. The ELISA results showed that the levels of TNF-α[（45.93±8.08）vs （198.71±37.85）pg/mL],IL-1[（111.31±6.54）vs（151.27±18.39）pg/mL] and IL-6[（50.15±11.72）vs（468.07±41.23）pg/mL] in the lung tissues of the LPS group were significantly higher than those of the control group,the content of ROS（1.00±0.23vs4.73±0.39）and MDA（1.00±0.09vs4.53±0.07） in the LPS group was higher than those of the control group,and the activities of SOD2 [（2.07±0.11）vs（0.74±0.22）U/mg] and Gpx[（188.03±16.64）vs（69.59±13.51）μmol/g] were significantly lower than those of the control group. The lung wet weight/dry weight ratio（4.88±0.39）,inflammation index（3.05±0.49）,inflammatory factors,and contents of ROS（2.31±0.32） and MDA（2.97±0.47） in LPS+isoliensinine group were lower than those in LPS group,while the activities of SOD2[（1.65±0.13）U/mg] and Gpx[（143.61±24.21）μmol/g] were significantly higher than those of LPS group. The Western blot results showed that toll like receptor 4（TLR4） and phosphorylated nuclear factor-κB（NF-κB） of lung tissues in LPS group were significantly increased,while the expression of TLR4 and phosphorylated NF-κB of LPS+isoliensinine group was lower than those of LPS group. Conclusion The isoliensinine can reduce acute lung injury induced by lipopolysaccharide by inhibiting TLR4. The isoliensinine may be a new treatment for lung injury.

Key words: Isoliensinine, Llipopolysaccharide, Acute lung injury, Toll-like receptor 4

中图分类号:

R-332

李文, 宋杨一嫣, 宋金春. 异莲心碱通过抑制TLR4减轻脂多糖诱导的急性肺损伤作用机制的研究. [J]职业与健康, 2023, 39(22): 3067-3071.

LI Wen, SONGYANG Yiyan, SONG Jinchun. Study on mechanism of isoliensinine reducing acute lung injury induced by lipopolysaccharide through inhibiting TLR4. [J]OCCUPATION AND HEALTH, 2023, 39(22): 3067-3071.

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