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  

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

作　　者：李文[1] 宋杨一嫣 宋金春[1] LI Wen;SONGYANG Yiyan;SONG Jinchun(Department of Emergency,Renmin Hospital of Wuhan University,Wuhan Hubei,430060,China;Department of Pharmacy,Renmin Hospital of Wuhan University,Wuhan Hubei,430060,China)

机构地区：[1]武汉大学人民医院急诊科,湖北武汉430060 [2]武汉大学人民医院药学部,湖北武汉430060

出　　处：《职业与健康》2023年第22期3067-3071,共5页Occupation and Health

基　　金：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)。

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200μg 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。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 200μg 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.Th

关 键 词：异莲心碱 脂多糖 急性肺损伤 TOLL样受体4 

分 类 号：R-332[医药卫生]