基于TLR4/NF-κB信号通路研究木犀草素对幼鼠急性肺损伤的作用机制  

Study on the Mechanism of Luteolin on Acute Lung Injury in Young Rats Based on TLR4/NF-κB Signaling Pathway

作　　者：邹国涛[1] 王英娟[2] 曾毅文 张伟[1] ZOU Guotao;WANG Yingjuan;ZENG Yiwen;ZHANG Wei(Department of Pediatrics,Yongchuan Hospital,Chongqing Medical University,Chongqing 402160,China;Emergency Department,Yongchuan Hospital,Chongqing Medical University,Chongqing 402160,China)

机构地区：[1]重庆医科大学附属永川医院儿科,重庆402160 [2]重庆医科大学附属永川医院急诊科,重庆402160

出　　处：《中药新药与临床药理》2021年第5期661-666,共6页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：2019年重庆市教委项目(KJQN201900420)。

摘　　要：目的研究木犀草素对幼鼠急性肺损伤(ALI)的保护作用,并探究其作用机制。方法将50只SPF级SD幼鼠随机分为5组:正常组,模型组,地塞米松组,木犀草素高、低剂量组,每组10只。除正常组外,其余各组幼鼠雾化吸入脂多糖(LPS)建立急性肺损伤模型,正常组大鼠雾化吸入等量生理盐水。模型复制前3 d起及模型复制后1 h,木犀草素高、低剂量组大鼠分别灌胃40、20 mg·kg^(-1)的木犀草素,地塞米松组于模型复制前2 h注射5 mg·kg^(-1)地塞米松,正常组和模型组大鼠灌胃等量0.5%羧甲基纤维素钠(CMC-Na)。模型复制后12 h,采用ELISA法检测血清及肺泡灌洗液(BALF)中白细胞介素1β(IL-1β)、白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)和肺组织超氧化物歧化酶(SOD)活力、丙二醛(MDA)含量;检测大鼠肺组织湿干质量比(W/D);HE染色检测肺组织病理变化;Western Blot法和qRT-PCR法检测肺组织Toll样受体4(TLR4)、核因子κB(NF-κB)p65蛋白和mRNA的表达水平。结果与正常组比较,模型组血清及BALF中IL-1β、IL-6、TNF-α的表达水平明显升高,W/D值升高,肺组织SOD活力降低,MDA含量升高,肺损伤明显,TLR4、NF-κB p65蛋白和mRNA的表达水平增加(P<0.05)。与模型组比较,木犀草素降低了血清及BALF中IL-1β、IL-6、TNF-α水平和W/D值,提高了肺组织SOD活力,降低了MDA含量,肺损伤得到改善,同时也降低了肺组织中TLR4、NF-κB p65蛋白和mRNA的表达水平(P<0.05)。结论木犀草素对幼鼠急性肺损伤具有保护作用,其作用机制与抑制TLR4/NF-κB信号通路的活化有关。Objective To study the protective effect of luteolin on acute lung injury(ALI)in young rats and exploreits mechanism.Methods50 SPF SD young rats were randomly divided into 5 groups:normal group,model group,dexamethasone group,luteolin high-dose group and low-dose group,10 rats in each group.Except for the normalgroup,young rats in the other groups were used to establish ALI model by atomizing LPS,and the rats in thenormal group inhaled the same amount of normal saline.Three days before modeling and 1 hour after modeling,ratsin the luteolin high-dose group and low-dose group were given intragastrically 40 mg·kg^(-1) and 20 mg·kg^(-1) luteolin,respectively.The dexamethasone group was given a one-time injection of 5 mg·kg^(-1) dexamethasone 2 h beforemodeling.Rats in normal group and model group were given 0.5%carboxymethylcellulose sodium(CMC-Na)byintragastric administration.12 h after modeling,ELISA method was used to detect the levels of interleukin-1β(IL-1β),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α)in serum and alveolar lavage fluid(BALF),superoxide dismutase(SOD)activity and malondialdehyde(MDA)in lung tissues.The wet-to-dry weight ratio(W/D)of rat lung tissues was detected.HE staining was used to detect pathological changes of lung tissues.Western blotand q RT-PCR methods were used to detect the expression levels of toll-like receptor 4(TLR4),nuclear factorkappa-B(NF-κB)p65 protein and m RNA in lung tissues.ResultsCompared with normal group,the expressionlevels of IL-1β,IL-6,and TNF-αin the serum and BALF of model group increased significantly,the W/D valueincreased,the SOD activity of the lung tissues decreased,the MDA content increased,and the lung injury wasobvious.The expression levels of TLR4,NF-κB p65 protein and m RNA increased(P<0.05).Compared with modelgroup,luteolin reduced IL-1β,IL-6,TNF-αlevels in serum and BALF,lowered the W/D value,increased lungtissues SOD activity,reduced MDA content,and improved lung injury.It also reduced the expression levels ofTLR4,NF-κB p65 protein a

关 键 词：急性肺损伤 木犀草素 TLR4/NF-κB信号通路 幼鼠 

分 类 号：R285.5[医药卫生—中药学]