沙奎那韦对脂多糖诱导急性肺损伤大鼠PI3K-AKt-NF-κB信号通路的影响  

Effect of saquinavir on PI3K-AKt-NF-κB signaling pathway in rats with acute lung injury induced by lipopolysaccharide

作　　者：何达 张莉 席俊峰[1] He Da;Zhang Li;Xi Junfeng(Department of Respiratory Medicine,The First Hospital of Yulin,Yulin 719000,China)

机构地区：[1]榆林市第一医院呼吸内科,榆林719000

出　　处：《广西医科大学学报》2022年第3期418-423,共6页Journal of Guangxi Medical University

基　　金：陕西省自然科学基础研究计划项目(No.2018JM7068)。

摘　　要：目的:研究沙奎那韦(SQV)对脂多糖(LPS)诱导急性肺损伤(ALI)大鼠的保护作用及其对PI3K-AKt-NF-κB信号通路的影响。方法:将36只雄性SD大鼠随机分为对照组(Control组)、LPS模型组(LPS组)、SQV处理的LPS模型组(LPS+SQV组),每组12只。LPS+SQV组大鼠在LPS造模前予以200 mg/kg·d-1的SQV连续5 d灌胃处理,Control组与LPS组大鼠用等体积的生理盐水灌胃处理,第5天LPS组和LPS+SQV组大鼠采用腹腔注射5 mg/kg的LPS制备ALI模型,造模12 h后腹腔注射120mg/kg戊巴比妥处死大鼠;苏木精—伊红(HE)染色观察各组肺组织病理变化,计算各组大鼠肺组织湿/干(W/D)比值,TUNEL染色对肺组织细胞凋亡水平进行测定,采用ELISA法检测每组大鼠支气管肺泡灌洗液中炎症因子:肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、白细胞介素-10(IL-10)的表达量,Western blotting检测肺组织中PI3K-AKt-NF-κB信号通路相关蛋白IκBa的表达水平和PI3K、AKt、NF-κB p65蛋白的磷酸化水平。结果:与Control组相比,LPS组的大鼠肺组织病理学损伤加重,肺组织W/D比值升高(P<0.05),TNF-α、IL-1β、IL-6、IL-10含量和PI3K的磷酸化水平(p-PI3K/PI3K)、AKt的磷酸化水平(p-AKt/AKt)、NF-κB的磷酸化水平(p-NF-κB p65/NF-κB p65)均升高(P<0.05),而IκBa表达水平降低(P<0.05),细胞凋亡率升高(P<0.01);与LPS组相比,LPS+SQV组的大鼠肺组织病理学损伤减轻,肺组织W/D比值下降(P<0.05),细胞凋亡率降低(P<0.01),IL-10含量、IκBa表达均升高(P<0.05),而其他检测指标均降低(P<0.05)。结论:SQV通过抑制细胞凋亡和炎症反应对LPS诱导的大鼠ALI产生一定的保护作用,其机制可能与PI3K-AK-NF-κB信号通路有关。Objective:To research the protective effect of saquinavir(SQV) on acute lung injury(ALI)-induced by lipopolysaccharide(LPS) and its effect on PI3K-AKt-NF-κB signaling pathway.Methods:A total of 36 male SD rats were randomly divided into Control group,LPS group and LPS+SQV group,with 12 rats in each group.The rats in the LPS+SQV group were treated with SQV(200 mg/kg.d-1,5 days,intragastric administration) before the establishment of the LPS model,while the rats in the Control group and the LPS group were treated with the same volume of normal saline for the same days.On the fifth day,the rats in the LPS group and LPS+SQV group were intra-peritonelly injected with 5 mg/kg LPS to establish the ALI model.Twelve hours after modeling,the rats were euthanized by intraperitoneal injection of 120 mg/kg pentobarbital.The pathological changes of lung tissue in each group were observed by HE staining.The wet/dry(W/D) ratio of lung tissue was measured.The apoptosis rate was detected by TUNEL staining.The expressions of inflammatory factors such as tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),interleukin-6(IL-6) and interleukin-10(IL-10)in bronchoalveolar lavage fluid were determined by ELISA.Western blotting was used to detect the expression of PI3K-AKt-NF-κB signaling pathway related proteins.Results:Compared with the Control group,the pathological injury of lung tissue in LPS group was aggravated;the ratio of W/D ration in lung tissue was increased(P<0.05);the content of TNF-α,IL-1β,IL-6,IL-10,the phosphorylation level of PI3K(p-PI3K/PI3K),the phosphorylation level of AKt(p-AKt/AKt),the phosphorylation level of NF-κB(p-NF-κB p65/NF-κB p65) were increased(P<0.05).However,the expression of IκBa decreased(P<0.05) and the rate of apoptosis increased(P<0.01).Compared with LPS group,the pathological injury of lung tissue in LPS+SQV group decreased,the ratio of W/D decreased(P<0.05),the rate of apoptosis decreased(P<0.01),the content of IL-10 and the expression of IκBa increased(P<0.05),while other indexes dec

关 键 词：沙奎那韦 脂多糖诱导 急性肺损伤 细胞凋亡 炎症反应 PI3K-AKt-NF-κB信号通路 

分 类 号：R285.5[医药卫生—中药学]