姜黄素通过影响线粒体跨膜电位和TLR4/NF-κB信号通路减轻LPS诱导急性肺损伤  

Curcumin Alleviates LPS-Induced Acute Lung Injury by Regulating Mitochondrial Transmembrane Potential and TLR4/NF-κB Signaling Pathway

作　　者：赵浩 黄江 陈圣东 陈修瑞 张婉婷 章卓 ZHAO Haoi;HUANG Jiang;CHEN Shengdong;CHEN Xiurui;ZHANG Wanting;ZHANG Zhuo(Department of Pharmacy,Affiliated Traditional Chinese Medicine Hospital of Southwest Medical University,Luzhou 646000;School of Pharmacy,Southwest Medical University,Luzhou 646000)

机构地区：[1]西南医科大学附属中医医院药剂科,泸州646000 [2]西南医科大学药学院,泸州646000

出　　处：《中药药理与临床》2025年第2期33-39,共7页Pharmacology and Clinics of Chinese Materia Medica

基　　金：2023年度四川省中医药管理局中医药科研专项课题面上项目(编号:2023577);泸州市科技局项目(编号:2018LZXNYD-ZK46)。

摘　　要：目的:观察姜黄素是否通过影响线粒体跨膜电位和TLR4/NF-κB信号通路抑制脂多糖(LPS)诱导的急性肺损伤(ALI)。方法:将RAW264.7细胞分为空白对照组、模型对照组、姜黄素5、10、20μmol/L组、地塞米松10μmol/L组,采用10 ng/mL LPS刺激细胞,CCK-8法检测姜黄素对RAW264.7细胞活性的影响;Annexin V-FITC/PI染色检测细胞凋亡率;JC-1染色检测线粒体跨膜电位;Western blot法检测Toll样受体4(TLR4)、核转录因子κB(NF-κB)、BCL-2、BAX蛋白表达;ELISA法检测上清液中肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、IL-6和IL-10含量。将昆明种小鼠72只分为假手术对照组、模型对照组、姜黄素50、100、200 mg/kg组、地塞米松10 mg/kg组,采用50 mg/mL LPS气管滴注造ALI模型,其中36只小鼠用于检测肺湿质量/干质量比;HE染色检测肺组织病理变化;Western blot法检测肺组织TLR4、NF-κB蛋白表达;剩余36只小鼠通过ELISA法检测支气管肺泡灌洗液中TNF-α、IL-6、IL-1β和IL-10含量。结果:与空白对照组比较,模型对照组LPS诱导的RAW264.7细胞存活率显著降低、凋亡率显著增加(P<0.01),BCL-2蛋白表达下调,BAX蛋白表达上调,BAX/BCL-2比值升高(P<0.01),线粒体跨膜电位显著降低(P<0.01),TLR4/NF-κB蛋白表达显著上调(P<0.01),TNF-α、IL-6、IL-1β含量显著增加,IL-10含量显著降低(P<0.01);与模型对照组比较,姜黄素5、10、20μmol/L组RAW264.7细胞存活率提高(P<0.05),细胞凋亡显著降低(P<0.01),BAX/BCL-2比值明显降低(P<0.05),细胞线粒体跨膜电位升高(P<0.05),TLR4/NF-κB蛋白表达下调(P<0.05),TNF-α、IL-6、IL-1β含量降低(P<0.01),IL-10含量增加(P<0.05)。与假手术对照组比较,LPS诱导的小鼠肺病理损伤和肺湿/干质量比显著增加(P<0.01),小鼠肺组织TLR4/NF-κB蛋白表达显著上调(P<0.01),肺泡灌洗液中TNF-α、IL-6、IL-1β含量显著增加(P<0.01),IL-10含量明显降低(P<0.01);与模型对照组比较,姜黄素各组小鼠LPS诱Objective:To investigate whether curcumin(Cur)inhibits lipopolysaccharide(LPS)-induced acute lung injury(ALI)by regulating the mitochondrial transmembrane potential(MMP)and Toll-like receptor 4(TLR4)/nuclear factor(NF)-κB signaling pathway.Methods:RAW264.7 cells were classified into blank control,model,Cur(5,10,and 20μmol/L),and dexamethasone(Dex,10μmol/L)groups.The cells were simulated by 10 ng/mL LPS.The viability,apoptosis,and MMP of RAW264.7 cells were measured by the cell counting kit-8(CCK-8)assay,Annexin V-FITC/PI staining,and JC-1 staining,respectively.The protein levels of TLR4,NF-κB,B cell lymphoma-2(BCL-2),and Bcl-2-associated protein X(BAX)were determined by Western blotting.The levels of tumor necrosis factor(TNF)-α,interleukin(IL)-6,IL-1β,and IL-10 in the supernatant were measured by enzyme-linked immunosorbent assay(ELISA).In the animal experiment,72 Kunming mice were assigned into sham,model,Cur(50,100,and 200 mg/kg),and Dex(10 mg/kg)groups.The mouse model of ALI was constructed by intratracheal instillation of 50 mg/mL LPS.Among them,36 mice were used to measure the lung wet/dry ratio.The patho-logical changes in the lung tissue were observed by hematoxylin-eosin staining,and the protein levels of TLR4 and NF-κB in the lung tissue were determined by Western blotting.The levels of TNF-α,IL-6,IL-1β,and IL-10 in the bronchoalveolar lavage fluid of the remaining 36 mice were measured by ELISA.Results:Compared with the blank control group,the modeling decreased the survival rate and increased the apoptosis rate of RAW264.7 cells(P<0.01),increased the BAX/BCL-2 ratio by down-regulating the expression of BCL-2 and up-regula-ting the expression of BAX(P<0.01),decreased MMP(P<0.01),up-regulated the protein levels of TLR4/NF-κB(P<0.01),elevated the levels of TNF-α,IL-6,and IL-1β(P<0.01),while lowered the level of IL-10(P<0.01)in RAW264.7 cells.Compared with the model group,Cur increased the survival rate(P<0.05)and decreased the apoptosis rate(P<0.01)of RAW264.7 cells,reduced the BAX/BCL-2 rati

关 键 词：姜黄素 急性肺损伤 线粒体跨膜电位 Toll样受体4 核因子-ΚB 凋亡 

分 类 号：R285.5[医药卫生—中药学]