基于NF-κB/AQPs信号通路探究月见草对肺水肿模型大鼠的预防作用  

Study on the Preventive Effect of Evening Primrose on Pulmonary Edema Rats Based on NF-κB/AQPs Signaling Pathway

作　　者：王茹 曾梦楠[1,2,3] 张贝贝 张钦钦 张宇涵 冯卫生 郑晓珂[1,2,3] WANG Ru;ZENG Meng-nan;ZHANG Bei-bei;ZHANG Qin-qin;ZHANG Yu-han;FENG Wei-sheng;ZHENG Xiao-ke(School of Pharmacy,Henan University of Chinese Medicine,Zhengzhou 450046,China;The Engineering and Technology Center for Chinese Medicine Development of Henan Province,Zhengzhou 450046,China;Collaborative Innovation Center for Chinese Medicine and Respiratory Diseases by Henan Province,Henan University of Chinese Medicine,Zhengzhou 450046,China)

机构地区：[1]河南中医药大学药学院,郑州450046 [2]河南省中药开发工程技术研究中心,郑州450046 [3]河南中医药大学,呼吸疾病中医药防治省部共建协同创新中心,郑州450046

出　　处：《中国药学杂志》2023年第8期689-698,共10页Chinese Pharmaceutical Journal

基　　金：国家重点研发计划-中医药现代化研究专项资助(2017YFC1702800,2019YFC1708802);河南省重大科技专项资助(171100310500);河南省高层次人才特殊支持计划“中原千人计划”-中原领军人才项目资助(ZYQR201810080)。

摘　　要：目的 探究月见草对角叉菜胶诱导肺水肿模型大鼠的预防作用,为临床治疗肺水肿提供参考。方法 将50只SD大鼠随机分为正常组(CON)、模型组(FSZ,20 mg·kg^(-1))、地塞米松组(DEX,0.075 mg·kg^(-1)·d^(-1))、低剂量月见草组(YJC-L,1.75 g·kg^(-1)·d^(-1))以及高剂量月见草组(YJC-H,7 g·kg^(-1)·d^(-1)),每组10只,给药7 d后胸腔注射10 mg·mL^(-1)角叉菜胶致炎建立肺水肿模型。致炎48 h后,计算肺含水量与湿干比;苏木素-伊红(hematoxylin-eosin staining, HE)染色观察肺组织病理学变化;酶联免疫吸附(enzyme linked immunosorbent assay, ELISA)法检测肺泡灌洗液中免疫球蛋白E(immunoglobulin E,IgE)、白细胞介素4(interleukin 4,IL-4)和干扰素-γ(interferon-γ,IFN-γ)的水平;原位末端转移酶标记技术(TUNEL)染色检测肺组织细胞凋亡情况;流式细胞术检测肺组织细胞悬液活性氧(reactive oxygen species, ROS)以及巨噬细胞和中性粒细胞水平;酶活检测肺组织中谷胱甘肽过氧化物酶(glutathione peroxidase, GSH-Px)、总超氧化物歧化酶(total-superoxide dismutase, T-SOD)和丙二醛(malondialdehyde, MDA)的水平;实时荧光定量PCR(qRT-PCR)检测肺组织中水通道蛋白1(aquaporin 1,AQP1)、水通道蛋白5(aquaporin 5,AQP5)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素6(interleukin 6,IL-6)、白细胞介素13(interleukin 13,IL^(-1)3)及白细胞介素10(interleukin 10,IL^(-1)0)的mRNA表达水平;免疫组化检测AQP1的表达水平;Western blot检测肺组织中AQP1、AQP5、磷酸化核转录因子-κB(p-NF-κB p65)及核转录因子-κB(NF-κB p65)蛋白水平。结果 月见草显著降低肺水肿模型大鼠肺组织含水量和湿干比,改善肺组织病理变化;降低肺泡灌洗液中IgE、IL-4的水平及升高IFN-γ的水平,降低肺组织中TNF-α、IL-6、IL^(-1)3和升高IL^(-1)0的mRNA水平;降低肺组织细胞凋亡数目;抑制氧化应激反应;降低肺组织巨噬细胞和中性粒细胞的水平,�OBJECTIVE To investigate the preventive effect of evening primrose on pulmonary edema model induced by carrageenan in rats and provide references for clinical treatment of pulmonary edema.METHODS Fifty SD rats were randomly divided into normal group(CON),model group(FSZ,20 mg·kg^(-1)),dexamethasone group(DEX,0.075 mg·kg^(-1)·d^(-1)),low-dose evening primrose group(YJC-L,1.75 g·kg^(-1)·d^(-1))and high-dose evening primrose group(YJC-H,7g·kg^(-1)·d^(-1)),with 10 rats in each group.Seven days after drug administration,1%carrageenan(2 mL·kg^(-1))was injected into thoracic cavity to induce inflammation and establish pulmonary edema model.After 48 h of inflammation,lung water content and wet-dry ratio were calculated.Hematoxylin-eosin staining(HE)staining was used to observe the pathological changes of lung tissue.The levels of immunoglobulin E(IgE),interleukin 4(IL-4)and interferon-γ(IFN-γ)in bronchoalveolar lavage fluid(BALF)were determined by enzyme linked immunosorbent assay(ELISA).TUNEL staining was used to detect the apoptosis of lung tissue.The levels of reactive oxygen species(ROS),macrophages,and neutrophils in lung tissue cell suspensions were detected by flow cytometry.The levels of glutathione peroxidase(GSH-Px),superoxide dismutase(T-SOD)and malondialdehyde(MDA)in lung tissue were detected by enzyme activity.qRT-PCR was used to detect the mRNA expression levels of aquaporin 1(AQP1),aquaporin 5(AQP5),tumor necrosis factor-α(TNF-α),interleukin 6(IL-6),interleukin 13(IL^(-1)3)and interleukin 10(IL^(-1)0)in lung tissue.The expression level of AQP1 was detected by immunohistochemistry.The protein levels of AQP1,AQP5,phosphorylated nuclear factor kappa B(p-NF-κB p65),and nuclear factor kappa B(NF-κB p65)in lung tissues were detected by Western blot.RESULTS Evening primrose significantly reduced the water content and wet-dry ratio of lung tissue in pulmonary edema model rats and improved the pathological changes of lung tissue.It reduced the levels of IgE and IL-4 and increased the levels of IFN-γi

关 键 词：月见草 肺水肿 炎症 氧化应激 凋亡 水通道蛋白 

分 类 号：R966[医药卫生—药理学]