虎杖苷对角叉菜胶致肺水肿大鼠的干预作用研究  

Intervention Effect of Polydatin on Carrageenan-Induced Pulmonary Edema in Rats

作　　者：张钦钦 曾梦楠[1,2,3] 张贝贝 王茹 冯卫生 郑晓珂[1,2,3] ZHANG Qinqin;ZENG Mengnan;ZHANG Beibei;WANG Ru;FENG Weisheng;ZHENG Xiaoke(Henan University of Chinese Medicine,Zhengzhou 450046 Henan,China;The Engineering and Technology Center for Chinese Medicine Development of Henan Province,Zhengzhou 450046 Henan,China;Henan University of Chinese Medicine,Co-construction of Collaborative Innovation Center for Chinese Medicine and Respiratory Diseases by Henan&Education Ministry of P.R.China,Zhengzhou 450046 Henan,China)

机构地区：[1]河南中医药大学,河南郑州450046 [2]河南省中药开发工程技术研究中心,河南郑州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,河南郑州450046

出　　处：《中药新药与临床药理》2023年第7期878-886,共9页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家重点研发计划-中医药现代化研究项目(2019YFC1708802);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(ZYQR201810080)。

摘　　要：目的探讨虎杖苷对角叉菜胶致肺水肿大鼠的干预作用及相关机制。方法将50只SD大鼠随机分为对照组、模型组、地塞米松组(0.075 mg·kg^(-1))及虎杖苷低、高剂量组(20、40 mg·kg^(-1)),每组10只;灌胃给药,每日1次,连续7 d。给药结束后,通过胸腔注射1%角叉菜胶(20 mg·kg^(-1))溶液致炎48 h,建立肺水肿大鼠模型。通过计算肺组织湿/干质量比、肺组织含水量评价肺组织水肿情况;采用HE染色法观察肺组织病理变化;流式细胞术检测肺组织原代细胞凋亡、活性氧(ROS)水平,以及肺组织中巨噬细胞、中性粒细胞水平;检测肺泡灌洗液中免疫球蛋白E(IgE)、干扰素γ(IFN-γ)、白细胞介素4(IL-4)水平,以及肺组织中谷胱甘肽过氧化物酶(GSH-Px)、总超氧化物歧化酶(T-SOD)、丙二醛(MDA)水平;RT-qPCR法检测肺组织中水通道蛋白1(AQP1)、AQP5、IL-10、肿瘤坏死因子α(TNF-α)、IL-13 mRNA表达;免疫组化法测定肺组织中AQP1蛋白表达,并对虎杖苷与AQP1进行分子对接验证;Western Blot法检测肺组织中AQP1、AQP5、IL-1β、IL-10蛋白表达。结果与对照组相比,模型组大鼠的肺组织湿/干质量比及含水量明显升高(P<0.05,P<0.01);肺泡结构破坏及炎症浸润明显;肺组织原代细胞中的活细胞比例显著降低(P<0.01),死细胞、凋亡细胞比例显著升高(P<0.01);肺组织原代细胞ROS水平及肺组织MDA水平显著升高(P<0.01),T-SOD、GSH-Px水平显著降低(P<0.01);肺组织中巨噬细胞、中性粒细胞水平显著升高(P<0.01);肺泡灌洗液中IgE、IL-4水平显著升高(P<0.01),IFN-γ水平显著降低(P<0.01);肺组织AQP1、AQP5、IL-10 mRNA表达水平显著降低(P<0.01),TNF-α、IL-13 mRNA表达水平显著升高(P<0.01);肺组织中AQP1、AQP5、IL-10蛋白表达量显著降低(P<0.01),IL-1β蛋白表达量显著升高(P<0.01)。与模型组比较,虎杖苷高、低剂量组大鼠的肺组织湿/干质量比及含水量明显降低(P<0.05,P<0.01);肺组织的病理�Objective To investigate the interventional effects and related mechanisms of polydatin on carrageenaninduced pulmonary edema in rats.Methods Fifty SD rats were randomly divided into control group,model group,Dexamethasone group(0.075 mg·kg^(-1))and polydatin low-and high-dose groups(20 and 40 mg·kg^(-1)),10 rats in each group;the drug was administered by gavage once a day for 7 consecutive days.After the administration,a rat model of pulmonary edema was established by thoracic injection of 1%carrageenan(20 mg·kg^(-1))solution to cause inflammation for 48 hours.The lung tissue edema was evaluated by calculating the wet/dry mass ratio and the water content of lung tissue;the pathological changes of lung tissue were observed by HE staining;the levels of apoptosis and reactive oxygen species(ROS)in primary cells,and the levels of macrophages and neutrophils in lung tissue were detected by flow cytometry;the levels of immunoglobulin E(IgE),interferonγ(IFN-γ)and leukocyte 4(IL-4)and the levels of glutathione peroxidase(GSH-Px),total superoxide dismutase(T-SOD)and malondialdehyde(MDA)in lung tissue were measured by RT-qPCR;the mRNA expressions of aquaporin 1(AQP1),AQP5,IL-10,tumor necrosis factorα(TNF-α)and IL-13 in lung tissue were measured by immunohistochemistry.Protein expression of AQP1 in lung tissue was determined by immunohistochemistry and validated by molecular docking of polydatin with AQP1;protein expressions of AQP1,AQP5,IL-1β,IL-10 in lung tissue were detected by Western Blot.Results Compared with the control group,the lung tissue wet/dry mass ratio and water content of the model rats were significantly increased(P<0.05,P<0.01);alveolar structural destruction and inflammatory infiltration were significant;the proportion of live cells in primary lung tissue was significantly decreased(P<0.01),and the proportion of dead and apoptotic cells was significantly increased(P<0.01);the levels of ROS in primary lung tissue and MDA in lung tissue were significantly increased(P<0.01)and T-SOD and GSH-Px level

关 键 词：虎杖苷 肺水肿 炎症反应 氧化应激 细胞凋亡 水通道蛋白1 大鼠 

分 类 号：R285.5[医药卫生—中药学]