摘要
目的:探究壮药龙盘止咳方的抗流感病毒性肺炎作用及其相关机制。方法:BALB/c小鼠随机分为对照组、模型组、高剂量(10.4 g/kg)龙盘止咳方组、低剂量(5.2 g/kg)龙盘止咳方组和龙盘止咳方(10.4 g/kg)+Toll样受体3(TLR3)激动剂聚肌胞苷酸[Poly(I:C),20 mg/kg]组,每组20只。采用甲型流感病毒(IAV)H1N1滴鼻法建立病毒性肺炎模型,观察小鼠一般状态,并每组取10只小鼠,记录15 d的存活率和存活时间;计算肺、脾和胸腺指数;HE染色观察肺组织病理变化;实时荧光定量PCR检测肺病毒载量;ELISA法检测肺匀浆中白细胞介素6(IL-6)、IL-4、干扰素α(IFN-α)、IFN-β和IFN-γ水平;Western blot检测肺组织TLR3/视黄酸诱导基因I(RIG-I)/核因子κB(NF-κB)通路相关蛋白的表达。结果:对照组小鼠精神状态良好,在实验期间未出现死亡;与对照组相比,模型组小鼠在感染后出现典型的流感症状,小鼠存活率、存活时间、脾指数和胸腺指数显著降低(P<0.05),肺指数,肺组织病理学评分,病毒载量,肺匀浆中IL-6、IL-4、IFN-γ、IFN-α和IFN-β水平,肺匀浆中IFN-γ/IL-4比值,以及肺组织TLR3、胞核NF-κB p65、RIG-I和干扰素β启动子刺激蛋白1(IPS-1)表达均显著增加(P<0.05);与模型组相比,高、低剂量龙盘止咳方组小鼠症状明显减轻,小鼠存活率、存活时间、脾指数、胸腺指数、肺匀浆中IFN-α和IFN-β水平及肺组织RIG-I和IPS-1表达均显著增加(P<0.05),肺指数,肺组织病理学评分,病毒载量,肺匀浆中IL-6、IL-4和IFN-γ水平,肺匀浆中IFN-γ/IL-4比值,以及TLR3和胞核NF-κB p65表达均显著降低(P<0.05);而Poly(I:C)可明显减弱龙盘止咳方对IAV H1N1感染小鼠的肺脏保护作用。结论:壮药龙盘止咳方有减轻IAV H1N1感染小鼠肺损伤的作用,其机制可能与调节TLR3/RIG-I/NF-κB信号通路,抑制过度的先天炎症反应有关。
AIM:To explore the effect of Zhuang medicine Longpan-Zhike formula(LPZKF)on anti-influenza virus pneumonia and its related mechanism.METHODS:BALB/c mice were randomly divided into control group,model group,high-dose(10.4 g/kg)LPZKF group,low-dose(5.2 g/kg)LPZKF group,and LPZKF(10.4 g/kg)+Tolllike receptor 3(TLR3)agonist polyinosinic:polycytidylic acid[Poly(I:C),20 mg/kg]group,with 20 rats in each group.Influenza A virus(IAV)H1N1 intranasal drip method was used to establish a viral pneumonia model.The general state of the mice was observed,and 10 mice were taken from each group to record the 15-day survival rate and survival time.The lung,spleen and thymus indexes were calculated,and HE staining was used to observe the pathological changes of lung tissue.Real-time fluorescence quantitative PCR was used to detect lung viral load,and ELISA was used to measure the levels of interleukin(IL)-6,IL-4,interferon(IFN)-α,IFN-β and IFN-γ in lung homogenates.Western blot was used to detect the expression of TLR3/retinoic acid-inducible gene-I(RIG-I)/nuclear factor-κB(NF-κB)pathway related proteins in lung tissues.RESULTS:All control mice were shown in good mental state and did not die during the experiment.Compared with control group,the mice in model group showed typical flu symptoms after infection,the survival rate,survival time,spleen index and thymus index of the mice were reduced significantly(P<0.05),while the lung index,lung histopathological score,viral load,IL-6,IL-4,IFN-γ,IFN-α and IFN-β levels in lung homogenates,IFN-γ/IL-4 ratio in lung homogenates,and TLR3,nuclear NF-κB p65,RIG-I and interferon-β promoter stimulator 1(IPS-1)expression in lung tissues were increased significantly(P<0.05).Compared with model group,the symptoms of the mice in high-and lowdose LPZKF groups were attenuated significantly,the survival rate,survival time,spleen index,thymus index,IFN-α and IFN-β levels in lung homogenates,and RIG-I and IPS-1 expression in lung tissues were increased significantly(P<0.05),while the lung inde
作者
邹敏
谢彩英
甘娜
农志飞
李崇进
韦杏
ZOU Min;XIE Cai-ying;GAN Na;NONG Zhi-fei;LI Chong-jin;WEI Xing(Department of Pediatrics,Guangxi International Zhuang Medical Hospital,Nanning 530001,China;Zhuang College of Medicine,Guangxi University of Traditional Chinese Medicine,Nanning 530200,China;Department of Pediatrics,the First Affiliated Hospital of Guangxi Medical University,Nanning 530021,China;Department of Pediatrics,Maoming Hospital of Traditional Chinese Medicine,Maoming 525099,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2022年第3期526-534,共9页
Chinese Journal of Pathophysiology
基金
广西高校中青年教师基础能力提升项目(No.2018KY0302)
2021年广西国际壮医医院院级课题(No.GZ2021012)。
