基于NLRP3炎性体及细胞焦亡研究犀角地黄汤对流感病毒性肺炎小鼠炎症反应的抑制作用  

The inhibitory effect of Xijiao Dihuang Decoction on inflammatory response in mice with influenza virus pneumonia based on NLRP3 inflammasome and pyroptosis

作　　者：马璐瑶 杨明锐 邓迪 周娜 刘梦华 吴珺 郝钰 MA Luyao;YANG Mingrui;DENG Di;ZHOU Na;LIU Menghua;WU Jun;HAO Yu(Beijing University of Chinese Medicine,Beijing 102488,China)

机构地区：[1]北京中医药大学生命科学学院,102488

出　　处：《环球中医药》2023年第4期627-634,共8页Global Traditional Chinese Medicine

基　　金：国家自然科学基金(81573723)。

摘　　要：目的探究犀角地黄汤对流感病毒性肺炎小鼠炎症反应的作用及作用机制。方法BALB/c小鼠,随机分为正常组、模型组、犀角地黄汤组、达菲组,除正常组外其余小鼠以流感病毒滴鼻感染,1小时后给药。各组分别以对应的水煎剂或溶液灌胃,每日1次,连续6天。BALB/c小鼠分别在感染后第2、4、6天取材,观察小鼠肺大体病变情况;计算肺指数;光镜下观察肺组织病理切片;RT-PCR法检测肺组织中NOD样受体热蛋白结构域相关蛋白3(NOD-like receptor thermal protein domain associated protein 3,NLRP3)、凋亡相关斑点样蛋白(apoptosis-associated speck-like protein,ASC)、半胱天冬氨酸酶1前体(Pro-Caspase-1)和gasdermin D(GSDMD)mRNA水平;ELISA法检测小鼠血清中肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)、白细胞介素1β(interleukin-1β,IL-1β)和白细胞介素6(interleukin-6,IL-6)水平。Western blot检测肺组织中NLRP3、Caspase-1 p20、GSDMD和gasdermin D N端片段(GSDMD-N)蛋白表达量。结果犀角地黄汤组和达菲组均可在第2、4、6天改善小鼠肺脏充血、红肿和坏死现象,且不同程度的减轻感染小鼠肺组织炎性病理改变。第6天,犀角地黄汤组的肺指数显著降低。模型组小鼠肺组织NLRP3、ASC、Pro-Caspase-1、GSDMD mRNA表达增多,NLRP3、Caspase-1 p20、GSDMD和GSDMD-N蛋白表达亦增多,肺组织IL-1β、IL-6、TNF-α水平升高;犀角地黄汤可显著减少上述分子mRNA、蛋白表达及炎症因子水平。结论犀角地黄汤通过调控NLRP3-Caspase1介导的细胞焦亡抑制流感病毒性肺炎小鼠的炎症反应,是其截断疾病的传变的机制之一。Objective To explore the effect and mechanism of Xijiao Dihuang decoction on inflammatory response in mice with influenza virus pneumonia.Methods BALB/c mice were randomly divided into normal group,model group,Xijiao Dihuang decoction group and Oseltamivir phosphate capsules group.Except the normal group,the other mice were infected with influenza virus by nasal drip,and treated with influenza virus 1 hour later.Each group was treated with the corresponding decoction or solution by gavage once a day for 6 consecutive days.The specimens of BALB/c mice were collected on the 2nd,4th and 6th day after infection,and the gross lung lesions of the mice were observed.Lung index was calculated.The pathological sections of lung tissue were observed under light microscope.RT-PCR was used to measure the mRNA levels of nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3),apoptosis-associated speck-like protein(ASC),Pro-Caspase-1 and gasdermin D(GSDMD)in lung tissue.The serum levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and interleukin-6(IL-6)were detected by ELISA.Western blot was used to detect the protein expression of NLRP3,Caspase-1 p20,GSDMD and gasdermin D N-terminal fragment(GSDMD-N)in lung tissue.Results Both Xijiao Dihuang decoction and Oseltamivir phosphate capsules treatment could improve the lung congestion,redness,swelling and necrosis of mice on the 2nd,4th and 6th day,and alleviate the inflammatory pathological changes of lung tissue in the infected mice to varying degrees.On the 6th day,the lung index of the Xijiao Dihuang decoction group was decreased significantly.The expression of NLRP3,ASC,Pro-Caspase-1 and GSDMD mRNA in lung tissue of the model group was increased,and the protein expression of NLRP3,Caspase-1 p20,GSDMD and GSDMD-N was also increased,and the levels of IL-1β,IL-6 and TNF-α in lung tissue were increased.Xijiao Dihuang decoction can significantly reduce the mRNA and protein expression of the above molecules and the levels of inflammatory factors.Conc

关 键 词：犀角地黄汤 截断疗法 流感病毒 NOD样受体热蛋白结构域相关蛋白3-半胱氨酸蛋白酶1 细胞焦亡 炎症反应 

分 类 号：R285.5[医药卫生—中药学]