热毒宁注射液对甲型流感病毒感染小鼠肺组织炎症和相关炎症因子水平的影响
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摘要
目的观察热毒宁注射液对甲型流感病毒感染小鼠肺组织炎症,以及血清和肺组织肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)表达水平的影响。方法采用滴鼻法建立甲型流感病毒感染小鼠模型。实验设正常对照组、模型组、热毒宁注射液组、利巴韦林注射液组。滴鼻法对小鼠进行流感病毒接种,造模24 h后开始给药5 d,分别于给药后第3、第5日进行取材,计算肺指数,HE染色观察小鼠肺组织病理,ELISA法检测小鼠血清和肺组织TNF-α、IL-1β、IL-6的水平。结果流感病毒感染小鼠后,小鼠肺组织出现炎症改变,肺泡间隔明显增宽,肺指数升高,造模后第5日炎症最为严重。热毒宁注射液组和利巴韦林注射液组肺部炎症有不同程度减轻,热毒宁注射液组作用更明显。流感病毒感染小鼠后,血清和肺泡灌洗液TNF-α、IL-1β、IL-6水平明显增高,在第3日达到高峰;热毒宁注射液和利巴韦林注射液组各时点血清和肺组织TNF-α、IL-1β、IL-6水平均较模型组降低(P <0.05);热毒宁注射液组各时点血清和肺组织TNF-α水平较利巴韦林注射液组降低更明显(P <0.05);治疗第3日,热毒宁注射液组肺组织IL-1β、IL-6水平较利巴韦林注射液组降低更明显(P <0.05)。结论热毒宁注射液可减轻流感病毒感染后的肺部炎症,降低炎症因子的释放。
Objective: To observe the effects of Reduning Injection on lung inflammation and the expression levels of TNF-α,IL-1β and IL-6 in serum and lung tissues of mice infected with influenza A virus. Methods:Rat models infected with Influenza A virus were constructed with nasal dripping method. 4 groups were set up :the normal group,the model group,Reduning Injection group and Ribavirin Injection group. The rats were inoculated with Influenza A virus through nasal dripping. After 24 hours,the rats were given saline intraperitoneally and given medicine orally for 5 days. The lung index was calculated on the 3 rd and 5 th day after administration.Lung histopathology was observed by HE staining. The levels of TNF-α,IL-1β and IL-6 in serum and lung tissue were detected by ELISA. Results: After influenza virus infection,the lung tissue of mice showed inflammation changes: alveolar septum significantly widened,lung index increased,and the inflammation was the most serious on the fifth day after model establishment. Reduning Injection group and Ribavirin Injection group had different degrees of relief of pulmonary inflammation,and Reduning Injection group had more obvious effects. After influenza virus infection,the levels of TNF-α,IL-1β and IL-6 in serum and alveolar lavage fluid of mice increased significantly,reaching the peak on the third day. Those of Reduning Injection group and Ribavirin Injection group decreased compared with the model group(P < 0.05). The levels of TNF-a in serum and lung tissues of Reduning Injection group were significantly lower than those of Ribavirin Injection group(P < 0.05). On the third day,the levels of IL-1β and IL-6 in serum and lung tissues of Reduning Injection group were significantly lower than those of Ribavirin Injection group(P < 0.05). Conclusion: Reduning Injection can alleviate pulmonary inflammation and reduce the release of inflammatory factors after influenza virus infection.
Objective: To observe the effects of Reduning Injection on lung inflammation and the expression levels of TNF-α,IL-1β and IL-6 in serum and lung tissues of mice infected with influenza A virus. Methods:Rat models infected with Influenza A virus were constructed with nasal dripping method. 4 groups were set up :the normal group,the model group,Reduning Injection group and Ribavirin Injection group. The rats were inoculated with Influenza A virus through nasal dripping. After 24 hours,the rats were given saline intraperitoneally and given medicine orally for 5 days. The lung index was calculated on the 3 rd and 5 th day after administration.Lung histopathology was observed by HE staining. The levels of TNF-α,IL-1β and IL-6 in serum and lung tissue were detected by ELISA. Results: After influenza virus infection,the lung tissue of mice showed inflammation changes: alveolar septum significantly widened,lung index increased,and the inflammation was the most serious on the fifth day after model establishment. Reduning Injection group and Ribavirin Injection group had different degrees of relief of pulmonary inflammation,and Reduning Injection group had more obvious effects. After influenza virus infection,the levels of TNF-α,IL-1β and IL-6 in serum and alveolar lavage fluid of mice increased significantly,reaching the peak on the third day. Those of Reduning Injection group and Ribavirin Injection group decreased compared with the model group(P < 0.05). The levels of TNF-a in serum and lung tissues of Reduning Injection group were significantly lower than those of Ribavirin Injection group(P < 0.05). On the third day,the levels of IL-1β and IL-6 in serum and lung tissues of Reduning Injection group were significantly lower than those of Ribavirin Injection group(P < 0.05). Conclusion: Reduning Injection can alleviate pulmonary inflammation and reduce the release of inflammatory factors after influenza virus infection.
引文
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[18]王变利,高燕,赵秀香.金银花水提物体外抗病毒实验研究[J].辽宁中医杂志,2015,42(8):1495-1497.
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[2] Influenza(Seasonal):Fact Sheet. World Health Organization[OL]. http://www.who.int/mediacentre/factsheets/fs211/en/Accessed May 23,2017.
[3] Iuliano AD,Jang Y,Jones J,et al. Increase in human infections with avian influenza A(H7N9)virus during the fifth epidemic-China,October 2016-February 2017[J]. MMWR Morb Mortal Wkly Rep,2017,66(9):254-255.
[4] Liu Y,Mu W,Xiao W,et al. Efficacy and safety of Re-DuNing injection in the treatment of seasonal influenza:results froma randomized,double-blinded,multicenter,oseltamivircontrolled trial[J]. Oncotarget,2017,8(33):55176-55186.
[5] Allen IC,Scull MA,Moore CB,et al. The NLRP3 inflammasome mediates in vivo innate immunity to influenza A virus through recognition of viral RNA[J]. Immunity,2009,30(4):556-565.
[6] Houde M,Arora DJ. Stimulation of tumor necrosis factor secretion by purified influenza virus neuraminidase[J]. Cell Immunol,1990(129):104-111.
[7] Gong JH,Sprenger H,Hinder F,et al. Influenza A virus infection of macrophages:enhanced tumor necrosis factor alpha(TNF alpha)gene expression and lipopoly saccharide-triggered TNF aplha release[J]. J Immunol,1991(147):3507-3513.
[8] Wolkow A,Aisbett B, Jefferies S,et al. Effect of heat exposure and simulated physical firefighting work on acute inflammatory and cortisol responses[J]. Ann Work Expo Health,2017(185):282-288.
[9]王晓晨,吉爱国. NF-κB信号通路与炎症反应[J].生理科学进展,2014,45(1):68-71.
[10] Peper RL,Vancampen H. Tumor necrosis factor as a mediator of inflammation in influenza A virus pneumonia[J]. Mierob Pathog,1995(19):175-183.
[11] Kaiser L,Fritz RS,Straus SE,et al. Symptom pathogenesis during acute influenza:interleukin-6 and other cytokine response[J]. J Med Virol,2001(64):262-268.
[12] Wiley JA,Cervenka A,Harkema JR. Production of interferongamma by influenza hemagglutinin-special CD8 effector T cells influences the development of pulmomary immuno pathology[J]. Am J Pathol,2001(158):119-130.
[13] Hussell T,Pennycook A,Openshaw PJ. Inhibition of tumor necrosis factor reduces the sev erity of virus-specific lung immunopathology[J]. Eur J Immunol,2001(31):2566-2573.
[14]卫生部.卫生部办公厅关于印发《甲型H1N1流感诊疗方案(2010年版)》的通知[OL]. http://www.moh.gov.cn/mohyzs/s3586/201005/47250. Shtml,2010-05-07.
[15]何雄,叶雨华.热毒宁注射液对外感风热证患者炎性因子及炎性介质的影响[J].临床医学工程,2018,25(7):851-852.
[16] Li H,Yu Y,Wang Z,et al. Chemical profiling of Re-Du-Ning injection by ultra-performance liquid chromatography coupled with electrospray ionization tandem quadrupole time-of-flight mass spectrometry through the screening of diagnostic Ions in MS mode[J]. PLoS One,2015(10):e0121031.
[17] Chen Cheng,Feifei Du,Ke Yu,et al. Pharmacokinetics and disposition of circulating iridoids and organic acids in rats intravenously receiving ReDuNing injection[J]. DRUG METAB DISPOS,2016,44(11):1853-1858.
[18]王变利,高燕,赵秀香.金银花水提物体外抗病毒实验研究[J].辽宁中医杂志,2015,42(8):1495-1497.
[19]宋亚玲,王红梅,倪付勇,等.金银花中酚酸类成分及其抗炎活性研究[J]. 2015,46(4):490-495.
[20]张耘实.栀子苷対甲型H1NI流感病毒抑制效应的实验研究[D].南京:南京医科大学,2016.