The Roots Of Ilex Asprella Extract Lessens Acute Lung Injury In Mice Induced By Influenza Virus

ObjectiveThe plant Ilex asprella(Hook.&Arn.)Champ,ex Benth.,which is one of the native herbal medicines in South China,is mainly distributed in the Guangdong,Guangxi and Fujian provinces.Its roots and leaves have been a common part of herbal tea in treating influenza,bacterial infections and lung abscess for hundreds of years.Previous experiments revealed that it mainly comprises triterpenoid saponin,phenolic acid and alkaloid phenolic compounds.It has been demonstrated that IA extract has significant anti-inflammatory and anti-influenza activity.This paper aims to elucidate its treatment effect and the mechanism of antiviral and anti-inflammatory of acute lung injury in mice caused by the H1N1 virus,and its compounds effects of antiviral and anti-inflammatory in vitro.Methods1.The Ribavirin was used as positive control drug.After being inoculated with several viral doses of influenza A/FM/1/47 H1N1 virus,KM mice were randomly divided into normal control group,model control group,the ribavirin group,high dose group,low dose group of Ilex asprella.Mice survival rate and clinical condition were observed for 15 days after inoculation.Lung index,lung wet/dry weight ratio and pathological analysis were assessed to evaluate the protective effect of lung injury in mice.2.The Ribavirin was used as positive control drug.After being inoculated with several viral doses of influenza A/FM/1/47 H1N1 virus,KM mice were randomly divided into normal control group,model control group,the ribavirin group,high dose group,low dose group of Ilex asprella.The arterial blood gas analysis,pulmonary viral load,alveolar lavage fluid leukocyte count and inflammatory cytokine levels in serum were analyzed to clarify the mechanism of lung injury in mice.3.The Oseltamivir was used as positive control drug.PR8 H1N1 virus was used to infect A549 cell and studied the inhibitory effect of Monomer compounds in NA and IP-10.Results1.The study of protective effects of viral lung injury:compared with model control group,the extract of Ilex asprellta can improve the clinical symptoms of influenza pneumonia in mice,and further reduce the mortality of viral pneumonia in mice(P<0.01).The high dose group can effectively reduce the lung index and wet/dry weight(P<0.01).The high dose group can also reduce the histological changes,an enormous amount of inflammatory cells was less exuded and observed at alveolar and bronchial walls and no severe hemorrhage at alveolar near the bronchioles was observed.2.The mechanism of protective effects of viral lung injury:compared with model control group,the extract of Ilex asprella can improve remarkably increased Pa02 and Sa02 in arterial blood(P<0.05);and decreased lung viral load(P<0.05).Moreover,IL-6,TNF-? and MCP-1 were remarkably decreased in the serum(P<0.01;P<0.05),IL-10 and IFNr-? were remarkably increased in the serum(P<0.01;P<0.05).After oral administration of Ilex asprella extract,the total number of leukocytes in BALF of mice shows a marked decrease(P<0.01).Results showed that there was a sharp decrease in the number of lymphocytes and PMNs in mice(P<0.01;P<0.05),but the number of macrophages was significantly increased(P<0.05).3.The inhibitory effect of influenza virus infection in the A549 cells supernatant of the NA and IP-10:monomer compounds 693 and 925 CC50 were bigger than 100?M.NA inhibition in 693 EC50 was 1.7?M and therapeutic index SI>58.8;NA inhibition EC50 in 925 was 1.7?M and therapeutic index SI>58.8;IP-10 inhibition in 693 EC50 was 2.5?M and therapeutic index SI>40;IP-10 inhibition EC50 in 925 was 6.6?M and therapeutic index SI>15.5.Conclusion1.This study successfully replicate the model of viral lung injury,extract from Ilex asprella has good treatment effect for virus pneumonia,and improve the clinical symptoms of influenza pneumonia in mice,reduce the mortality,can effectively alleviate the virus pulmonary edema and inhibit the further development of lung inflammation.2.The extract from Ilex asprella could increase the oxygen content in the arterial blood,further improve hypoxemia caused by influenza viruses,reduce the mortality of mice;inhibit the replication of influenza virus in the lungs,alleviate the inflammation of the lungs caused by the influenza virus;By inhibiting the shortness of the excessive secretion of inflammatory cytokines,increase the secretion of IL-10 and IFN-y so as to improve the body's antiviral ability,inhibit virus infection mice lung injury;reduce pulmonary lymphocytes and neutrophils,reduce the effect of lung inflammation,increase pulmonary macrophages at the same time the function of lung tissue damage.3.Both compounds from Ilex asprella have inhibitory effect of influenza virus infection in the A549 cells supernatant of the NA and IP-10,and has wide range of security.The result showed that compounds from Ilex asprella could inhibit infected cells from secreting NA and IP-10 in vitro experiments,and have an antiviral and anti-inflammatory effects.