Research On The Traditional Chinese Medicine Depestia With The Effect Of Anti-Influenza Virus

Objective:Diacolation technology using dialysis and ion exchange technology have antiviral activity of components in Commelina communis L alkaloids to the greatest degree of extraction,prepared by Depestia agents.Separately for the in vivo and in vitro anti-influenza virus research,in vitro in MDCK cells was studied and in vivo in Kunming mice was studied to assess the efficacy of Depestia.Method:1.proliferation and titration of virus:influenza A virus using chicken embryo and MDCK cells to determine virus hemagglutination titer experiments,MDCK cells TCID₅₀ virus titration.2.Depestia in vitro antiviral experiment:Depestia by the mother liquor has been diluted to half concentration gradient.Determination of MTT cytotoxic drugs pharmacodynamics experiments at non-toxic concentration range of 96 cells in the plate by adding different concentrations of drug 0.1ml,Liquid Absorption 100TCID₅₀ the virus 0.1ml;the use of the treatment group and prevention group of two to medicine the way,the daily observation of virus-induced cytopathic effect,virus control group about cell + + + CPE when measured by MTT staining 570nm Office of the hole and calculate the OD value of half inhibitory concentration(half effective concentration) is the therapeutic index drugs.3.Depestia in vivo antiviral experiment: Intranasal influenza virus infection in mice to establish animal models of virus infection.Protective effect of the death:the 60 mice(SPF grade,13 - 16g, were equally divided between male and female) were randomly divided into 6 groups of 10.The treatment group and model group mice intranasally infected with the virus solution 5ID₅₀,2h after infection were given oral administration of high,medium and low doses of Depestia and ribavirin,sustained drug delivery 7 days to observe the 14 days,animals were observed and recorded the reaction time of death and mortality.Influenza virus due to the impact of pneumonia:use the same method of infection and administration of sub-seven days,the animals were killed,sterile lung,said the lung weight,the naked eye to observe changes in the lungs,pulmonary index and calculation of inhibition rates of lung disease,and to do mouse lung tissue pathology observed.4.Homogenate of mouse lung tissue to do RT-PCR detection of influenza virus nucleic acid:Lung tissue after taking 100mg of total RNA extraction,reverse transcription reaction,real-time fluorescence quantitative PCR detection of influenza virus relative content of RNA,compared with model group,treatment group differences.This reaction also includes the design of primers to establish the reaction system,reaction conditions,such as steps to set up.Result:1.MTT assay:Concentration of the drug group the value of optical density OD (on behalf of the number of living cells) and virus control group,the optical density for inter-group t test,at high concentration,P <0.05,significant difference there is that agents can inhibit influenza proliferation of virus so that more cells survive.2.conventional detection,Depestia the dose groups of mice lung index virus were significantly lower than the model group,P <0.01;survival was significantly higher than the number of days virus model group P <0.05 or P <0.01.3.RT-PCR test:Influenza virus RNA levels(10⁵ copies / L) for the high dose group 39.29±6.06,in the dose group 36.09±8.19,low-dose group and 40.36±8.62 virus model group were 50.16±6.73 were significantly different,P<0.05.Conclusion:Depestia two sets of data in vitro therapeutic index(11.06 and 11.75),the Depestia to the treatment and prevention of influenza virus have shown that certain effects can be used as an anti-influenza virus of Chinese herbal medicine.Depestia in vivo lung index,the number of days survival,lung tissue content of influenza virus,lung tissue biopsy,etc.The results are that Depestia can inhibit or even kill the influenza virus,which weakened influenza virus infection on the lung tissue to improve influenza caused lung disease,with the role of anti-influenza virus.