The Research On Extracts Of Commelina Antiviral Activity

Objective:Using the current more mature and easy percolation of technology and ion exchange techniques in Commelina total saponin extraction, so that these active ingredients to the greatest degree of retention, Then Commelina extract in vivo and in vitro anti-virus experiment, and, through its comparison with the positive drug efficacy and objective evaluation of Commelina extract anti-viral effect. Further evidence of development of Commelina antiviral value.Method:1. A-type H2N3 strains of influenza virus-induced mouse half of the deaths (LD50) Determination:The 10-fold serial dilution virus (10-1-10-8), each concentration of 4 mice inoculated (inhalants and abdominal injection to import), observed 2w, record mouse morbidity, mortality, calculate the LD50 value.2. Commelina extract on mice in vivo viral suppression test:A total of six experimental groups, respectively, Commelina extract three doses, namely, Tai (1.00g·kg-1·d-1), China (0.50g·kg-1·d-1) and small (0.25g·kg-1·d-1) dose. Ribavirin treatment group (0.24g·kg-1·d-1), viral infection group and the normal control group. Animal room to keep the air fresh and maintain the 60% relative humidity, temperature (20±4)℃, part of routine administration of disinfection, treatment group, viral infection group and the normal control group fed separately observed. Mice in each group with 10, Commelina communis extract treatment group were intraperitoneally injected, the virus dose used lethal dose (100LD50) 0.4ml/only by inhalation and intraperitoneal injection of type A,24h after administration, ribavirin group intraperitoneal injection of administration, all the continuous treatment group, administration 7d. The daily observation of animals, the response time of death and the deaths occurred. All animals observed after drug 14d. The body weight of mice in each group weighed and lung weight to calculate lung index, and lung tissue of virus hemagglutination test, the basic procedure is made of lung tissue homogenate inoculated chick embryo,2-4d collection of allantoic fluid, times the ratio diluted mixture were incubated with chicken red blood cell suspension, 1h after the observation of agglutination of red blood cells and calculate the virus HA titer.3. In vitro anti-influenza virus testing:Vero cell growth was single layer in 96-well plate, add 100TCID50 virus solution,0.1ml/hole,37℃,5% CO2 absorption 1h, abandoned to the virus solution, by adding TC50 (1.280mg/ml) 1/10 diluted 5-fold the concentration of pediatric fever paste extract to 4-fold serial dilution, that is, respectively,1280,320,80,20,5 ug/ml, and let the virus control (without liquid), cell control (without virus), subjects of drugs and positive drug control group (cells without viruses, plus corresponding to different concentrations of liquid). Repeat three times for each concentration,37℃,5% CO2 cultured 48h, observation of cytopathic effect, record pathological changes until the virus control cells in lesions up to 75% above the experimental end point, with the MTT assay the rate of viral suppression..4. Mouse lung tissue homogenate ELISA test done together of influenza virus detection: 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. (LD50) determination:mice infected with the virus, after a short incubation period (7d or so), will enter the disease state. After half of the titration of the virus-induced death of mice virulence (LD50) of 10-5.5/0.2ml..2. In vitro results showed that the extract of Commelina extract significantly inhibited in vitro the role of influenza virus, and has anti-influenza virus effect of ribavirin in comparison (40ug/ml), inhibit the role of influenza virus was no significant difference (P>0.05). Commelina extracts in experiments on mice with various doses of lung index were significantly lower than the virus model group, P<0.01; from the survival rate and the average survival time to read, each group showed significant differences among animals, and are known therapeutic effect than ribavirin, P>0.05. The normal control animals were not killed, the survival rate was 100%.3.The measured OD values compared with the virus was no significant difference (P>0.05). High-dose group and low dose group, significant difference P<0.01.Conclusion:Commelina experiment extracts significantly inhibited the role of influenza virus, which with the role of positive drug ribavirin compared with no significant difference (P>0.05), can determine Commelina extracts of influenza virus has a certain efficacy of prevention and treatment of role, and ribavirin equivalent can be used as an anti-influenza virus of Chinese herbal medicine. Immunotoxin Qing vivo lung index, survival days, the lung tissue concentration of influenza viruses, indicating Immunotoxin-ching indeed can inhibit or even kill the influenza virus, which weakened influenza virus infection on the lung tissue to improve the lung disease caused by influenza, with anti-influenza virus effect.