| Radix Isatidis is one of the most important antiviral anti-inflammation Traditional Chinese Medicine. Some glycoproteins and polysaccharides, such as lectin, have been reported to be its antiviral principles, but lack of evidences to prove their effects in vivo. The mechanism studies of its therapeutic effects on influenza were focused on the virus itself. The cell protection effects of Radix Isatidis water Extract (RIE) have not been carried on in any publications.Recently, RIE was proved to inhibit the adsorption of influenza viruses to Red Blood Cells (RBC) and MDCK cells in vitro by Institute of Biotechnology,Fuzhou University. RIE can bind to cell membranes and modify the chemical and physical properties of cell membranes. This study launchs some methods like the in vivo antiviral test, MDCK cells cultivation and anti-oxidation activity determinations, in order to prove the cell protection and antiviral effects of RIE and illuminate its mechanism.Firstly, an in vivo test of RIE's effects on influenza viruses induced human RBC agglutination was done before and after RIE oral administration. RBC of type-0 blood from a healthy young male was collected and incubated with Type A influenza viruses (1:32) for 60mins. Agglutination was investigated by microscopy study( X 400). By contrast, RBC of 150mins after oral administration of RIE(150mg/Kg.bw) to the same volunteer was collected and incubated with viruses for 60mins. No agglutination was observed in microscopy studies. Furthermore, the RBC collected before RIE administration were incubated with serum collected 150mins after RIE administration for 60mins, and then were incubated with viruses for another 60mins. No agglutination was observed either. For the first time,these results approve that the cell protective components from RIE can be absorbed into blood and work in vivo. This effects of RIE is carried out by altering the components of serum and surface characterization of RBC.Secondly, some RIE components acted on influenza viruses directly in the MDCK cell cultivation. MDCK cells were divided into 3 groups: A, B and C. In group A, cells were incubated with RIE at gradient concentration of 2mg/ml, 1mg/ml,500μg/ml,250μg/ml,125μg/ml,62.5μg/ml for 120mins, respectively. And then viruses (titre=1:8) were inoculated in after RIE was removed. In group B, after 60 mins' incubation with RIE, viruses and RIE were incubated with MDCK cells together. In group C, cells were treated with RIE for 120mins and then inoculated viruses without removing RIE. After 3 days' cultivation, the virus titres of every group were determinated. As the results shown, viruses in all of the three groups were inhibited at vary degree. Their minimum inhibition concentration are 500μg/ml, 500μg/ml, 1mg/ml respectively. The titre of group B at 1mg/ml or upper concentrations was undetectable, which means the viruses proliferation was completely inhibited. This result indicates that the ingradients from RIE can have effects on viruses directly. To illustrate the mechanism of this action, the combinating ability of RIE to model proteins was investigated. Since BSA and CEA precipitated partly from their solutions after 1 hour inbubation with RIE at 37℃, the interaction between RIE and proteins may contribute to the inhibition of viruses.
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