| Objective: To observe anti-viral effects of Qing Re Mixture(QRM.)onrespiratory syncyntial virus (RSV), adenovirus type 3 (adv3), influenza virustype A3, A1, B and study on active constituent of QRM.absorbed into plasmaby establishing a RP-HPLC method for determination of baicalin in plasmaso as to provide scientific basis for its development as a new drug. Methods: (1) In vitro anti-virus activity by means of cell culture techniquesThe anti-respiratory syncyntial virus(RSV) , anti-adenovirus type 3 (adv3)and anti-influenza virus type A3, A1, B effects of QRM.were respectivelyobserved by means of the technique of Vero, Hep-2 cell and MDCK cellculture using Shuang Huang Lian as a positive control. (2) In vitro anti-virusactivity by means of chick embryo culture techniques The techniques ofchick embryo culture were used for evaluating antiviral activity of serumfrom the Wistar rats dosed with QRM.by hemagglutination assay with theinfluenza virus type A3 hemagglutination titer as index. (3) Dynamicchange of baicalin of QRM.in serum of rats Rat's plasma was pretreatedby deproteinization with methanol, evaporation of the supernatant to dryunder the nitrogen steam and dissolution of the residue with small amountof the mobile phase. The chromatographic sepatation of baicalin wasachieved on a C18 column. Baicalin was eluted with mobile phaseconsisting of methanol and pH3.5 phosphate buffer (50:50, V/V). Results: (1) In vitro anti-RSV activity by means of cell culture techniques InVero and Hep-2 cell culture, QRM.was a potential inhibitor of RSV inconcentration-dependent manner(P<0.01). In Vero cell culture the TD50and IC50 were 31.62mg·mL-1 and 0.50mg·mL-1, respectively, thus TI wascalculated 63.24. In Hep-2 cell culture the TD50 and IC50 were32.10mg·mL-1 and 0.54mg·mL-1, respectively, thus TI was calculated 59.44.(2) In vitro anti-adv3 activity by means of cell culture techniques In Veroand Hep-2 cell culture, QRM . was a potential inhibitor of adv3 inconcentration-dependent manner(P<0.01). In Vero cell culture the TD50and IC50 were 31.62mg·mL-1 and 0.45mg·mL-1, respectively, thus TI wascalculated 70.27. In Hep-2 cell culture the TD50 and IC50 were32.10mg·mL-1 and 0.52mg·mL-1, respectively, thus TI was calculated 61.73.(3) In vitro anti-influenza virus type A3, A1, B activity by means of cellculture techniques In MDCK cell culture, the TD50 of QRM.was 28.04mg·mL-1, TD0 hasn't anti-influenza virus type A3, A1 effect. The IC50 was5.20 mg·mL-1 and TI was 5.39 during study on anti-influenza virus type B.(4) In vitro anti-influenza virus type A3 activity by means of chick embryoculture techniques Serum taken at 1, 2, 3, 4 h post dosing to the Wistarrats at a dose of 24g·kg-1 by oral route showed antiviral effect withinhibitory rates (%) of hemagglutination titer were 86.37, 82.32, 80.72 and72.74, respectively. Serum taken at 1, 2, 3 h post dosing to the Wistar rats ata dose of 12 g·kg-1 by oral route showed antiviral effect with inhibitoryrates (%) of hemagglutination titer were 71.53, 67.58 and 66.14,respectively. QRM.was found to be an inhibitor of influenza A3 virus in aconcentration-dependent manner. (5) Dynamic change of baicalin ofQRM.in serum of rats The linear range of baicalin was 0.025 ~2.5μg·mL-1 and the lower limit of quantitation (LLOQ) is 0.025 μg·mL-1. Theaccuracy and precision meet the requirement of biological specimenanalysis. The results of animal experiment showed that the baicalin inQRM.presented multiple-peak phenomenon in the body of Wistar rats. Conclusion: QRM.has a significant anti-RSV, anti-adv3 effect in Vero and Hep-2cell culture; QRM.has anti-influenza virus type B effect and hasn'tanti-influenza virus type A3,A1 effect in MDCK cell culture; QRM.hassignificant effect on influenza virus type A3 in the chick embryo; RP-HPLCmethod developed by us is simple, rapid and suitable for the determinationof plasma concentration of baicalin and the study of pharmacokinetics.Thebaicalin in QRM.presented multiple-peak phenomenon in the body ofWistar rats and was proved to be possib...
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