| Purpose:To evaluate the streamlined formula and classic foemula extractions of Lonicerae and Forsythiae Powder extracted by 30% alcohol extraction artworks. The finger printings of main constituents of Lonicerae and Forsythiae Powder were obtained by high efficiency liquid chromatography and were combined with the experiment of inhibit influenza virus in vitro directly and the treatment and protection of chick embryos infected by influenza virus. Discuss the treatment of main extractions of Lonicerae and Forsythiae Powder to mouse pneumonia induced by influenza virus.Combination the chromatographic peak groups and IV-inhibitory effects to compare the differences between streamlined formula and classic foemula of Lonicerae and Forsythiae Powder.Material and method:1. The optimized artwork of Lonicerae and Forsythiae Powder was obtained by orthogonal experiment and 30% alcohol was used to prepare the main constituents of Lonicerae and Forsythiae Powder. Index contents of quality control of the main constituents of Lonicerae and Forsythiae Powder were used as the evaluation index to sieve and optimize the extraction artworks.2. The classic Powder and nine herbs finger printings were established by the high efficiency liquid chromatography and analysis the distribution features and focuses of finger printing capyluses of Lonicerae and Forsythiae Powder. Simultaneous determination of five components and discuss the relationship of finger printing capyluses.3. Lonicerae and Forsythiae Powder streamlined and classic foemula extractions with different extraction methods inhibiting IV in vitro in different times directly were observed by hemagglutination test; study the treatment and prevention of Lonicerae and Forsythiae Powder extractions to IV infected chick embryos.4. The pneumonia of young KM mouse were induced by IVâ… through cavitas nasi inoculation, hemagglutination test was used to detect hemagglutinin tite of lung tissue combining with the basic condition , pathological section of lung tissue, morphology and lung index of mouse to assess the IV pneumonia model of mouse to observe the treatment of lonicerae and forsythiae Powder streamlined and classic foemula extractions to IV mouse pnermonia.Results:1. The dosage of extraction solvents and extraction times were the significant factors in the quantity of mastic through orthogonal experiment, extraction time was the secondary factor, finally, the optimized extraction artwork was A2B2C2, which was refluxing 2 times with 30% alcohol and the volume of solvent was 15times, 10times to the materials, extracted 1.5 and 1 hours, respectively.2. Finger printings of different extraction artworks of Lonicerae and Forsythiae Powder have 26 fingerprint capylus, which belong to 9 single herbs.Acorrding to the quantity of chromatographic peak groups, Fructus arctii,Flos Lonicerae and Forsythia Suspense were the main herbs in the formula;according to the ares of chromatographic peaks, Fructus arctii, Radix Glycyrrhizae, Flos Lonicerae and Spica Schizonepetae, which means they are the main components of IV-inhibitory effects and proved the reason of herb recombination of streamlined formula of Lonicerae and Forsythiae Powder.3. Streamlined and classic foemula of Lonicerae and Forsythiae Powder inhibit the activity of IVâ… FM1 in vitro during from 1h to 24h, and the maximum non-blood clotting concentrations could inhibit the proliferation of virus at different extent; the effectiveness of anti-virus of streamlined foemula of Lonicerae and Forsythiae was most significant during the 6h-12h, which could go to 32 times and going to taper after 18h; the effectiveness of anti-virus of classic foemula of Lonicerae and Forsythiae was most significant at 12h, which could go to 32 times.4. Streamlined and classic foemula of Lonicerae and Forsythiae Powder anti influenza infected chick embryos: prevention and treatment, there was significant varieties of hemagglutinin valence between each extraction and virus control group(P<0.05). The anti-virus prevention of streamlined and classic foemula of Lonicerae and Forsythiae Powder were no significant(P>0.05);and no significance existed between the streamlined and classic foemula of Lonicerae and Forsythiae Powder(P>0.05); The anti-virus treatment of streamlined and classic foemula of Lonicerae and Forsythiae Powder were no significant(P>0.05);and no significance existed between the streamlined and classic foemula of Lonicerae and Forsythiae Powder(P>0.05).5. Each treatment groups was significant in improving the basic condition and pathological change of lung, degrading the IV hemagglutinin tites and lung index, but no varieties compared with Ribovirin group(P>0.05).Conclusion:1. Lonicerae and Forsythiae Powder was extracted by 30% alcohol, which obtained most active components in the herbs, the dosage of extraction solvent and extraction times were the important factors and extraction time was the second. Simultaneous determination of five components in the classic Lonicerae and Forsythiae Powder was a simple and reasonably with well reproducibility.2. The finger printing of classic Lonicerae and Forsythiae Powder complex has 26 generous fingerprint capyluses belong to the single herbs in the formula; The relative retention time and peak area ratio of generous capyluses could being as one of the quality control of formula; The chromatographic peaks in the chromatograms of medical material, intermedium and praeparatum obtained at the same chromatographic condition was well separated with satisfactory dependability, which satisfy the specification of finger printing.3. Streamlined and classic foemula of Lonicerae and Forsythiae Powder inhibit the virus in vitro during from 1h to 24h, the effectiveness was different at different times with dependability of time-potency and quantity-potency.4. Streamlined and classic foemula of Lonicerae and Forsythiae Powder extractions could prevent and treat the IV infected chick embryos in different extent; there was no variety in prevention and treatment effect between the Lonicerae and Forsythiae Powder groups and Ribovirin group; there was also no variety in IV-inhibitoy effects between the Lonicerae and Forsythiae Powder groups.5. Streamlined and classic foemula of Lonicerae and Forsythiae Powder extractions could relieve the lung inflammation, inhibit the virus replication and lessen the lung pathological damage. |
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