Observation On The Antivirus Function Of Total Flavone Ingredients And Their Prescriptions In Vitro

In order to screen better flavone prescriptions of anti-NDV, flavone prescriptions were formed by the flavones which showed better antivirus activities in vitro. This experiment was formed by the following parts:Experiment1Extraction and the maximal safe concentration of total flavones in traditional Chinese herbal medicines on CEF The total flavones of nine traditional Chinese herbal medicines, such as Sanchi (SF), Baikal Skullcap Root (BSR), Radix Puerariae (RP), Barbed Skullcap Herb (BSH), Japanese Honeysuckle Stem (JHS), Chinese Dodder Seed (CDS), drynaria (DF), Wild Dendranthema Flower (WDF) and Common Solidago Herb (CSH) were extracted by ethanol-reflux method with acid-alkali-precipitation method. The maximal safe concentrations of the ten total flavones on chick embryo fibroblast (CEF) were determined by MTT colorimetric assay (Epimedium total flavones (EF) were preserved by our laboratory). The results showed that the extractive rate of Baikal Skullcap Root total flavones (BSRF) was8.40%, which was the highest in the nine total flavones. However, Japanese Honeysuckle Stem total flavone (JHSF) was in contrast. The maximal safe concentrations of SF, BSRF, RPF, BSHF, CDSF, WDFF, CSHF, EF, JHSF, DF on CEF were31.3,62.5,62.5,62.5,62.5,62.5,62.5,62.5,125and125μg·mL-1respectively. Moreover, the ten total flavone ingredients foresaid within safe concentrations could promote cell proliferation and prolong cell survival time in some degree.Experiment2Effects of ten total flavone ingredients on NDV infectivity in CEF In order to screen flavone ingredients with better antiviral effect, ten total flavone ingredients were diluted with MEM medium into five series concentrations from the maximal safe concentration, then added into CEF monolayer in three ways:adding drug firstly, adding virus firstly and adding drug and virus simultaneously. MTT colorimetric assay was used to detect the effects of these flavones anti-Newcastle disease virus (NDV) in chick embryo fibroblast (CEF). The results indicated that all of the flavones showed antiviral activities in different degree. The A570values of EF at31.3-15.6μg·mL-1with the treatments of adding drug firstly and adding virus firstly respectively, and at31.3μg·mL-1when adding drug and virus simultaneously, and of BSRF at7.8-3.9μg·mL-1when adding virus firstly and WDFF at62.5-31.3μg·mL-1when adding drug firstly were significantly higher than those of cell control and virus control (P<0.05). It suggested these flavones with better antiviral activities. Considering the antivirus abilities treated by the3above methods and the extraction ratio, four total flavones of EF, BSRF, WDFF and SF were choosed as the preliminary flavone ingredients of the flavone ingredient prescriptions of antivirus.Experiment3Effects of Chinese herbal total flavone ingredient prescriptions on NDV infectivity in CEF In order to screen better flavone prescriptions of antivirus in vitro, nine total flavone ingredients prescriptions were formed based on the four total flavones of EF, BSRF, WDFF, and SF, which showed better antivirus activities described in last chapter. Every prescription was diluted into five series concentrations from its maximal safe concentration, then, added into CEF monolayer with three ways. MTT colorimetric assay was used to detect the effects of the nine prescriptions anti-NDV in CEF. The results indicated that all of the prescriptions displayed antivirus abilities in some degree when treated by the above three treatments. Evaluated by A570values and the highest virus inhibition ratio, three prescriptions of Epimedium total flavones-Baikal Skullcap Root total flavones (EF-BSRF), Epimedium total flavones-Wild Dendranthema Flower total flavones-Baikal Skullcap Root total flavones (EF-WDFF-BSRE), Epimedium total flavones-Sanchi total flavones-Wild Dendranthema Flower total flavones-Baikal Skullcap Root total flavones (EF-SF-WDFF-BSRF) showed better antivirus activities, then followed by the prescription of Epimedium total flavones-Wild Dendranthema Flower total flavones-Sanchi total flavones (EF-WDFF-SF).