Comparison Of Anti-NDV And Anti-IBV Activities Of Seven Chm Extracts And Their Sulfated Products

More and more researches have discovered that sulfation modification can enhance the biological activities of Chinese herbal medicinal polysaccharides or make polysaccharides generate new activities.In this research seven Chinese herbal medicines, Isatis root, Siberian solomonseal rhizome, Astragalus membranaceus, garlic, Epimedium, Chinese angelica and Codonopsis pilosula, were sleeted as object. Seven Chinese herbal medicinal extracts were extracted by water decoction method and modified by chlorosulfonic acid-pyridine method, and their anti-NDV and anti-IBV activities were compared. The purpose of this research was to investigate the possibility that sulfation modification enhance the antiviral activity of Chinese herbal medicinal extracts and offer theoretical evidence for development of antiviral Chinese herbal pharmaceutics. The tests are divided into the three parts:Test I Preparation of 7 Chinese herbal medicinal extracts and their sulfated products The 7 Chinese herbal medicinal polysaccharides were extracted by water-decocting and alcohol precipitation method. The 7 Chinese herbal medicinal extracts were extracted by water-decocting method. The content of polysacchride were determined by phenol-sulfuric acid methods. The results showed that the yield rate of GLPS was the highest (26.4%) and IRPS was the lowest (14.47%) in the seven polysaccharides. The content of polysaccharide of GLPS was the highest (93.39%), EPS was the lowest (20.4%). In the seven extracts, the yield rate of CPE was the highest (45.8%), IRE was the lowest (13.53%). The content of polysaccharide of GLE was the highest (85.35%), EPE was the lowest (32.5%). The content of the extracts were higher than that of polysaccharides content, so extract was selected for further experiments study object.The 7 Chinese herbal medicinal extracts were determined by phenol-sulfuric acid method. The sulfated modification conditions were refered to our laboratory. The results showed mat the product yield rate of sEPE was the highest (392%), sCPE was the lowest (53.33%). The content of polysacchride of sGLE was the highest (32.39%), sPSE was the lowest (18.19%). The Sulfate degree of substitution of sGLE was the highest (4.059), sPSE was the lowest (2.555).Test II Effects of 7 Chinese herbal medicinal extracts and their sulfated products on cellular infectivity of NDV The safe concentration of 7 Chinese herbal medicinal extracts and their sulfated products on chicken embryo fibroblast (CEF) were determined firstly. Five concentrations within safe concentration scope of theirs were added into cultivating system of CEF by three models with Newcastle disease virus (NDV), pre-, post-adding and simultaneous adding drugs and NDV. The A570 of cells were determined by MTT method to evaluate the effects (the A570 value and virus inhibitory rate) of these drugs on the cells effected NDV. The results showed that the drugs’A570 were significantly higher than the virus control group (P<0.05) at the concentration of 1 to 5 and indicated that the drugs could inhibit NDV effecting cells; When pre-adding drugs, the virus inhibitory rate of drug groups was in the range of 85.61%(GLE)～1.54%(EPE); The virus inhibitory rate of sEPE and sCAE was significantly higher than those of EPE and CAE (P<0.05), respectively. When post-adding drugs, the virus inhibitory rate of drug groups was in the range of 95.32%(sEPE)～11.54%(sASE); The virus inhibitory rate of sEPE was higher than that of EPE. When simultaneous adding drugs and NDV, the virus inhibitory rate of drug groups was in the range of 95.53%(sIRE)～24.34%(EPE); The virus inhibitory rate of sCAE, sPSE and sASE was significantly higher than those of CAE, PSE and ASE. Those suggested that sulfation modification could enhance anti-NDV activity of EPE, CAE, PSE and ASE.Test Ⅲ Effects of 7 Chinese herbal medicinal extracts and their sulfated products on cellular infectivity of IBV 7 Chinese herbal medicinal extracts and their sulfated products at five concentrations within safe concentration (including safe concentration) were added into cultivating system of CEF by three models with Infectious bronchitis virus (IBV), pre-, post-adding and simultaneous adding drugs and IBV. The A570 of cells were determined by MTT method to evaluate the effects (the A570 value and virus inhibitory rate) of these drugs on the cells effected IBV. The results showed that the drugs’ A570 were significantly higher than the virus control group (P<0.05) at the concentration of 1 to 5 and indicated that the drugs could inhibit IBV effecting cells; When pre-adding drugs, the virus inhibitory rate of drug groups was in the range of 95.90%(sGLE)～31.89%(IRE); The virus inhibitory rate of sCPE and sIRE was significantly higher than those of CPE and IRE (P<0.05), respectively. When post-adding drugs, the virus inhibitory rate of drug groups was in the range of 96.13%(GLE)-11.75%(PSE); The virus inhibitory rate of sPSE and sASE was significantly higher than that of PSE and ASE, respectively. When simultaneous adding drugs and IB V, the virus inhibitory rate of drug groups was in the range of 94.84%(sPSE)-38.57%(sGLE); The virus inhibitory rate of sEPE and sASE was significantly higher than those of EPE and ASE. Those suggested that sulfation modification could enhance anti-IB V activity of CPE s IRE, PSE, ASE and EPE.