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Effect of Astragalus Polysaccharide and Isatis Root Polysaccharide on Proliferation of Chick Embryo Fibroblast and Inhibition of Porcine Parvovirus in Vitro

Author: ZhongHua
Tutor: ZhaoBaoYu
School: Northwest University of Science and Technology
Course: Clinical Veterinary Medicine
Keywords: Astragalus Polysaccharide (APS) Isatis Root Polysaccharide (IRPS) Chick Embryo Fibroblast (CEF) Cell Proliferation Porcine Parvovirus (P PV)
CLC: R285
Type: Master's thesis
Year: 2009
Downloads: 109
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Abstract


Astragalus and Isatis root were adopted as antiviral drugs to study the mechanism and immunological enhancement. The experiment was devided into 4 section, the details were summarized as following.1. Exaction and Determination of APS and IRPS.The two polysaccharides were extracted by decocting+ethanol precipitating. The contents of two polysaccharides were tested by Vitriol-anthracene ketone. The net contents of APS and IRPS were 15.3 g and 11.3 g and the exactive rates were 85.36% and 85.43%, respectively.2. Determination of Safe Concentration of APS and IRPS ON CEFAPS and IRPS were diluted with MEM medium into series of concentrations and were added in culture plates in two ways, at the beginning of culture and after CEF monolayer formation. Their maximal safety concentrations to CEF were judged according to OD value and morphological variety of the cells. The results showed that the maximal safety concentrations of APS and IRPS were 1 500 mg/mL and 650 mg/mL respectively.3. Effects of APS and IRPS on CEF proliferatingAPS and IRPS were diluted into five different levels within safety concentration were added into culture plates in two ways described previously. The cells OD were analysed after 24, 36, 48, 60 and 72 hours cultivation for the former groups and for latter groups to judge the effects of APS and IRPS on cell proliferation by MTT method. The results showed that APS and IRPS added with CEF simultaneously in the former groups could stimulate cell proliferation. Cell proliferation was also promoted by APS and IRPS added after CEF monolayer formation. It seemed that there were dosage and time-effect relationships.4. Inhibitive effect of porcine parvovirus by APS and IRPS in vitroInhibitive effect of porcine parvovirus was analysed on pig kidney (PK-15) cell monolayer of antiviral drugs by utilizing the cytopthic effect (CPE) inhibitive experiment when APS and IRPS were used alone and association by equal volumes. It was showed that APS had no obvious inhibitive effect on PPV by single use, but the IRPS had obvious inhibitive effect. the MIC was 0.58 mg/mL, MSC was 0.28 mg/mL; The anti-PPV cutivities of 1:1 mixture of APS and IRPS were significantly higher than that of Radix lsatidis, their MIC was 0.29 mg/mL and their MSC is 0.070 mg/mL.

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