山药多糖对PK-15细胞抗细小病毒感染能力的影响

Effects of Chinese Yam Polysaccharide on PK-15 cellular infectivity by PPV

采取一步醇沉和分步醇沉法提取4种山药多糖CYPS总、CYPS40、CYPS60、CYPS80.为研究山药多糖体外抗PPV活性,首先用MTT法测定山药多糖对PK-15的安全浓度;然后分别从125 ug/mL倍比稀释5个浓度,将4种山药多糖以先加山药多糖再加病毒的方式加至PK-15培养体系中,用MTT法检测山药多糖对PPV感染PK-15的影响;实时荧光PCR方法检测PK-15培养体系中PPV含量,观察山药多糖对PK-15清除PPV能力的影响.结果显示,在7.8～125 ug/mL范围内山药多糖组A570值和PPV含量分别显著高于和低于对照组,表明山药多糖可以增强PK-15抵抗PPV感染和清除PPV能力.山药多糖的这种作用效果与其浓度和醇沉浓度有关,山药多糖浓度越高则效果越好,分子量段以CYPS80的增强细胞活性和抗PPV感染效果最好.

Four kinds of Chinese Yam polysaccharides(CYPS),CYPSt,CYPS40,CYPS60and CYPS80,were extracted by one-step and stepwise ethanol precipitation methods.In order to going over the anti-PPV activity,first,determined the maximal safe concentration of CYPS by method of MTT(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide).And then four CYPS was diluted to five concentrations and added into PK-15monolayers by pre-adding drug pattern,to observe the antiviral activity of CYPS against PPV on PK-15by MTT assay.The concentration of PPV in foster system of PK-15was determined by fluorescence quantitative PCR assay to determine the effect of eliminating PPV of CYPS on PK-15.The results showed that CYPS could promote the anti-PPV ability of PK-15and reduce the content of PPV in foster system of PK-15in concentration of 7.8~125μg·mL-1.And this effect had related to concentration and ethanol precipitation concentration,that was the effectiveness better when concentration higher and the anti-PPV activity of CYPS80was best.