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| In vitro Inhibition of Polygonum cillinerve Polysaccharide on Swine transmissible gastroenteritis virus (TGEV) |
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Abstract Swine transmissble gastroenteritis(TGE) is a kind of hyperinfected viral disease in pigs (Sus scrofa) induced by Swine transmissble gastroenteritis virus (TGEV), and leads to death of large numbers of the pigs. The present study aimed to evaluate the effect of Polygonum cillinerve polysaccharide (PCP) on inhibition of TGEV. Based on the detection results of TCID50 of TGEV and maximum cytotoxic concentration (TC0) of PCP on in vitro growth of swine testicular (ST) cells, the PCP was pro-seeded, co-seeded or post-seeded with TGEV in ST cells, then the anti-virus adsorption, inactivation and inhibition of the proliferation of PCP were analyzed by 3- (4,5- Dimethylthiazol-2- yl)-2,5- diphenyltetrazolium bromide (MTT) method and quantitative Real-time PCR (qRT-PCR). The detection of in vitro viral infection indicated that the value of TCID50 on TGEV was 10-6.25. After 72 h in vitro culture, ST cells were well-grown without the cytotoxic response when treated them with PCP at concentration of lower than 20 μg/mL compared with the blank controls, which determined the TC0 of PCP was 20 μg/mL. According to the results of MTT, under the maximum safe concentration (20 μg/mL) of PCP, the anti-adsorption effect of PCP on TGEV in vitro was the best, the inhibition rate was 85.2%, the inhibition rate on in vitro virus proliferation was 78.4%, while the inhibition rate of direct inactivation to TGEV was 74.4%, and the inhibition rate was positively correlated with the concentration. Moreover, qRT-PCR detection indicated that 3 administration methods of PCP could significantly reduce relative TGEV N mRNA expression at the transcriptional level (P＜0.01) compared with the virus control, and with a dose-dependent characteristic. The results suggested that all 3 administration methods of PCP could inhibit the proliferation of TGEV at ST cells in vitro, and the prevention effect of PCP was the best. This work demonstrated the in vitro anti-virus effect of PCP on TGE, and can provide some basis on resource exploitation and multiple utilization of Polygonum cillinerve.
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Received: 26 May 2014
Published: 13 January 2015
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