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首页> 外文期刊>BMC Veterinary Research >Assessment of the hepatocyte protective effects of gypenoside and its phosphorylated derivative against DHAV-1 infection on duck embryonic hepatocytes
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Assessment of the hepatocyte protective effects of gypenoside and its phosphorylated derivative against DHAV-1 infection on duck embryonic hepatocytes

机译：人参皂甙及其磷酸化衍生物对鸭胚肝细胞的DHAV-1感染的肝细胞保护作用评估

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摘要

Duck viral hepatitis (DVH) is an acute disease of young ducklings with no effective veterinary drugs for treatment. Gynostemma pentaphyllum is a well-known traditional Chinese medicine that plays an important role in the treatment of various diseases. Gypenoside (GP), one of the main ingredients of Gynostemma pentaphyllum, was reported with good hepatoprotective effects. However, its low solubility limits its application in the clinics. To improve its solubility and bioactivity, a phosphorylated derivative of gypenoside (pGP) was prepared by the sodium trimetaphosphate-sodium tripolyphosphate (STMP-STPP) method. An infrared spectroscopy method was applied to analyse the structures of GP and pGP. Then, a methyl thiazolyl tetrazolium (MTT) colorimetric assay was applied to study the hepatocyte protective efficacy of these two drugs against duck hepatitis A virus type 1 (DHAV-1) infection, and qPCR, TUNEL labelling and flow cytometry methods were used to study the relevant hepatocyte protective in vitro. The infrared spectroscopy detection results showed that the phosphorylation modification of GP was successful. The MTT colorimetric assay results showed that both GP and pGP possessed good hepatocyte protective efficacy in vitro, and pGP performed better than GP when the drug was added before or after virus inoculation. Furthermore, the qPCR results revealed that both drugs could effectively inhibit the adsorption (when adding GP and pGP pre-virus inoculation), replication and release of DHAV-1, and the viral inhibition rate of pGP was greater than that of GP. The subsequent TUNEL labelling and flow cytometry assays showed that both GP and pGP could significantly inhibit duck embryo hepatocyte apoptosis induced by DHAV-1, and the inhibition effect of pGP was much stronger than that of GP. GP exerts good hepatocyte protective efficacy not only by inhibiting the proliferation of DHAV-1 but also by inhibiting duck embryonic hepatocyte apoptosis induced by DHAV-1, and phosphorylation modification significantly improves the antiviral and the anti-apoptotic effects of GP. Therefore, pGP has the potential to be developed into a novel drug against DHAV-1 infection.
机译：鸭病毒性肝炎（DVH）是一种雏鸭的急性疾病，没有有效的兽药治疗。绞股蓝（Gynostemma pentaphyllum）是一种著名的中药，在多种疾病的治疗中起着重要的作用。绞股蓝总皂甙的主要成分之一绞股蓝皂甙（GP）据报道具有良好的保肝作用。但是，其低溶解度限制了其在临床中的应用。为了提高其溶解度和生物活性，通过三偏磷酸钠-三聚磷酸钠（STMP-STPP）方法制备了人参皂甙（pGP）的磷酸化衍生物。采用红外光谱法分析了GP和pGP的结构。然后，采用甲基噻唑基四唑（MTT）比色法研究了这两种药物对鸭A型肝炎病毒（DHAV-1）感染的肝细胞保护作用，并使用qPCR，TUNEL标记和流式细胞术研究有关肝细胞的体外保护作用。红外光谱检测结果表明，GP的磷酸化修饰成功。 MTT比色法检测结果表明，GP和pGP在体外均具有良好的肝细胞保护作用，在病毒接种之前或之后添加GP时，pGP的性能均优于GP。进一步的qPCR结果表明，两种药物均能有效抑制DHAV-1的吸附（添加GP和pGP前病毒接种），复制和释放，且pGP的病毒抑制率高于GP。随后的TUNEL标记和流式细胞术分析表明，GP和pGP均能显着抑制DHAV-1诱导的鸭胚肝细胞凋亡，而pGP的抑制作用要强于GP。 GP不仅通过抑制DHAV-1的增殖，而且通过抑制DHAV-1诱导的鸭胚胎肝细胞凋亡而发挥良好的肝细胞保护作用，磷酸化修饰显着提高了GP的抗病毒和抗凋亡作用。因此，pGP有潜力被开发成针对DHAV-1感染的新型药物。

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