College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, PR China.
Poult Sci. 2019 May 1;98(5):1993-1999. doi: 10.3382/ps/pey557.
In this study, we examined the dose-dependent effects of the formula on Newcastle disease virus (NDV). In in-vitro test, the formula within safety concentration scope and NDV were added into cultured chick embryo fibroblast in 3 modes, and the cellular A570 values were determined by MTT (3-(4, 5-dimethyithiazol-2-yl)-2, 5-diphenyltetrazolium bromide) method. In in-vivo test, we examined the expression of interferon-induced transmembrane protein 3 (IFITM3) and Interferons (IFNs) in NDV-infected chickens. The results showed that the highest virus inhibitory rates of the formula at optimal concentration group were the highest (15.625 mg/mL) in post-adding and simultaneous-adding drug and virus modes, whereas medium concentration (7.813 mg/mL) showed the highest virus inhibitory rates in pre-adding drug mode. In vivo, the formula significantly upregulated the expression of IFITM3 in NDV-infected chickens at 3-D post-infection. However, the levels of IFNs were significantly downregulated. On days 5 and 7 post-infection, the levels of IFNs quickly upregulated. Moreover, the formula can significantly upregulate the antibody to resist the NDV compared with model control group on days 5 and 7 post-infection. In animals treated with the formula, the survival rate was nearly 37% higher at 7 d post-infection. We also found that the formula had a significantly stronger effect than a single herb on upregulating the expression of IFITM3. It confirmed that the formula could significantly inhibit the infectivity of NDV to chick embryo fibroblast. Also, the formula could significantly upregulated IFITM3 expression and inhibited virus replication in NDV-infected chickens. During the early stage of infection, IFNs were consumed to stimulate IFITM3 to inhibit virus replication, whereas during later stages of the infection, the formula upregulated the levels of IFNs and their antibodies to maintain a high level of immunity.
在这项研究中,我们研究了该配方对新城疫病毒(NDV)的剂量依赖性作用。在体外试验中,我们将在安全浓度范围内的配方和 NDV 以 3 种方式添加到培养的鸡胚成纤维细胞中,并通过 MTT(3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴化物)法测定细胞 A570 值。在体内试验中,我们检测了 NDV 感染鸡中干扰素诱导跨膜蛋白 3(IFITM3)和干扰素(IFNs)的表达。结果表明,在最佳浓度组中,该配方在加药后和同时加药及病毒模式下的最高病毒抑制率最高(15.625mg/ml),而中浓度(7.813mg/ml)在加药前模式下显示出最高的病毒抑制率。在体内,该配方在 NDV 感染后 3 天显著上调了 IFITM3 的表达。然而,IFNs 的水平显著下调。感染后第 5 和第 7 天,IFNs 的水平迅速上调。此外,与模型对照组相比,该配方在感染后第 5 和第 7 天可显著上调抗 NDV 的抗体。在接受该配方治疗的动物中,感染后第 7 天的存活率提高了近 37%。我们还发现,该配方在上调 IFITM3 表达方面的效果明显强于单一草药。这证实了该配方能显著抑制 NDV 对鸡胚成纤维细胞的感染力。此外,该配方能显著上调 IFITM3 的表达,抑制 NDV 感染鸡体内的病毒复制。在感染的早期,IFNs 被消耗以刺激 IFITM3 抑制病毒复制,而在感染的后期,该配方上调了 IFNs 及其抗体的水平,以保持高水平的免疫力。