桃金娘鞣质抗猪流行性腹泻病毒作用研究

为测试桃金娘鞣质对猪流行性腹泻病毒(PEDV)的影响,体外试验以PEDV感染PK细胞为模型,采用细胞病变(CPE)效应法、MTT比色法检测细胞存活率,测定桃金娘鞣质抑制PEDV增殖的半数有效浓度(EC_(50))和治疗指数(TI),同时设利巴韦林对照组;体内试验以PEDV感染昆明种小鼠为模型,60只昆明种小鼠随机均分为6组,设桃金娘鞣质高剂量组、中剂量组、低剂量组、利巴韦林对照组、病毒对照组、健康对照组,采用实时荧光定量PCR法检测肠系膜淋巴结PEDV含量,观察桃金娘鞣质抗PEDV增殖能力。结果显示,桃金娘鞣质能明显抑制PEDV引起的细胞病变,半数有效浓度为0.021 mg/mL,不如利巴韦林0.0098 mg/mL(P0.05);治疗指数为361.9,优于利巴韦林214.28(P0.05);桃金娘鞣质能影响PEDV在小鼠体内的增殖数量,并呈现一定量效关系;桃金娘鞣质各剂量组小鼠肠系膜淋巴结PEDV含量均低于病毒对照组(P0.05),高、中剂量组小鼠肠系膜淋巴结PEDV含量均低于利巴韦林对照组(P0.05),小鼠肠系膜淋巴结PEDV含量在桃金娘鞣质高、中、低剂量组间也存在显著差异(P0.05)。试验表明,桃金娘鞣质具有体外、体内抗PEDV药理活性,有进一步开发抗病毒药物的价值。

Study on the Effects of Myrtle Tannins on Porcine Epidemic Diarrhea Virus

The experiment was aimed to study the effects of Myrtle tannins on porcine epidemic diarrhea virus. In vitro experiment, porcine epidemic diarrhea virus was used to infect competitive cells as a model. The cell survival rate was observed by cytopathic effect (CPE) method and MTT colorimetric method. The half effective concentration (EC50) and therapeutic index (TI) of Myrtle tannins in inhibiting the proliferation of PEDV were determined and ribavirin was set up as control group at the same time. In vivo experiment, Kunming mice infected with PEDV were used as the model. Sixty Kunming mice were randomly divided into 6 groups, namely, the normal control group, the ribavirin group, the virus control group, the high-dose myrtle tannins group, the middle-dose group, and the low-dose group. PEDV content in mesenteric lymph nodes was detected by real-time fluorescence quantitative PCR to evaluate the anti-PEDV proliferation ability of myrtle tannins. In vitro experiment results showed that Myrtle tannins can effectively inhibit cytopathic effect induced by PEDV. The EC50 was 0.021 mg/mL, which was lower than that of ribavirin 0.0098 mg/mL (P<0.05), and the TI was 361.9, which was better than that of ribavirin 214.28 (P<0.05). In vivo experiment results showed that Myrtle tannins could affect the proliferation of PEDV in mice, and showed a certain dose-effect relationship. The content of PEDV in mesenteric lymph nodes of mice in each Myrtle tannins group was lower than that in the virus control group (P<0.05) and the content of PEDV in mesenteric lymph nodes in high and middle tannins groups was lower than that in ribavirin group (P<0.05). There was also a significant difference in PEDV content in mesenteric lymph nodes among high, middle and low tannins groups (P<0.05). In conclusion, Myrtle tannins had anti-PEDV pharmacological activities in vitro and in vivo, which was value of further development of antiviral drugs.