夏桑菊颗粒体外抗Ⅰ型登革热病毒的作用

Inhibitory effects of xiasangju granules on dengue virus type Ⅰ in vitro

目的研究夏桑菊颗粒体外抗Ⅰ型登革热病毒(DENV-1)的作用效果。方法 MTT法测定夏桑菊颗粒和利巴韦林对C6/36细胞的最大无毒剂量以及DENV-1感染细胞病变效应;细胞免疫荧光法,以NS1蛋白为目标蛋白,测定DENV-1经不同稀释度的夏桑菊颗粒预处理后感染能力;荧光定量PCR法检测夏桑菊颗粒和利巴韦林对DENV-1拷贝数的影响,采用方差分析比较不同稀释度药物对病毒的影响。结果MTT结果显示夏桑菊颗粒和利巴韦林的最大无毒剂量分别为7. 81 mg/m L和3. 13 mg/m L;经过夏桑菊颗粒溶液处理后,感染病毒的细胞存活率增加(21. 9±4. 3)%和(15. 7±4. 1)%,同时,经过利巴韦林处理后,细胞存活率分别增加(39. 2±1. 9)%和(24. 2±3. 1)%,两者均能明显减弱病毒引起细胞病变的效应(P <0. 05);荧光实验结果显示夏桑菊颗粒可使病毒相对感染率下降(42. 1±7. 4)%和(24. 2±2. 5)%,利巴韦林可使病毒相对感染率下降(51. 1±2. 3)%和(49. 3±2. 9)%,两者对病毒的感染具有明显的抑制作用(P <0. 05); q PCR实验结果表明夏桑菊颗粒对病毒拷贝数的抑制率达到(42. 1±7. 4)%和(24. 2±2. 5)%,利巴韦林对病毒拷贝数抑制率达到(51. 1±2. 3)%和(39. 3±2. 9)%,两者对病毒的RNA复制产生了明显的抑制作用(P <0. 05)。结论夏桑菊颗粒通过体外预给药的方式会产生明显的抗DENV-1的作用,其作用效果与利巴韦林相近。

Objective To evaluate the antivirus effect of Xiasangju granules against dengue virus serotype 1( DENV-1) in vitro. Methods Ribavirin injection was served as a positive control. The maximal nontoxic dose of Xiasangju granules and Ribavirin injection on the C6/36 cells was measured by MTT and used in subsequent experiments.The cytopathic effect on type 1 dengue virus was detected by MTT. The antiviral effect of Xiasangju granules on DENV-1 were determined by MTT,cell immunofluorescence and q PCR. Variance analysis was used to compare the effects of different dilution drugs on the virus. Results The maximum nontoxic dose of Xiasangju granules and Ribavirin injection on the C6/36 cells was 7. 81 mg/m L and 3. 13 mg/m L,respectively. MTT results showed that cell viability had been increased by( 21. 9 ± 4. 3)% and( 15. 7 ± 4. 1)% with Xiasangju granule solutions,and by( 39. 2 ± 1. 9)% and( 24. 2 ±3. 1)% with Ribavirin injection. The cytopathic effect of DENV-1 was significantly inhibited by Xiasangju granule and Ribavirin injection( P < 0. 05). According to the cell immunofluorescence results,the relative infection rate of DENV-1 was reduced by( 42. 1 ± 7. 4)% and( 24. 2 ± 2. 5)% with Ribavirin injection at the concentration of 3. 13 mg/m L and1. 56 mg/m L,respectively;and by( 51. 1 ± 2. 3)% and( 49. 3 ± 2. 9)% with Xiasangju granule at the concentration of7. 81 mg/m L and 3. 91 mg/m L,respectively. The DENV-1 infection rate was significantly suppressed by Ribavirin injection and Xiasangju granule( P < 0. 05). The q PCR results also showed that Xiasangju granule and Ribavirin injection had significantly inhibitory effect on RNA replication of DENV-1. Conclusion Xiasangju granule has obvious antivirus activity in vitro,which has similar effect of Ribavirin injection.