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为了研究RNA干扰(RNAi)对Ⅰ型登革病毒(DENV-1)在白纹伊蚊C6/36细胞内复制的影响,本研究设计并合成针对I型登革病毒Pr M基因的小干扰RNA,以脂质体法转染入C6/36细胞后,用DENV-1感染已转染的细胞,观察细胞病变效应,MTT法检测细胞存活率,荧光定量RT-PCR检测登革病毒RNA含量。结果表明:转染siRNA的C6/36细胞在受登革病毒攻击7天后仍无明显细胞病变效应,细胞存活率比对照组提高2.26倍,细胞内登革病毒RNA拷贝数比对照组降低约97.54%。说明利用RNA干扰技术能有效抑制登革病毒核酸在C6/36细胞内复制,并对细胞具有一定保护作用,为登革热的防治提供了新的思路。
To investigate the effect of RNAi on the replication of dengue virus type 1 (DENV-1) in C6 / 36 cells, we designed and synthesized a small interfering RNA against the Pr M gene of dengue I , And transfected into C6 / 36 cells by lipofectamine. The transfected cells were infected with DENV-1 to observe the cytopathic effect. The cell viability was detected by MTT assay and the RNA content of dengue virus was detected by real-time RT-PCR. The results showed that C6 / 36 cells transfected with siRNA still had no significant cytopathic effect after 7 days of challenge with dengue virus, the cell survival rate was 2.26 times higher than that of the control group, and the number of intracellular dengue virus RNA copies was reduced by 97.54 %. This indicated that the RNA interference technique can effectively inhibit the replication of dengue virus nucleic acid in C6 / 36 cells and has certain protective effect on the cells, providing a new idea for the prevention and treatment of dengue fever.