摘要
目的研究丹参酮ⅡA磺酸钠(sodium tanshinoneⅡA sulfonate,STS)体外对呼吸道合胞病毒(respiratory syncytial virus,RSV)复制的影响及其机制。方法用CCK8法筛选对A549细胞不具细胞毒性的STS质量浓度,将RSV接种于A549细胞,2 h后加入STS,观察感染48 h后细胞病变效应(cytopathic effect,CPE),比较细胞培养上清中的病毒滴度。采用Vero细胞建立RSV感染模型,选取最佳效应质量浓度的STS处理后,观察CPE及测定上清中的病毒滴度。进一步在RSV感染A549细胞后的不同时间点给予STS处理及STS作用不同时间后去除STS,培养48 h后测定上清中病毒滴度。结果STS质量浓度<50.0μg/mL时,细胞生长不受影响,但当STS质量浓度升至100.0μg/mL时,A549细胞存活率明显下降。在RSV感染的A549细胞模型中,加入12.5、25.0、50.0μg/mL的STS处理48 h后均明显增加CPE的形成,以25.0 g/mL组效应最明显。加入12.5、25.0、50.0μg/mL的STS处理后,感染48 h时上清中病毒滴度均明显高于对照组,其中以25.0μg/mL的STS处理组病毒滴度最高,达4.88×10^(8) PFU/mL。在RSV感染的Vero细胞模型中,加入25.0μg/mL STS处理后,感染60 h时形成的CPE更大,范围更广泛,且病毒滴度明显高于对照组,达3.63×10^(8) PFU/mL,而RSV组上清中病毒滴度为5.88×106 PFU/mL。于感染RSV 0~24 h加入STS处理均明显增加病毒滴度,STS作用不同时间(6~24 h)后去除STS均不能增加病毒滴度。结论STS体外可促进RSV复制,不影响RSV入胞,在RSV感染的后期阶段促进病毒复制。本研究为进一步研究STS的药理学作用及为临床合理使用STS治疗RSV感染相关疾病提供一定的理论依据。
Objective To investigate the effect and mechanism of Sodium tanshinoneⅡA sulfonate(STS)on the replication of Respiratory syncytial virus(RSV)in vitro.Methods The CCK8 assay was used for screening on cytotoxic STS concentrations in A549 cells.RSV was inoculated into A549 cells,and then STS was added 2 h later.The cytopathic effect(CPE)was observed and the virus titers in the supernatant were compared at 48 h post-infection.Vero cells were infected with RSV and treated with the optimal concentration of STS,the CPE was observed and the virus titer in the supernatant was measured.Then STS was given at different time points(0-24 h)of RSV infection,and STS was removed from infected cultures at various times(6-24 h)post-infection.The virus titer in supernatant was determined at 48 h post post-infection.Results When the concentration of STS was less than 50.0μg/mL,the growth of A549 cells was not affected,but when the concentration of STS increased to 100.0μg/mL,the survival rate of A549 cells decreased significantly.In the RSV infected A549 cell model,the formation of CPE was significantly increased in the 12.5,25.0,50.0μg/mL STS treatment groups at 48 h after infection,and the 25.0 g/mL group had the most obvious effect.The virus titer in the supernatant of RSV infection at 48 h was significantly increased by 12.5,25.0,50.0μg/mL STS treatment,and the virus titer in the 25.0μg/mL STS treatment group was the highest(4.88×10^(8) PFU/mL).In the RSV infected Vero cell model,25.0μg/mL STS treatment significantly increased the CPE formed at 60 h post-infection,and the virus titer was significantly higher than that of the control group(3.63×10^(8) PFU/mL),while the virus titer in the supernatant of the RSV group was 5.88×10~6 PFU/mL.Treatment with STS at 0-24 h post-infection was all significantly promoted virus replication,however,the removal of STS at different time(6-24 h)could not promote virus replication.Conclusion STS promoted the replication of RSV in different cells in vitro.STS did not affect the entry of
作者
阳小凤
余福勋
金鑫
张梅
饶忠美
叶芝旭
YANG Xiaofeng;YU Fuxun;JIN Xin;ZHANG Mei;RAO Zhongmei;YE Zhixu(Department of Pediatrics,First Clinical Medical College of Zunyi Medical University;不详)
出处
《微生物学免疫学进展》
CAS
2023年第6期8-14,共7页
Progress In Microbiology and Immunology
基金
国家自然科学基金地区基金(81860003、81960001)
贵州省高层次创新型人才(千层次)(GZSYQCC-2023011号)
贵阳市科技局重大专项计划(筑科合同[2022]-4-1号)
贵州省中医药管理局中医药、民族医药科学技术研究课题(QZYY-2018-010)
贵州省卫生计生委科学技术基金项目(gzwjw2018-1-049)。
