摘要
【目的】探讨热毒宁对呼吸道合胞病毒(RSV)感染人支气管上皮细胞(NHBE)分泌胸腺基质淋巴细胞生成素(TSLP)的影响。【方法】用Hep-2细胞扩增病毒,并进行毒力鉴定。建立并鉴定RSV感染NHBE的体外细胞模型。第一部分实验分RSV感染组和正常细胞对照组,感染组以不同滴度分为1000、500、100、50、10TCID505个组,感染12h、48h、72h、96h、120h,酶联免疫吸附(ELISA)法检测各组TSLP水平。第二部分实验将热毒宁以不同给药方式干预RSV感染NHBE,分为预防方式给药组、直接灭活病毒组、治疗方式给药组,并设正常细胞对照和病毒对照组,给药12h、24h、48h、72h,ELISA法检测各组TSLP水平。【结果】实时荧光定量RT-PCR法鉴定细胞模型建立成功。1.RSV对NHBE分泌TSLP的影响:相同感染时间内(12h、24h、48h、72h、96h、120h),随着感染滴度增加,各感染组TSLP分泌水平均呈上升趋势,与正常细胞对照组比较均有统计学差异(P均<0.05)。相同感染滴度内,随着感染时间增加(24h、48h、72h、96h、120h),各感染组TSLP分泌水平均呈上升趋势,与12h组比较均有统计学差异(P均<0.05)。2.热毒宁对RSV感染NHBE分泌TSLP的影响:在12h、24h、48h、72h给药时间内,热毒宁、利巴韦林的不同(预防、直接灭活、治疗)给药方式组与病毒对照组比较,均下调TSLP水平(P<0.05),以预防给药方式明显。【结论】RSV能诱导NHBE分泌TSLP,热毒宁可抑制RSV引起的TSLP分泌增加,以预防方式给药明显,热毒宁可能在病毒感染的控制中起到重要的作用。
[Objective] To study effects of traditional Chinese medicine Reduning injection on the secretion of TSLP in human bronchial epithelial cell infected by RSV in vitro.[Method] Hep-2 cell lines were used to pack and generate RSV.The RSV infectious titer was determined by TCID50 technique.An NHBE model of RSV infection in vitro was established and identified.First experiment included RSV infection-NHBE group and normal NHBE group.RSV-infected NHBE group was divided into 5 group based on RSV titers（1000,500,100,50,and 10 TCID50） in which each group also divided to different incubation hours（12 h,24 h,48 h,72 h,96 h,and 120 h）.The concentrations of TSLP in the culture supernatants were determined by enzyme-linked immunosorbent assay（ELISA）.Second experiment concern the effect of Redunig treatment on TSLP secretion based on cell culture in vitro.According to the way of Reduning delivery,experiments were divided into 3 groups（including precaution,direct deactivation and therapy）,accompanying a negative（normal NHBE） and a positive（NHBE-RSV） control group in which each group divided into 5 different incubation period（12 h,24 h,48 h,and 72 h）.The concentration of TSLP in the culture supernatants were also determined by ELISA.[Result] The mRNA of RSV was detected in total RNA of NHBE infected with RSV by real-time RT-PCR method,which demonstrated that a model of RSV infection in vitro was established successfully.1.The concentration of TSLP in NHBE cells supernatants infected by RSV significantly increased,compared with non-RSV.In the same post-inoculation time（12 h,24 h,48 h,72 h,96 h,and 120 h）,TSLP level increased obviously with raised RSV titers（P 0.05）.According to the same RSV titer（10,50,100,500,and 1000 TCID50）,TSLP level increased statistically in series infection time,compared with 12 h group（P﹤0.05）.2.The TSLP level of NHBE-RSV-Reduning group and NHBE-RSV-Ribavirin group were evidently lower than NHBE-RSV group in all detected points（12 h,24 h,48 h,and 72 h） and in all drug delivery ways（P 0.05）,especially in precaution way.（P 0.05）.TSLP level of NHBE-RSV-Reduning group are depressed than NHBE-RSV-Ribavirin group by precaution（P 0.05）.[Conclusion] TSLP secretion induced by RSV infection obviously increased with raised RSV titer and infected duration.Reduning injection observably inhibits TSLP secretion in RSV-infected NHBE with a positive time-effect relation.The inhibitory effect of precaution group was most significant among three drug delivery ways.The Reduning injection has a notable role in treating the viral diseases.
出处
《中山大学学报（医学科学版）》
CAS
CSCD
北大核心
2011年第2期203-207,212,共6页
Journal of Sun Yat-Sen University:Medical Sciences
基金
广东省自然科学基金(9451008901003013)
广东省科学事业费计划项目(2010B031600228)
