肺炎链球菌诱导SPLUNC1表达及白藜芦醇对SPLUNC1表达的影响

发布时间：2018-09-10 12:35

【摘要】：目的探讨肺炎链球菌(SP)感染时短腭、肺及鼻咽上皮克隆1(SPLUNC1)的宿主防御作用,及白藜芦醇(Res)对SPLUNC1表达的影响,为SP感染性疾病的治疗提供新的思路。方法根据感染复数(MOI)不同将SP感染BEAS-2B细胞分为对照组、MOI20 SP组和MOI50 SP组;根据Res药物浓度不同将Res预处理MOI20 SP感染BEAS-2B细胞分为12.5Res+SP组、25Res+SP组、50Res+SP组(Res终浓度分别为12.5μmol/L、25μmol/L、50μmol/L)。通过CCK-8法检测细胞活性,并选出SP、Res最佳作用浓度及时间。正式实验分为对照组、Res组、SP组和Res+SP组,采用实时定量PCR法和ELISA法检测各组SPLUNC1 m RNA及蛋白表达。结果随SP感染时间延长,细胞活性呈降低趋势;与对照组和MOI20 SP组相比,MOI50 SP组细胞活性有下降趋势;与MOI20 SP组相比,25Res+SP组细胞活性增强(P0.05),50Res+SP组细胞活性降低(P0.05)。以MOI20 SP菌悬液和25μmol/L Res进行正式实验。SP组和Res+SP组随感染时间延长,BEAS-2B细胞SPLUNC1m RNA水平表达先增高,后降低(P0.05);与SP组相比,各时间点Res+SP组SPLUNC1 m RNA及蛋白表达水平均增高(P0.05);与对照组相比,仅Res组SPLUNC1 m RNA及蛋白表达水平随时间延长未见明显改变(P0.05)。结论 SP感染可以诱导SPLUNC1表达,发挥宿主防御作用,Res可以在感染发生时上调SPLUNC1的表达,以浓度及时间依赖的方式加强细胞保护。
[Abstract]:Objective to investigate the host defense effect of short palate, lung and nasopharyngeal epithelial clone 1 (SPLUNC1) in Streptococcus pneumoniae (SP) infection and the effect of resveratrol (Res) on the expression of SPLUNC1 in order to provide a new idea for the treatment of SP infectious diseases. Methods BEAS-2B cells infected with SP were divided into control group and MOI50 SP group according to the complex number of (MOI) infection, and Res pretreated BEAS-2B cells with MOI20 SP infection were divided into 12.5Res SP group (Res 25 res SP group) (Res end concentration was 12.5 渭 mol/L,25 渭 mol/L,50 渭 mol/L) according to the different concentration of Res, and Res pretreated MOI20 SP infected BEAS-2B cells was divided into 12.5Res SP group with 25 res SP group (Res end concentration was 12.5 渭 mol/L,25 渭 mol/L,50 渭 mol/L). The cell activity was detected by CCK-8, and the optimum concentration and time of SP,Res were selected. The rats were divided into two groups: control group, res group, SP group and Res SP group. The expression of SPLUNC1 m RNA and protein in each group were detected by real-time quantitative PCR and ELISA. Results with the prolongation of SP infection time, cell activity decreased; compared with control group and MOI20 SP group, cell activity decreased in moi 50 SP group, and increased in MOI20 SP group (P0.05) compared with MOI20 SP group (P0.05). The expression of SPLUNC1m RNA in MOI20 SP suspension and 25 渭 mol/L Res group was increased first and then decreased with the prolongation of infection time (P0.05), compared with SP group, the expression of SPLUNC1m RNA in Res SP group and SP group increased at first and then decreased (P0.05), compared with SP group, the expression of SPLUNC1m RNA in Res SP group increased at first and then decreased (P0.05). The expression of SPLUNC1 m RNA and protein in Res SP group was higher than that in control group (P0.05), but the expression level of SPLUNC1 m RNA and protein in Res group did not change with time (P0.05). Conclusion SP infection can induce the expression of SPLUNC1. Res can up-regulate the expression of SPLUNC1 at the time of infection and enhance the cell protection in a concentration-and time-dependent manner.
【作者单位】：温州医科大学附属第二医院育英儿童医院儿童呼吸科;

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