腺病毒介导的白介素-24转移对脂多糖诱导的大鼠肾小球系膜细胞凋亡和周期调节蛋白p21、p27及CyclinE的影响

发布时间：2018-04-16 06:50

本文选题：腺病毒 + IL-　；参考：《中国循证儿科杂志》2014年05期

【摘要】：目的探讨腺病毒介导的IL-24转移对脂多糖(LPS)诱导的大鼠肾小球系膜细胞(GMCs)和细胞周期调节蛋白p21、p27及Cyclin E的影响。方法采用10%FBS/DMEM培养293细胞,扩增腺病毒载体,GMCs培养4~6代细胞后用于分析。1观察IL-24对LPS诱导的GMCs凋亡影响:分为对照组、Ad.IL-24组、LPS组和LPS+Ad.IL-24组,其中对照组和LPS组GMCs不转染携带IL-24腺病毒(Ad.IL-24),Ad.IL-24组和LPS+Ad.IL-24组以Ad.IL-24转染GMCs,LPS组和LPS+Ad.IL-24组加LPS培养,以流式细胞术检测培养后24和48 h的GMCs凋亡率;2考察IL-24对GMCs周期蛋白影响:对照组为5%FBS/DMEM培养GMCs,Ad-GFP组采用携带绿色荧光蛋白的对照载体(Ad.GFP)转染GMCs,Ad.IL-24+LPS组以Ad.IL-24转染GMCs,采用Western-blot检测p21、p27及Cyclin E表达。结果 1GMCs在培养后24和48 h细胞凋亡率:对照组分别为(0.86±0.15)%和(0.98±0.4)%;IL-24组分别为(1.02±0.22)%和(1.43±0.31)%;LPS组分别为(2.19±0.81)%和(2.49±0.12)%,LPS+Ad.IL-24组分别为(18.01±1.17)%、(26.82±5.01)%;2个观察时间点细胞凋亡率对照组与IL-24组差异无统计学意义,LPS组显著高于对照组(P0.05),LPS+Ad.IL-24组细胞凋亡率最高(P0.05)。2Ad-GFP组p21、p27表达显著低于对照组(P0.05),Cyclin E表达显著高于对照组(P0.05);Ad.IL-24+LPS组p21、p27表达较Ad-GFP组显著上调(P0.05),Cyclin E表达较Ad-GFP组下调。结论 IL-24腺病毒载体对正常GMCs无影响,但可使LPS诱导增生的GMCs凋亡增加,其可能机制是上调关键细胞周期负调控蛋白p21、p27及下调Cyclin E。
[Abstract]:Objective to investigate the effects of adenovirus-mediated IL-24 transfer on rat glomerular Mesangial cells (GMC) and cell cycle regulatory proteins p21, p27 and Cyclin E induced by lipopolysaccharide (LPS).Methods 293 cells were cultured with 10 S / DMEM and cultured with adenovirus vector GMCs for 4 ~ 6 passages. The effects of IL-24 on GMCs apoptosis induced by LPS were analyzed. The cells were divided into two groups: control group, Ad.IL-24 group and LPS Ad.IL-24 group.The GMCs of control group and LPS group were not transfected with Ad.IL-24, Ad.IL-24, and LPS Ad.IL-24 group were transfected with Ad.IL-24 and LPS Ad.IL-24 group with LPS.The apoptosis rate of GMCs at 24 and 48 hours after culture was detected by flow cytometry. The effect of IL-24 on GMCs cyclin was investigated. The control group was cultured with 5 S / DMEM. The control group was transfected with GMCsN Ad-GFP vector Ad.GFP carrying green fluorescent protein (GFP), and the GMCsN + +. IL-24 LPS group was transfected with Ad.IL-24.The expression of p21 p27 and Cyclin E was detected by Western-blot.Results the apoptotic rates of 1GMCs were 0.86 卤0.15% and 0.98 卤0.4% in the control group and 1.02 卤0.22% and 1.43 卤0.31% in the control group, respectively, and 2.19 卤0.81% and 2.49 卤0.12% in the LPS group and 2.49 卤0.12% in the IL-24 group, respectively, and there was no significant difference between the control group and the IL-24 group at 2 time points.The expression of p21p27 in LPS group was significantly higher than that in control group (P0.05 Ad.IL-24). The expression of p21p27 was significantly lower in LPS group than that in control group P0.05P0.05. IL-24 LPS group was significantly higher than that in Ad-GFP group, and the expression of p21 p27 was significantly up-regulated in Ad-GFP group than in Ad-GFP group.Conclusion Adenoviral vector of IL-24 has no effect on normal GMCs, but it can increase the apoptosis of proliferative GMCs induced by LPS. The possible mechanism is to up-regulate the critical cell cycle negative regulatory protein p21p27 and down-regulate Cyclin E.
【作者单位】：大连医科大学附属第一医院儿科;华中科技大学同济医学院附属同济医院儿科;上海东方医院普外科;
【分类号】：R3416

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