本文選題：腺病毒 + IL-��； 參考：《中國循證兒科雜志》2014年05期

【摘要】：目的探討腺病毒介導(dǎo)的IL-24轉(zhuǎn)移對脂多糖(LPS)誘導(dǎo)的大鼠腎小球系膜細(xì)胞(GMCs)和細(xì)胞周期調(diào)節(jié)蛋白p21、p27及Cyclin E的影響。方法采用10%FBS/DMEM培養(yǎng)293細(xì)胞,擴增腺病毒載體,GMCs培養(yǎng)4~6代細(xì)胞后用于分析。1觀察IL-24對LPS誘導(dǎo)的GMCs凋亡影響:分為對照組、Ad.IL-24組、LPS組和LPS+Ad.IL-24組,其中對照組和LPS組GMCs不轉(zhuǎn)染攜帶IL-24腺病毒(Ad.IL-24),Ad.IL-24組和LPS+Ad.IL-24組以Ad.IL-24轉(zhuǎn)染GMCs,LPS組和LPS+Ad.IL-24組加LPS培養(yǎng),以流式細(xì)胞術(shù)檢測培養(yǎng)后24和48 h的GMCs凋亡率;2考察IL-24對GMCs周期蛋白影響:對照組為5%FBS/DMEM培養(yǎng)GMCs,Ad-GFP組采用攜帶綠色熒光蛋白的對照載體(Ad.GFP)轉(zhuǎn)染GMCs,Ad.IL-24+LPS組以Ad.IL-24轉(zhuǎn)染GMCs,采用Western-blot檢測p21、p27及Cyclin E表達(dá)。結(jié)果 1GMCs在培養(yǎng)后24和48 h細(xì)胞凋亡率:對照組分別為(0.86±0.15)%和(0.98±0.4)%;IL-24組分別為(1.02±0.22)%和(1.43±0.31)%;LPS組分別為(2.19±0.81)%和(2.49±0.12)%,LPS+Ad.IL-24組分別為(18.01±1.17)%、(26.82±5.01)%;2個觀察時間點細(xì)胞凋亡率對照組與IL-24組差異無統(tǒng)計學(xué)意義,LPS組顯著高于對照組(P0.05),LPS+Ad.IL-24組細(xì)胞凋亡率最高(P0.05)。2Ad-GFP組p21、p27表達(dá)顯著低于對照組(P0.05),Cyclin E表達(dá)顯著高于對照組(P0.05);Ad.IL-24+LPS組p21、p27表達(dá)較Ad-GFP組顯著上調(diào)(P0.05),Cyclin E表達(dá)較Ad-GFP組下調(diào)。結(jié)論 IL-24腺病毒載體對正常GMCs無影響,但可使LPS誘導(dǎo)增生的GMCs凋亡增加,其可能機制是上調(diào)關(guān)鍵細(xì)胞周期負(fù)調(diào)控蛋白p21、p27及下調(diào)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 鹵0.15% and 0.98 鹵0.4% in the control group and 1.02 鹵0.22% and 1.43 鹵0.31% in the control group, respectively, and 2.19 鹵0.81% and 2.49 鹵0.12% in the LPS group and 2.49 鹵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.
【作者單位】： 大連醫(yī)科大學(xué)附屬第一醫(yī)院兒科;華中科技大學(xué)同濟(jì)醫(yī)學(xué)院附屬同濟(jì)醫(yī)院兒科;上海東方醫(yī)院普外科;
【分類號】：R3416

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