槲皮素对人胚肺成纤维细胞分化过程中PI3K/Akt信号通路影响  

Effect of quercetin on the expression of PI3K/Akt signaling pathway in human embryonic lung fibroblasts differentiation

作　　者：彭海兵[1] 刘燕[2] 田景瑞[2] 穆玉[1] 王建行[1] 王献华[2] 

机构地区：[1]河北联合大学冀唐学院,河北唐山06300 [2]河北联合大学基础医学院,063000

出　　处：《中国职业医学》2014年第3期286-289,共4页China Occupational Medicine

基　　金：唐山市科技局科研基金(13130296z)

摘　　要：目的探讨槲皮素对人胚肺成纤维细胞(HELF)分化过程中磷脂酰肌醇3-激酶(PI3K)信号通路的影响。方法支气管肺泡灌洗法收集矽肺患者肺泡巨噬细胞(AM),离体用含有二氧化硅(SiO2)粉尘(质量浓度为50mg/L)的达尔伯克氏改良伊格尔(DMEM)培养基和不含SiO2的DMEM培养基分别培养18 h,收集AM上清液,在含SiO2的上清液中分别加入终浓度为10、20、40μmol/L的槲皮素。用组织贴块法获取培养HELF,分为空白对照组、AM组、(SiO2+AM)组、槲皮素低浓度组、槲皮素中浓度组和槲皮素高浓度组。蛋白印迹法检测HELF中磷酸化的蛋白激酶B(p-Akt)的表达,免疫细胞化学法检测HELF中α-平滑肌肌动蛋白(α-SMA)的表达。结果除槲皮素高浓度组外,其余各处理组p-Akt表达水平分别高于空白对照组(P<0.05)。(SiO2+AM)组p-Akt表达水平高于AM组(P<0.05);3个浓度的槲皮素组p-Akt表达水平分别低于AM组和(SiO2+AM)组(P<0.05);槲皮素高浓度组p-Akt表达水平分别低于槲皮素中、低浓度组(P<0.05)。除槲皮素高浓度组外,其余各处理组α-SMA表达水平均高于空白对照组(P<0.05);(SiO2+AM)组α-SMA表达水平高于AM组(P<0.05);除槲皮素低浓度组外,槲皮素中、高浓度组α-SMA表达水平分别低于AM组(P<0.05)。3个浓度的槲皮素组α-SMA表达水平分别低于(SiO2+AM)组(P<0.05)。槲皮素高浓度组α-SMA表达水平分别低于槲皮素中、低浓度组(P<0.05)。结论槲皮素可以抑制HELF分化过程中PI3K/Akt信号通路,从而下调α-SMA的表达。Objective To explore the effect of quercetin on the expression of phosphatidylinositol 3-kinase signaling path-way in human embryonic lung fibroblast （HELF） differentiation.Methods The silicotic alveolar macrophages （AM） were collected by bronchoalveolar lavage and then incubated in vitro with DMEM medium with or without SiO 2 （50 mg/L） for 18 hours.Then the AM supernatant incubated for 18 hours was collected , and in the supernatant containing SiO 2 was added with quercetins as the stimulus supernatant .HELF was isolated by organize paste block method .Four generations of pul-monary fibroblasts were divided into control group , AM group, （SiO2 ＋AM） group and quercetin in low, middle, high concentration （10, 20, 40μmol/L respectively ） group.The expression of phosphorylation of protein kinase B （ p-Akt） in the HELF was measured by western blot and α-SMA in the HELF was measured by immunocytochemistry method .Results Except for the quercetin in high concentration group , the p-Akt expression levels in other four groups were higher than that of the control group（P〈0.05）.The （SiO2 ＋AM） group was higher than the AM group （P〈0.05）.The quercetin groups were lower than those of the AM group and （SiO2 ＋AM） group （P〈0.05）.Compared with the quercetin in high concentration group , the quercetin in low and middle concentration group had more p-Akt （ P〈0.05 ） .Except for the quercetin in high concentration group , theα-SMA expression levels of other four groups were higher than that of the control group（P〈0.05）.The （SiO2 ＋AM） group was higher than the AM group （P〈0.05）.Except for the quercetin in low concentration group , theα-SMA expression levels in middle and high quercetin concentration group were lower than that of the AM group（P〈0.05）.Theα-SMA expression levels of three quercetin concentration groups were lower than that of the （SiO2 ＋AM） group （P〈0.05）.Compared with the quercetin in high concentration group , the α-S

关 键 词：槲皮素 人胚肺成纤维细胞 PI3K/AKT 肌成纤维细胞 二氧化硅 Α-平滑肌肌动蛋白 肺泡巨噬细胞 

分 类 号：R135.2[医药卫生—劳动卫生]