两种不同肺去细胞支架对人肺腺癌A549细胞α平滑肌肌动蛋白表达的影响  

Effect of two different acellular lung matrices on α-SMA expression in A549 cells

作　　者：陈婵[1] 王志翊[2] 翁杰[2] 王志斌[3] 梅劲[3] 杜晓红[4] 王翎[1] 

机构地区：[1]苏州大学附属第一医院博习诊疗中心,215006 [2]温州医科大学附属第二医院育英儿童医院急诊科 [3]温州医科大学基础医学院解剖学教研室 [4]温州医科大学附属第一医院老年医学科

出　　处：《中华医学杂志》2017年第4期303-307,共5页National Medical Journal of China

基　　金：浙江省自然科学基金（LQ17H010004,LYl5H150008）;浙江省卫生厅资助项目（2016KYBl89,2014KYA144）;浙江省温州市公益性科技计划项目（Y20140688）

摘　　要：目的观察正常及纤维化肺去细胞支架对人肺腺癌A549细胞理平滑肌肌动蛋白（α-SMA）表达的影响。方法成年SD大鼠20只，按随机数字表法分为正常对照组与肺纤维化组各10只，肺纤维化组予博来霉素诱导建模，分别制备厚度为500μm的正常去细胞与纤维化肺去细胞支架。根据A549细胞植入的支架不同分成正常支架组和纤维化支架组，单独培养的无支架A549细胞设为无支架对照组，各组分别体外培养1周，免疫荧光染色检测各组A549细胞α-SMA蛋白表达，实时荧光定量-PCR检测细胞α-SMAmRNA表达。结果无支架对照组A549细胞α-SMA免疫荧光染色呈阳性表达，正常支架组与肺纤维化支架组免疫荧光染色呈弱阳性表达，其中肺纤维化支架组α-SMA免疫荧光强度显著低于正常支架组（P〈0．05）；正常支架组、肺纤维化支架组α-SMA mRNA相对表达量均显著低于无支架对照组（0．70±0．11、0．55±0．12比1．28±0．21），且肺纤维化支架组显著低于正常支架组（均P〈0．05）。结论正常及纤维化肺去细胞支架在体外可不同程度下调A549细胞α-SMA表达，其中纤维化肺去细胞支架作用尤为明显。Objective To explore the effect of acellular normal and fibrotic lung matrices on alpha smooth muscle actin （ct-SMA） expression in human lung adenoearcinoma cell line A549. Methods Twenty adult SD rats were randomly divided into normal group and idiopathic pulmonary fibrosis （IPF）group （n = 10 each）. The pulmonary fibrosis was induced by Bleomycin. Normal and fibrotic decellularized lungs were made, then sections with 500 μm thick were cut by a standard Vibratomc. None scaffold was set as control group. A549 cells were seeded dropwise into different slices （normal and fibrotic scaffolds ）, and cultured for one week in vitro. The expression of α-SMA was measured by immunofluorescence staining and quantitative real time polymerase chain reaction （qRT-PCR）. Results In control group, the expression of ct-SMA protein was positive in A549 cells by immnnofluorescence staining. However, it expressed weakly both in normal and fibrotic scaffold group, and the fluorescence intensity in fibrotic scaffold group was significant lower than that in normal group （ P 〈 0. 05 ）. The relative expression amount of α-SMA mRNA in normal and fibrotie scaffold group were （0. 70 ± 0. 11 ） and （0. 55 ±0. 12） , which were significant lower than that of control group （ 1.28±0. 21 ） （P 〈0. 05）. Moreover, the relative expression of α-SMA mRNA in fibrotic scaffold group was decreased compared to that in normal scaffold group （P 〈 0.05 ）. Conclusions Acellular normal and fibrotic lung scaffold can downregulate the expression of α-SMA in human lung adenocarcinoma cell line A549. It may inhibit the movement of A549 cells in acellular normal and fibrotic lung matrices, especially in acellular fibrotic lung scaffold.

关 键 词：肺纤维化 细胞骨架 去细胞 肺肿瘤 Α-平滑肌肌动蛋白 

分 类 号：R734.2[医药卫生—肿瘤]