基因沉默促进气道重塑，并诱导过敏性哮喘小鼠的气道平滑肌细胞增殖。
gene silencing contributes to airway remodeling and induces airway smooth muscle cell proliferation in mice with allergic asthma.
作者信息
Wen Xin, Yan Jing, Han Xin-Rui, Zheng Gui-Hong, Tang Ran, Liu Li-Fang, Wu Dong-Mei, Lu Jun, Zheng Yuan-Lin
机构信息
Key Laboratory for Biotechnology on Medicinal Plants of Jiangsu Province, School of Life Science, Jiangsu Normal University, Xuzhou 221116, China.
College of Health Sciences, Jiangsu Normal University, Xuzhou 221116, China.
出版信息
J Thorac Dis. 2018 Jan;10(1):202-211. doi: 10.21037/jtd.2017.12.104.
BACKGROUND
Allergic asthma is a complex genetic disorder that involves interactions between genetic and environmental factors. Usage of PTEN may be a good therapeutic strategy for the management of allergic inflammation. Thus, the present study aims to explore the effects of phosphatase and tensin homolog () gene silencing on airway remodeling and proliferation of airway smooth muscle cells (ASMCs) in a mouse model of allergic asthma.
METHODS
A total of 56 healthy female BABL/c mice (weighing between 16 to 22 grams) were selected and were assigned on random into ovalbumin (OVA; mice were stimulated with OVA to induce allergic asthma), OVA + si-PTEN, normal saline (NS; mice were treated with normal saline) and NS + si-PTEN groups. Masson staining was employed in order to observe lung tissue sections. Immunohistochemical staining was used to detect the expression of α-SMA. Gene silencing was conducted in the NS + si-PTEN and OVA + si-PTEN groups. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blotting were used to detect the mRNA and protein expressions of PTEN in ASMCs of each group. CCK-8 assay and flow cytometry were performed to determine the cell proliferation rate and cell cycle.
RESULTS
Airway remodeling and changes of smooth muscle layer were found in allergic asthmatic mice with thick airway walls. The expression of alpha smooth muscle actin (α-SMA) was significantly higher in ASMCs of the OVA, OVA + si-PTEN and NS + si-PTEN groups compared with ASMCs of the NS group. The mRNA and protein expressions of PTEN reduced in the OVA, OVA + si-PTEN and NS + si-PTEN groups. The rate of ASMCs proliferation in OVA, OVA + si-PTEN and NS + si-PTEN groups were significantly higher than the NS group. The proportion of ASMCs in S and G2 stages increased, while the number of cells in the G1 stage decreased after PTEN gene silencing.
CONCLUSIONS
These results demonstrated that PTEN gene silencing might promote proliferation of ASMCs and airway remodeling in a mouse model of allergic asthma.
背景
过敏性哮喘是一种复杂的遗传疾病，涉及遗传和环境因素之间的相互作用。使用PTEN可能是治疗过敏性炎症的一种良好策略。因此，本研究旨在探讨在过敏性哮喘小鼠模型中，磷酸酶和张力蛋白同源物（PTEN）基因沉默对气道重塑和气道平滑肌细胞（ASMCs）增殖的影响。
方法
共选择56只健康雌性BABL/c小鼠（体重16至22克），随机分为卵清蛋白（OVA；用OVA刺激小鼠诱导过敏性哮喘）组、OVA + si-PTEN组、生理盐水（NS；用生理盐水处理小鼠）组和NS + si-PTEN组。采用Masson染色观察肺组织切片。用免疫组织化学染色检测α-SMA的表达。在NS + si-PTEN组和OVA + si-PTEN组进行基因沉默。采用定量逆转录聚合酶链反应（qRT-PCR）和蛋白质印迹法检测各组ASMCs中PTEN的mRNA和蛋白表达。进行CCK-8测定和流式细胞术以确定细胞增殖率和细胞周期。
结果
在气道壁增厚的过敏性哮喘小鼠中发现气道重塑和平滑肌层变化。与NS组的ASMCs相比，OVA组、OVA + si-PTEN组和NS + si-PTEN组的ASMCs中α平滑肌肌动蛋白（α-SMA）的表达明显更高。OVA组、OVA + si-PTEN组和NS + si-PTEN组中PTEN的mRNA和蛋白表达降低。OVA组、OVA + si-PTEN组和NS + si-PTEN组的ASMCs增殖率明显高于NS组。PTEN基因沉默后，ASMCs在S期和G2期的比例增加，而G1期的细胞数量减少。
结论
这些结果表明，在过敏性哮喘小鼠模型中，PTEN基因沉默可能促进ASMCs增殖和气道重塑。
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