微小RNA-26a高表达调控PTEN并促进哮喘大鼠气道平滑肌细胞的增殖  

MicroRNA-26a overespression promotes asthmatic SD rats airway smooth muscle cell proliferation through the regulation of PTEN expression

作　　者：胡彩莉 彭彩霞[1] 胡彩虹[1] Hu Cai-li;Peng Cai-xia;Hu Cai-hong(Department of Geratology,The Second Hospital of Lanzhou University,Lanzhou 730030,China)

机构地区：[1]兰州大学第二医院老年病科,甘肃兰州730030

出　　处：《兰州大学学报（医学版）》2020年第3期43-49,共7页Journal of Lanzhou University（Medical Sciences）

基　　金：兰州大学第二医院萃英计划项目(CY2018-ms11)。

摘　　要：目的探讨微小RNA-26a (miR-26a)对气道平滑肌细胞(ASMC)中PTEN的表达以及细胞增殖的影响和机制。方方法 20只SPF级SD大鼠随机分为正常对照组和哮喘模型组,每组10只。予卵蛋白腹腔注射并雾化吸入制作SD大鼠支气管哮喘模型。苏木精-伊红染色法染色观察2组大鼠肺部组织病理变化。利用荧光定量聚合酶链反应检测2组大鼠气道ASMC中miR-26a的表达。预混10只正常大鼠ASMC细胞,根据不同处理方法分为:空白对照组、转染对照miRNA的阴性对照组、miR-26a转染模拟组、miR-26a抑制剂转染组、PTEN-pGC-Fu-GFP组、pGC-Fu-GFP组和Ly294002组。转染后48 h收集细胞,MTT法检测细胞增殖情况。荧光定量PCR和Western blotting检测空白对照组、阴性对照组、miR-26a转染模拟组和miR-26a抑制剂转染组细胞中PTEN的基因表达和蛋白表达,同时检测PTEN-pGC-FuGFP组、pGC-Fu-GFP组和Ly294002组细胞中细胞周期相关蛋白CyclinD1、p21以及通路相关蛋白p-Akt、JNK1的表达。结果哮喘模型组大鼠ASMC中miR-26a呈现高表达,肺部组织损伤严重。与空白对照组相比,抑制miR-26a表达能够抑制ASMC细胞增殖,上调PTEN表达,过表达miR-26a能够促进ASMC细胞增殖,抑制PTEN表达。与空白对照组比较,过表达PTEN能够抑制ASMC细胞增殖,抑制细胞周期相关蛋白CyclinD1表达,上调细胞周期素依赖性激酶抑制因子p21表达,同时抑制通路蛋白p-Akt和JNK1的表达。结论 miR-26a能够通过影响PTEN的表达,调控ASMC细胞增殖以及PI3K/Akt信号通路。Objective To study the effect of microRNA-26 a on the expression of phosphate and tension homology deleted on chromsome ten(PTEN) and the proliferation of airway smooth muscle cell(ASMC).Methods Twenty SD rats(SPF) were randomly divided into normal control group and asthmatic model group,10 rats per group. The asthmatic model of SD rats were established by immunization with intraperitoneally injected and inhaled ovalbumin. Lung pathological changes were examined using HE staining. The expression of miR-26 a in ASMC cells of two group rats was detected with qRT-PCR. ASMC cells from the ten normal rats were divided into seven groups: control group, negative control group(transfected with no-related miRNA), miR-26 a inhibitor group(transfected with miR-26 a inhibitors), miR-26 a mimic group(transfected with miR-26 a mimic), PTEN-pGC-Fu-GFP group, pGC-Fu-GFP group and Ly294002 group. Cells proliferation was detected by MTT at 48 h after the transfection. The expression of PTEN in the cells of control group,negative control group, miR-26 a inhibitor group and miR-26 a mimic group were detected by real-time PCR and Western blotting methods respectively. The expression of CyclinD1 and p21 as well as p-Akt and JNK1 in the group of control, pGC-Fu-GFP and PTEN-pGC-Fu-GFP were detected by real-time PCR and Western blotting, respectively. Results MiR-26a was overexpressed in ASMC cells of asthmatic model group. Compared with the group of control, down-regulation of miR-26 a obviously inhibited the ASMC cells proliferation and induced the expression of PTEN, while overexpression of miR-26 a promoted the ASMC cells proliferation and inhibited the expression of PTEN. Compared with the group of control, overexpression of PTEN inhibited the ASMC cells proliferation, decreased the expression of CyclinD1, p-Akt and JNK1, and increased the expression of p21. Conclusion MiR-26a could regulate ASMC cells proliferation and the PI3 K/Akt pathways via targeting PTEN expression.

关 键 词：微小RNA-26a 第10号染色体缺失的磷酸酶 气道平滑肌细胞 细胞增殖 调控 PI3K/AKT 

分 类 号：R562.1[医药卫生—呼吸系统]