PM2.5诱导大鼠肺组织miRNAs表达谱失调及相关信号通路变化研究  

Imbalance of miRNAs expression profile and changes of related signal pathways in rat lung induced by PM2.5

作　　者：潘柏如 张海军 王杨 杨娟 PAN Boru;ZHANG Haijun;WANG Yang;YANG Juan(Department of Thoracic Surgery, Dazhou Central Hospital, Dazhou 635000, Sichuan, China)

机构地区：[1]达州市中心医院胸外科,四川达州635000

出　　处：《西部医学》2022年第7期985-990,共6页Medical Journal of West China

摘　　要：目的 探讨PM2.5诱导大鼠血清miRNAs表达谱失调及相关信号通路变化。方法 将40只6周龄雄性Wistar大鼠随机分为对照组和PM2.5干预组[低剂量组(2.5 mg/kg)、中剂量组(10 mg/kg)、高剂量组(20 mg/kg)],每组10只。麻醉满意后,PM2.5干预组大鼠分别经气管内滴注PM2.5混悬液2.5、10、20 mg/kg,对照组注射50μL生理盐水,每3 d一次,连续干预70 d。实验结束后处死大鼠,收集肺组织进行病理学检查,并进行miRNA的RNA-seq高通量测序。培养人支气管上皮细胞16 HBE,用100μg/mL浓度的PM2.5处理16 HBE细胞24~48 h,模拟慢性暴露。利用miR-296-5p inhibitor及其阴性对照转染16 HBE细胞12 h后,进行PM2.5干预。Western blot检测Notch信号通路的改变。结果 与对照组比较,PM2.5干预组大鼠肺组织差异表达miRNAs有67个,其中上调48个,下调的19个,表达量差异倍数高于5倍的有4个,即miR-296-5p、miR-27a、miR-182、miR-92a-1-p5,其中miR-296-5p上调了7.85倍,是表达差异最大的miRNA。与对照组比较,经PM2.5干预后,细胞中的Notch1蛋白、HES1蛋白表达量降低,Beclin1蛋白、cleaved-Caspase3蛋白表达量升高(P<0.05);与miRNA阴性对照组相比较,转染miR-296-5p inhibitor后,细胞中的Notch1蛋白,HES1蛋白、Beclin1蛋白的相对表达量均明显升高(P<0.05)。结论 暴露于PM2.5环境中,大鼠肺组织中miRNAs的表达谱发生了紊乱,以miR-296-5p的表达失衡最明显,且miR-296-5p的表达失衡可通过Notch信号通路发挥效应,影响肺上皮细胞功能。Objective To investigate the imbalance of serum miRNAs expression profile and the changes of related signal pathways induced by PM2.5 in rats.Methods 406-week-old male Wistar rats were randomly divided into control group and PM2.5 low/medium/high dose group,once every 3 days for 70 days.Lung tissues were collected for pathological examination,and miRNA RNA SEQ high-throughput sequencing was performed.16 HBE cells were cultured and treated with PM2.5 for 24~48 hours.After 16 HBE cells were transfected with mir-296-5p inhibitor and its negative control for 12 hours,the changes of Notch signaling pathway were detected by Western blotting.Results 67 miRNAs were differentially expressed in the lung tissue of rats in the intervention group,including 48 up-regulated and 19 down regulated,and 4 were more than 5 times higher,namely mir-296-5p,mir-27a,mir-182 and mir-92a-1-p5.Among them,mir-296-5p was up-regulated 7.85 times,which was the miRNA with the largest expression difference.After PM2.5 intervention,the expression of Notch1 protein and Hes1 protein decreased,while the expression of Beclin1 protein and cleaved caspase 3 protein increased(P<0.05);After transfection with mir-296-5p inhibitor,the relative expression of Notch1 protein,Hes1 protein and Beclin1 protein increased significantly(P<0.05).Conclusion Exposed to PM2.5,the expression profile of miRNAs in rat lung tissue is disordered,and the imbalance of mir-296-5p expression is the most obvious,and the imbalance of mir-296-5p expression can affect the function of lung epithelial cells through Notch signal pathway.

关 键 词：肺癌 细颗粒物 MICRORNA NOTCH基因 大鼠 

分 类 号：R734.2[医药卫生—肿瘤]