hsa_circ_0005389在新生儿急性呼吸窘迫综合征细胞炎症模型中的作用研究  

Experimental study on the involvement of hsa_circ_0005389 in the inflammatory process of neonatal acute respiratory distress syndrome

作　　者：游铭钰 周欢 张宇涵 李冰洁[1] 陈筱青[1] YOU Mingyu;ZHOU Huan;ZHANG Yuhan;LI Bingjie;CHEN Xiaoqing(Department of Pediatrics,the First Affiliated Hospital of Nanjing Medical University,Nanjing 210029;Department of Pediatrics,Tangdu Hospital,Air Force Military Medical University,Xi’an 710038,China)

机构地区：[1]南京医科大学第一附属医院儿科,江苏南京210029 [2]空军军医大学附属唐都医院儿科,陕西西安710038

出　　处：《南京医科大学学报（自然科学版）》2023年第7期945-953,共9页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家自然科学基金(81871195)。

摘　　要：目的:基于人环状RNA(circular RNA,circRNA)高通量测序和生物信息学分析结果,研究hsa_circ_0005389与新生儿急性呼吸窘迫综合征(neonatal acuterespiratorydistresssyndrome,NARDS)的关系,以期为NARDS的诊断和治疗提供新的方向。方法:建立脂多糖(lipopolysaccharide,LPS)诱导急性肺损伤细胞模型。通过RT⁃qPCR及Western blot检测炎性标志物及相关通路指标在阴性对照组和敲低hsa_circ_0005389表达的干预组中的表达变化情况,同时用CCK⁃8与流式细胞仪检测正常A549细胞与建立急性肺损伤模型的A549细胞敲低或者过表达hsa_circ_0005389后的增殖与凋亡情况。结果:RT⁃qPCR及Western blot结果显示,A549细胞暴露于10μg/cm^(2)LPS 48 h时,各炎性标志物及相关通路指标高表达(P<0.05)。与阴性对照组相比,敲低hsa_circ_0005389表达后,A549细胞中可溶性肿瘤坏死因子受体1(soluble tumornecrosisfactorreceptor 1,sTNFR1)mRNA相对表达量和肿瘤坏死因子⁃α(tumor necrosisfactorα,TNF⁃α)蛋白表达量显著降低(P<0.05);在急性肺损伤模型中,敲低hsa_circ_0005389表达后,各炎性标志物及相关通路指标mRNA相对表达量均下降(P<0.001)。CCK⁃8与流式细胞仪检测结果显示,与阴性对照组相比,敲低hsa_circ_0005389表达的A549细胞增殖加快,凋亡率降低(P<0.05),而过表达hsa_circ_0005389后A549细胞增殖减慢,凋亡率增加(P<0.05)。结论:急性肺损伤细胞模型中,hsa_circ_0005389促进肺损伤炎性标志物的表达,并且影响肺上皮细胞的增殖和凋亡,提示hsa_circ_0005389参与NARDS的炎症过程,可能是NARDS潜在的干预靶标。Objective:The current study aims to investigate the relationship between hsa_circ_0005389 and neonatal acute respiratory distress syndrome(NARDS)based on high⁃throughput sequencing and bioinformatics analysis of human circular RNA(circRNA)to provide a new direction for the diagnosis and treatment of NARDS.Methods:The acute lung injury model was induced by lipopolysaccharide(LPS).The expression of inflammatory markers and related pathway indexes were detected by RT⁃qPCR and Western blot in the negative control group and the intervention group with knockdown of hsa_circ_0005389 expression.CCK⁃8 and flow cytometry were used to detect the proliferation and apoptosis of blank A549 cells and A549 cells with establishment of acute lung injury model,both of which were knocked⁃down or over⁃expressed hsa_circ_0005389.Result:The mRNA relative expression and the protein expression of inflammatory markers and related pathway indexes were highly expressed and increased significantly after exposed to 10μg/cm^(2)LPS for 48 h(P<0.05).Compared with the negative control group,the relative mRNA expression level of soluble tumor necrosis factor receptor 1(sTNFR1)and the protein expression of tumor necrosis factorα(TNF⁃α)in A549 cells were significantly decreased(P<0.05)after knocking down the expression of hsa_circ_0005389.Inflammatory markers and related pathway indexes decreased significantly(P<0.001)in the acute lung injury model after knocking down the express of hsa_circ_0005389.The proliferation of A549 cells were accelerated and the apoptosis rate was decreased(P<0.05)when the expression of hsa_circ_0005389 was down⁃regulated,while the proliferation of A549 cells was slowed down and the apoptosis rate was increased after hsa_circ_0005389 expression was over⁃regulated(P<0.05)by using CCK⁃8 and flow cytometry as compared with the negative control group.Conclusion:In acute lung injury cell model,hsa_circ_0005389 promoted the expression of inflammatory markers of lung injury,affecting the proliferation and apo

关 键 词：hsa_circ_0005389 新生儿急性呼吸窘迫综合征 炎症 环状RNA 肺损伤 

分 类 号：R563.8[医药卫生—呼吸系统]