关于miR⁃223在慢性乙型肝炎患者外周血中的表达及其生物学作用的研究  

miR ⁃ 223 Expression in the Peripheral Blood of Patients with CHB and ItsBiological Effects

作　　者：徐文超 陈聪[1] XU Wenchao;CHEN Cong(Changzhou Center for Disease Prevention and Control,Changzhou 213022,China)

机构地区：[1]常州市疾病预防控制中心,常州213022

出　　处：《病毒学报》2021年第5期1089-1095,共7页Chinese Journal of Virology

基　　金：江苏省卫生计生委科研课题(项目号:Y2018026),常州市卫生健康青苗人才培养工程资助(项目号:CZQM2020110)。

摘　　要：观察CHB患者与健康人群外周血microRNA(miRNA)差异表达谱,寻找具有抗HBV能力的miRNA。用高通量测序对CHB患者(C组)和健康对照人群(N组)各3例外周血样本进行检测,筛选差异表达的miRNA后进行qRT⁃PCR验证,并在细胞模型中转染miRNA mimic后,ELISA实验观察对HBsAg和HBeAg水平的影响。测序结果共1028个基因,其中上调的基因有27个,下调的基因有20个,无明显差异的基因981个(log2FC<⁃0.585且P<0.05为标准),取FC改变最显著的5个miRNAs进行扩大样本量qRT⁃PCR验证,发现只有miR⁃223表达水平下降83%,与芯片结果保持一致。与未转染相比,在HepG2.2.15细胞中转染miR⁃223 mimic后,miR⁃223表达水平上升5.81倍,而HBsAg和HBeAg分泌分别下降了42%和34%,P<0.05;而转染阴性对照的miR⁃223表达水平及HBsAg和HBeAg分泌无明显改变,P>0.05。miR⁃223在CHB患者外周血中的表达显著下降,基于miR⁃223高表达的新型药物有可能对HBV有较好的抗病毒作用。To ascertain differential microRNA(miR)expression of peripheral blood in chronic hepatitis⁃B(CHB)patients,and to look for miRs that are resistant to the hepatitis⁃B virus(HBV).The whole⁃blood samples of three CHB patients(group C)and a healthy control group(group N)were detected by high⁃throughput sequencing.Differentially expressed miRs were screened for real⁃time reverse transcription⁃quantitative polymerase chain reaction(RT⁃qPCR)verification.After transfection of miR mimics in a cell model,ELISAs were used to observe the effects on levels of the surface antigen of the hepatitis B virus(HbsAg)and hepatitis B viral protein(HbeAg).Sequencing results showed 1028 genes,among which expression of 27 genes was upregulated,20 genes was downregulated,and 981 genes showed no significant difference in expression(log2 FC less than−0.585 and P<0.05 was the standard).Expanded sample⁃size qRT⁃PCR verification of the five miRNAs with the most significant changes in FC revealed that miR⁃223 expression decreased by 83%,a finding that was consistent with the sequencing results.Compared with non⁃transfection,after transfection of a miR⁃223 mimic in HepG2.2.15 cells,miR⁃223 expression increased by 5.81⁃times,whereas secretion of HBsAg and HBeAg decreased by 42%and 34%,respectively(P<0.05).However,miR⁃223 expression and secretion of HBsAg and HBeAg did not change significantly after transfection(P>0.05).miR⁃223 expression in the peripheral blood of CHB patients was significantly lower that that of healthy controls.New drugs based on high expression of miR⁃223 may have a good antiviral effect against HBV infection.

关 键 词：乙型肝炎病毒 慢性乙型肝炎 微小RNA 乙肝表面抗原 乙肝E抗原 

分 类 号：R373.1[医药卫生—病原生物学]