再生障碍性贫血诊断标志物微小RNA-144和微小RNA-451:靶向BHLHE41的机制研究OA

Objective To investigate the expression and clinical significance of microRNA(miR)-144 and miR-451 in the peripheral blood of patients with aplastic anemia(AA).Methods Pe-ripheral blood samples were collected from 35 AA patients in the Department of Hematology of North-ern Jiangsu People's Hospital Affiliated to Yangzhou University from October 2016 to December 2023.The expression levels of miR-144 and miR-451 were detected by quantitative real-time polymerase chain reaction(qRT-PCR).Spearman correlation analysis was used to explore the correlation of the expression levels of miR-144 and miR-451 with laboratory indicators.Logistic regression models were employed to analyze the risk factors for disease severity and the diagnostic value of miR-144 and miR-451 inAA patients.Cox regression analysis was used to investigate the influencing factors of mor-tality in AA patients.Bioinformatics analysis was conducted to screen for differentially expressed genes in AA.The expression level of basic helix-loop-helix family member e41(BHLHE41)in bone marrow mononuclear cells of AA patients was detected by qRT-PCR.Results Among 35 AA pa-tients,there were 17 cases of severe aplastic anemia(SAA)and 18 cases of non-severe aplastic ane-mia(NSAA).The white blood cell(WBC),neutrophil(N),lymphocyte(L),and platelet(PLT)counts in the SAA group were significantly lower than those in the NSAA group(P＜0.05).The rel-ative expression levels of miR-144 and miR-451 in AA patients were 0.49 and 0.52 respectively,which were significantly lower than 1.65 and 1.43 respectively in the control group(healthy ontrols)(P＜0.01).The relative expression levels of miR-144 and miR-451 in both the SAA and NSAA groups were significantly lower than those in the control group(P＜0.01).The expression level of miR-451 was significantly decreased in AA patients with granulocytopenia(N＜0.5 × 109/L,P＜0.05).No significant differences were observed in the expression of miR-144 and miR-451 among AA patients with different genders,ages,degrees of anemia,initial diagnosis status,immunosuppres-sive therapy(IST)regimens,or comorbidities(P＞0.05).The relative expression level of miR-451 was significantly positively correlated with the red blood cell distribution width coefficient of variation(RDW-CV)(r=0.578,P=0.001)and significantly negatively correlated with serum iron(SI)(r=-0.456,P=0.038).Univariate analysis revealed that disease severity and RDW-CV were cor-related with mortality in AA patients(SAA vs NSAA,Log-rankx2=7.658,P=0.006;RDW-CV≥14.8％vs RDW-CV＜14.8％,Log-rank x2=4.048,P=0.044).Further multivariate analysis showed that neither disease severity nor RDW-CV level was a risk factor for mortality in AA patients(disease severity:HR=0.083,95％CI,0.006 to 1.256,P=0.073;RDW-CV:HR=4.653,95％CI,0.651 to 33.241,P=0.125).In the follow-up period,8 AA patients died.No significant differences were observed in the levels of miR-144 and miR-451 between surviving and deceased pa-tients(P＞0.05).Differential gene analysis identified BHLHE41 as one of the upregulated genes.The expression level of BHLHE41 in bone marrow mononuclear cells of AA patients was higher than that in the control group(P＞0.05).Conclusion The levels of miR-144 and miR-451 are de-creased in AA patients,and miR-144 may participate in the pathogenesis of AA by regulating cell dif-ferentiation through targeting BHLHE41.