海南医科大学学报2025，Vol.31Issue(11)：840-848,9.DOI:10.13210/j.cnki.jhmu.20250314.005
miR-126a-3p靶向Bcl-6介导凋亡途径调控脓毒症Th1/Th2细胞平衡
miR-126a-3p targets Bcl-6 mediates apoptosis pathway to regulate Th1/Th2 cell balance in sepsis
摘要
Abstract
Objective:To explore the relationship between miR-126a-3p and B-cell lymphoma 6(Bcl-6)as well as its effects and mechanisms on T-lymphocyte apoptosis and T-cell differentiation in mice with sepsis.Methods:The miRWalk database was used to predict the target genes of miR-126a-3p,and the Cytoscape software was further used to screen the core target genes.Mice were then randomly divided into normal control group(the sham group),sepsis group(the LPS group),LPS+miR-126a-3p mimic group(the miR-126 mimic group),and LPS+miR-126a-3p inhibitor group(the miR-126 inhibitor group)by intraperitone-al injection of 0.9%sodium chloride solution(NS)and lipopolysaccharide(LPS;20 mg/kg).Peripheral blood was collected from the tail vein of mice after 48 hours.T lymphocytes were extracted by density gradient centrifugation,and then miR-126a-3p mimic and inhibitor were transfected into septic T lymphocytes of miR-126 mimic group and miR-126 inhibitor group.Reverse transcription-quantitative polymerase chain reaction(RT-qPCR)and Western blotting were used to detect Bcl-6 mRNA and pro-tein levels in T cells,Annexin V-FITC/PI double staining method was used to detect apoptosis of T lymphocytes,flow cytome-try was used to detect the helper T cell(Th)1,Th2 ratio,and finally the data were statistically analysed by GraphPad Prism 9.0 software.Results:A total of 5 930 predicted targets of miR-126a-3p were retrieved from the miRWalk database,of which Bcl-6 was one of the core targets of miR-126a-3p.The expression level of Bcl-6 was positively correlated with the expression level of miR-126a-3p,and the two had a targeted regulatory relationship.And compared with the sham group,the LPS group showed a de-crease in Bcl-6 mRNA and protein expression,an increase in T-lymphocyte apoptosis,an increase in T-cell differentiation towards Th1,a decrease in differentiation towards Th2,and an increase in the Th1/Th2 ratio,with the difference being statistically signifi-cant in all cases(P<0.001);Compared with the LPS group,after enhancing(miR-126 mimic group)or inhibiting(miR-126 in-hibitor group)miR-126a-3p expression,Bcl-6 mRNA and protein expression,T-cell apoptosis,the ratio of Th1 and Th2 cells showed corresponding increases or decreases,and the differences were statistically significant(P<0.01).Conclusion:The expres-sion level of the anti-apoptotic protein Bcl-6 in T lymphocytes of sepsis mice is abnormal,and its expression level is regulated by miR-126a-3p,which affects the apoptosis of T lymphocytes and the differentiation of Th1/Th2 cells,thereby regulating the im-mune status of sepsis.This study can provide new therapeutic targets and theoretical basis for the treatment of patients with sepsis.关键词
脓毒症/miR-126a-3p/B细胞淋巴瘤6/细胞凋亡/T淋巴细胞Key words
Sepsis/miR-126a-3p/B-cell lymphoma 6/Apoptosis/T-lymphocyte分类
临床医学引用本文复制引用
李苗苗,王洁,齐雅囡,王婷,王德伟,刘成..miR-126a-3p靶向Bcl-6介导凋亡途径调控脓毒症Th1/Th2细胞平衡[J].海南医科大学学报,2025,31(11):840-848,9.基金项目
This study was supported by Natural Science Program of Anhui Provincial Colleges and Universities(2023AH051996) （2023AH051996）
Natural Science Program of Bengbu Medical College(2022byzd039) 安徽省高等学校自然科学基金资助项目(2023AH051996) （2022byzd039）
蚌埠医学院自然科学基金资助项目(2022byzd039) （2022byzd039）
