时珍国医国药2025，Vol.36Issue(21)：4001-4009,9.DOI:10.70976/j.1008-0805.SZGYGY-2025-2101
益气除痰方调控miR-7-5p/AMBRA1促进弥漫大B细胞淋巴瘤自噬和凋亡的机制研究
A mechanism study on Yiqi Chutan Formula(益气除痰方)in regulating miR-7-5p/AMBRA1 to modulate autophagy and apoptosis in diffuse large B-Cell lymphoma
摘要
Abstract
Objective To investigate the mechanism by which Yiqi Chutan Formula(益气除痰方,YQCT)modulates autophagy and apoptosis in diffuse large B-cell lymphoma(DLBCL).Methods The CCK-8 assay was used to assess the effect of YQCT-containing medicated serum on DLBCL cell viability.Immunofluorescence(IF)was performed to detect autophagy-related protein expression,and flow cytometry(FCM)was used to analyze apoptosis levels.Cell viability and apoptosis-related protein expression were evaluated follow-ing treatment with YQCT-containing medicated serum or the autophagy inhibitor hydroxychloroquine(HCQ).Additionally,a DLBCL tumor-bearing model was established in severe combined immunodeficiency(SCID)mice.These mice were treated with YQCT or HCQ.Levels of autophagy and apoptosis markers,as well as expressions of mTOR,AMBRA1,p-c-Myc,and c-Myc,were assessed to exam-ine the regulatory effect of YQCT on miR-7-5p and potential synergy with HCQ.Results YQCT-containing medicated serum at different concentrations downregulated miR-7-5p,with the most significant effect observed at the 5%concentration(5%MS).Concurrently,AMBRA1 expression was upregulated.The inhibitory effect of 5%MS on cell viability was stronger than that of both the 5%NC control group and the 10%NC control group.IF confirmed that 5%MS significantly induced cell autophagy.CCK-8 and FCM analyses indi-cated that 5%MS inhibited cell growth and increased apoptosis,whereas miR-7-5p mimics reversed the growth inhibitory effect.TUNEL assay revealed that 5%MS increased apoptosis.Furthermore,when combined with HCQ,the pro-apoptotic effect was enhanced in an HCQ dose-dependent manner.Western blot(WB)analysis showed that 5%MS upregulated autophagy-and apoptosis-related pro-tein levels.In contrast,the combination of HCQ with 5%MS resulted in decreased LC3II/I expression,increased p62 expression,and elevated caspase3 levels.IF showed that HCQ downregulated the autophagy-related proteins induced by 5%MS.In vivo,the group treated with YQCT combined with HCQ exhibited the most significant tumor growth inhibition,and HCQ enhanced the pro-apoptotic effect of YQCT.Conclusion YQCT may promote autophagy and apoptosis in DLBCL cells by downregulating miR-7-5p,thereby target-ing AMBRA1.HCQ enhanced the regulatory effect of YQCT on miR-7-5p and shifted the cellular response towards increased apoptosis,contributing to the inhibition of DLBCL cell growth.miR-7-5p may serve as a potential biomarker and therapeutic target for DLBCL.关键词
弥漫大B细胞淋巴瘤/益气除痰方/miR-7-5p/AMBRA1/凋亡/自噬Key words
Diffuse large B-cell lymphoma/Yiqi Chutan Formula(益气除痰方)/miR-7-5p/AMBRA1/Apoptosis/Autophagy分类
医药卫生引用本文复制引用
张翠芬,李志超,杨德宇,陶佳豪,王科,郑创杰,翟林柱..益气除痰方调控miR-7-5p/AMBRA1促进弥漫大B细胞淋巴瘤自噬和凋亡的机制研究[J].时珍国医国药,2025,36(21):4001-4009,9.基金项目
国家自然科学基金(82274604) （82274604）
广州市科学技术计划(202201011748) （202201011748）
北京市希思科临床肿瘤学研究基金会(Y-2020Sciclone/zb/qn-0005) （Y-2020Sciclone/zb/qn-0005）
广州中医药大学"双一流"高水平大学建设学科(中西医结合)后备人才培养项目(A1-2601-22-415-014) （中西医结合）
广州中医药大学第一附属医院2023年中青年骨干人才培养计划 （）
