miR-145a-5p调控慢性间歇性低氧诱导脂肪细胞炎症和凋亡的机制  

Mechanism research of miR-145a-5p in the regulation of inflammation and apoptosis in adipocytes induced by chronic intermittent hypoxia

作　　者：徐漫欢[1] 任锡凯 周娈 赵伊敏 余珏珂 苏苗赏[2] XU Manhuan;REN Xikai;ZHOU Luan;ZHAO Yimin;YU Jueke;SU Miaoshang(School of Laboratory Medicine and Life Science,Wenzhou Medical University,Wenzhou 325035,China;Department of Pediatric Respiratory Medicine,the Second Affiliated Hospital&Yuying Children’s Hospital of Wenzhou Medical University,Wenzhou 325027,China)

机构地区：[1]温州医科大学检验医学院(生命科学学院),浙江温州325035 [2]温州医科大学附属第二医院育英儿童医院儿童呼吸科,浙江温州325027

出　　处：《温州医科大学学报》2025年第3期173-178,共6页Journal of Wenzhou Medical University

基　　金：国家自然科学基金项目(81870075);浙江省教育厅一般科研项目(Y202352557);温州市基础性科研项目(Y20210012)。

摘　　要：目的:探究miR-145a-5p基因表达对慢性间歇性低氧(CIH)诱导脂肪细胞炎症和凋亡的影响及分子调控机制,并探索其可能的靶向结合基因通路。方法:体外培养3T3-L1前脂肪细胞,经7 d诱导分化后分为对照组和CIH组,并予相应处理。油红O染色观察细胞形态与脂滴情况;q-PCR法检测miR-145a-5p基因表达;构建miR-145a-5p-mimic转染细胞后,流式细胞仪检测细胞凋亡率;ELISA法检测细胞上清液中IL-1β和IL-10水平;Western blot法检测p-AMPK、成纤维细胞生长因子4(FGF4)及凋亡相关蛋白表达情况。双荧光素酶实验检测miR-145a-5p与FGF4的靶向关系。结果:与对照组相比,CIH组细胞的miR-145a-5p基因表达下降(P<0.05),IL-1β和IL-10水平上升(P<0.05),p-AMPK、FGF4、Bcl-2蛋白表达降低(P<0.05);转染后,转染组与CIH组相比,细胞凋亡率升高(P<0.01),Bcl-2蛋白表达、Bax/Bcl-2比值及FGF4蛋白表达升高(P<0.05),IL-1β水平降低(P<0.05)。双荧光素酶报告FGF4-WT+miR-145a-5p与FGF4-WT相比差异有统计学意义(P<0.01)。结论:miR-145a-5p在脂肪细胞的表达受CIH影响,且对脂肪细胞炎症及凋亡有调节作用,该作用可能通过与FGF4靶向结合而实现。Objective:To explore the effects of miR-145a-5p gene expression on inflammation and apoptosis in adipocytes induced by chronic intermittent hypoxia(CIH),and the possible targeted binding gene pathways.Methods:3T3-L1 preadipocytes were cultured in vitro and differentiated after 7-day induction,which were divided as the Control group and CIH group,and were treated accordingly.Cell morphology and lipid droplets were observed by ORO staining;gene expression of miR-145a-5p was measured by q-PCR;after miR-145a-5p-mimic transfection,the apoptosis rate was measured by flow cytometry;IL-1βand IL-10 levels in the cell supernatant was measured by ELISA;and the expressions of p-AMPK,fibroblast growth factor 4(FGF4)and apoptosis related proteins were measured by Western blot.The targeting relationship of miR-145a-5p and FGF4 was examined in a Dual-luciferase assay.Results:Compared with the Control group,CIH group had decreased gene expression of miR-145a-5p.In CIH group,the levels of IL-1βand IL-10 increased and p-AMPK,FGF4,Bcl-2 protein expression decreased;in Transfection group,the rate of apoptosis,Bcl-2 protein expression,the Bax/Bcl-2 ratio and FGF4 protein expression increased,the expression of IL-1βdecreased compared with CIH group.The dual-luciferase reporter FGF4-WT+miR-145a-5p was significantly different when compared with FGF4-WT.Conclusion:The expression of miR-145a-5p in adipocytes is affected by CIH,and regulates inflammation and adipocyte apoptosis,which may be achieved in combination with FGF4 targeting.

关 键 词：慢性间歇性低氧 脂肪细胞 炎症反应 凋亡 miR-145a-5p 

分 类 号：R725.6[医药卫生—儿科]