circFADS2靶向miR-125a-5p调控MPP+诱导的SK-N-SH细胞损伤的机制  

Mechanism of circFADS2 targeting miR-125a-5p to regulate SK-N-SH cell injury induced by MPP+

作　　者：张永志[1] 顾平(指导)[1] 王文婷[1] 刘惠苗[1] 张赛[1] 刘义晗 ZHANG Yongzhi;GU Ping;WANG Wenting;LIU Huimiao;ZHANG Sai;LIU Yihan(Second Department of Neurology,the First Hospital of Hebei Medical University,Shijiazhuang 050031,China)

机构地区：[1]河北医科大学第一医院神经内二科,石家庄050031

出　　处：《中国免疫学杂志》2023年第6期1243-1247,共5页Chinese Journal of Immunology

基　　金：河北省卫生健康委员会资助项目(20210310);河北省科技厅民生科技专项河北省重点研发计划资助项目(20377721D)。

摘　　要：目的:探究circFADS2靶向miR-125a-5p调控帕金森病(PD)细胞炎症反应和凋亡的机制。方法:100μmol/L MPP+诱导SK-N-SH细胞建立PD模型,分为Con组、PD组、PD+pcDNA组、PD+pcDNA-circFADS2组、PD+anti-miR-NC组、PD+anti-miR-125a-5p组、PD+pcDNA-circFADS2+miR-NC组、PD+pcDNA-circFADS2+miR-125a-5p组。qRT-PCR检测circFADS2和miR-125a-5p表达;ELISA检测IL-1β、TNF-α含量;流式细胞术检测细胞凋亡;Western blot检测cleaved-caspase3、cleaved-caspase9蛋白表达;双荧光素酶报告实验检测circFADS2与miR-125a-5p的靶向关系。结果:与Con组相比,PD组circFADS2表达降低,miR-125a-5p表达、凋亡率、IL-1β、TNF-α含量、cleaved-caspase3、cleaved-caspase9蛋白表达升高(P<0.05)。circFADS2过表达或干扰miR-125a-5p表达可降低MPP+诱导的SK-N-SH细胞凋亡率、IL-1β、TNF-α含量、cleaved-caspase3、cleaved-caspase9蛋白表达(P<0.05)。circFADS2靶向负调控miR-125a-5p表达,上调miR-125a-5p表达逆转了circFADS2过表达对PD细胞炎症反应和凋亡的作用。结论:circFADS2过表达通过降低miR-125a-5p表达抑制PD细胞炎症反应和凋亡。Objective:To explore mechanism of circFADS2 targeting miR-125a-5p to regulate inflammation and apoptosis of Parkinson's disease(PD)cells.Methods:100μmol/L MPP+was used to induce SK-N-SH cells to establish a PD model,and divided into Con group,PD group,PD+pcDNA group,PD+pcDNA-circFADS2 group,PD+anti-miR-NC group,PD+anti-miR-125a-5p group,PD+pcDNA-circFADS2+miR-NC group and PD+pcDNA-circFADS2+miR-125a-5p group.qRT-PCR was used to detect expressions of circFADS2 and miR-125a-5p;ELISA was used to detect levels of IL-1βand TNF-α;flow cytometry was uesd to detect cell apoptosis;Western blot was used to detect protein expressions of cleaved-caspase3 and cleaved-caspase9;dual luciferase reporter experiment was used to detect targeting relationship between circFADS2 and miR-125a-5p.Results:Compared with Con group,expression of circ‑FADS2 in PD group was decreased,and expression of miR-125a-5p,apoptosis rate,IL-1β,TNF-αcontents,cleaved-caspase3 and cleaved-caspase9 protein expressions were increased(P<0.05).Overexpression of circFADS2 or interference with miR-125a-5p reduced MPP+-induced SK-N-SH cell apoptosis rate,IL-1β,TNF-αcontents,cleaved-caspase3 and cleaved-caspase9 protein expressions(P<0.05).circFADS2 targeting and negatively regulated miR-125a-5p expression,and up-regulating expression of miR-125a-5p reversed effect of circFADS2 overexpression on inflammation and apoptosis of PD cells.Conclusion:Overexpression of circFADS2 inhibits inflammatory response and apoptosis of PD cells by reducing expression of miR-125a-5p.

关 键 词：circFADS2 miR-125a-5p 帕金森病 炎症反应 凋亡 

分 类 号：R742.5[医药卫生—神经病学与精神病学]