MiR-324-5p可能通过靶向淀粉样前体蛋白抑制棕榈酸诱导的3T3-L1脂肪细胞凋亡  

MiR-324-5p May Inhibit Palmitate-induced Apoptosis of 3T3-L1 Adipocytes by Targeting Amyloid Precursor Proteins

作　　者：王杰 周小敏 杜天宁 刘捷明 史新娥[1] WANG Jie;ZHOU Xiao-Min;DU Tian-Ning;LIU Jie-Ming;SHI Xin-E(Key Laboratory of Animal Genetics,Breeding and Reproduction of Shaanxi Province,Laboratory of Animal Fat Deposition and Muscle Development,College of Animal Science and Technology,Northwest A&F University,Yangling 712100,Shaanxi,China)

机构地区：[1]陕西省动物遗传育种与繁殖重点实验室,西北农林科技大学动物科技学院动物脂肪沉积与肌肉发育实验室,陕西杨凌712100

出　　处：《中国生物化学与分子生物学报》2020年第8期917-923,共7页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家重点基础研究发展(973计划)项目(No.2015CB943102)资助。

摘　　要：miRNAs是一种非编码的小RNA,通过靶向mRNA的3′UTR调控基因的转录后翻译。为明确miR-324-5p对棕榈酸诱导的3T3-L1脂肪细胞凋亡的作用和机制,体外培养3T3-L1脂肪细胞,利用棕榈酸诱导细胞凋亡的同时过表达或抑制miR-324-5p,通过Annexin-V/FITC染色、RT-qPCR等方法检测miR-324-5p对3T3-L1脂肪细胞凋亡的作用。通过在线软件预测miR-324-5p的靶基因并进行验证。结果显示,过表达miR-324-5p能够显著抑制凋亡相关基因BCL-2相关X蛋白(BCL2-associated X protein, Bax)和胱天蛋白酶3(caspase3)的表达水平(P<0.05);而抑制miR-324-5p后能够显著促进这些基因的表达(P<0.05);靶基因预测及验证结果表明,miR-324-5p能够显著降低淀粉样前体蛋白(amyloid precursor protein, APP)的表达水平(P<0.05)。本研究认为,miR-324-5p可能通过靶定APP抑制棕榈酸诱导的3T3-L1脂肪细胞凋亡。miRNAs are non-coding small RNAs that regulate the post-transcriptional translation of genes by targeting the 3′UTR of mRNA. In order to clarify the effect and mechanism of miR-324-5 p on palmitic acid-induced apoptosis of 3 T3-L1 adipocytes, 3 T3-L1 adipocytes were cultured in vitro, and apoptosis was induced by palmitic acid with overexpression or inhibiting miR-324-5 p. The effect of miR-324-5 p on apoptosis of 3 T3-L1 adipocytes was detected by Annexin-V/FITC staining and RT-qPCR. The target gene of miR-324-5 p was searched and verified by online prediction software. Overexpression of miR-324-5 p was found to significantly inhibit the expression of BCL-2-associated X protein(Bax) and Caspase 3. Inhibition of miR-324-5 p significantly promoted the expression of these genes(P<0.05). Target gene prediction and validation results show that miR-324-5 p dramatically reduced the expression level of the amyloid precursor protein(APP)(P <0.05). This study suggests that miR-324-5 p may inhibit palmitate-induced apoptosis of 3 T3-L1 adipocytes by targeting APP.

关 键 词：miR-324-5p 3T3-L1脂肪细胞 凋亡 淀粉样前体蛋白 

分 类 号：Q7[生物学—分子生物学]