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目的初步探讨miR-7对LXRβ的抑制作用及其在肝细胞脂质生成中的作用。方法生物信息学分析,预测LXRβmRNA的3’-UTR上miR-7的可能结合位点。在人肝细胞系Hep G2中转染miR-7,以Real-time PCR、Western blot检测LXRβmRNA水平和蛋白水平表达的变化;构建LXRβ3’-UTR报告基因质粒p MIR/LXRβ,Report assay检测报告基因荧光素酶活性;Real-time PCR检测miR-7对LXRβ激动剂诱导的脂质生成相关靶基因FAS、ACC的表达变化。结果在人肝细胞中,miR-7能够显著抑制LXRβ的表达(P<0.05),且miR-7能降低LXRβ重组质粒p MIR/LXRβ的荧光素酶活性(P<0.05)。同时抑制其激动剂对脂质代谢相关靶基因的诱导作用。结论 miR-7可通过结合在LXRβ3’-UTR区域而抑制其表达,进而抑制FAS、ACC的表达而抑制脂质生成。
Objective To investigate the inhibitory effect of miR-7 on LXRβ and its role in hepatocyte lipid production. Methods Bioinformatic analysis predicted the possible binding sites for miR-7 on the 3’-UTR of LXRβ mRNA. MiR-7 was transfected into Hep G2 cell line, and the changes of LXRβmRNA and protein expression were detected by Real-time PCR and Western blot. The LXRβ3’-UTR reporter plasmid p MIR / LXRβ was constructed and the reporter assay Luciferase activity. Real-time PCR was used to detect the expression of FAS and ACC in LXRβ-induced lipid-related target genes. Results In human hepatocytes, miR-7 significantly inhibited the expression of LXRβ (P <0.05), and miR-7 decreased the luciferase activity of LXRβ recombinant plasmid p MIR / LXRβ (P <0.05). While inhibiting its agonist on lipid metabolism related target gene induction. Conclusions miR-7 can inhibit the expression of FAS and ACC by inhibiting the expression of miR-7 in the 3’-UTR region of LXRβ and inhibiting the lipid production.