同济大学学报（医学版）2017，Vol.38Issue(3)：1-7,13,8.DOI:10.16118/j.1008-0392.2017.03.001
MicroRNA-195-5p调节Bmpr1α表达对骨髓间充质干细胞成脂分化的影响
MicroRNA-195-5p affects adipocyte differentiation in bone mesenchymal stem cells by regulating Bmpr1α gene expression
摘要
Abstract
Objective To investigate the effect of rno-miR-195-5p on modulating adipocyte differentiation related to sensitivity for Bushen decoction-medicated serum of bone marrow mesenchymal stem cells (BMSCs) and its mechanism.Methods BMSCs were isolated and cultured in vitro by adherent culture method.The flow cytometric analysis and immunofluorescent staining were used to identify primarily cultured BMSCs in vitro. Quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) was performed for verifying the expression of rno-miR-195-5p in primary BMSCs, intervertebral disc chondrocytes and osteoblasts.Bioinformatics was used for predicting target genes for rno-miR-195-5p.Dual-luciferase reporter assay system and Western blot were performed for verifying the target gene and detecting the expression of Bmpr1α.Oil red O staining was used to assess the differentiation of BMSCs to adipocytes by transfection with miRNA mimics or inhibitors.Results The adherent culture method was practicable to obtain a large number of primary rat BMSCs with typical BMSCs characteristics identified by flow cytometry and immunofluorescent staining.After comparing rno-miR-195-5p expression in Bushen decoction-containing serum culture with that from normal saline control culture,the results of RT-PCR showed that rno-miR-195-5p was markedly up-regulated ( P <0.01) in BMSCs cultured with Bushen decoction-containing serum.Dual luciferase report assay and Western blotting confirmed that rno-miR-195-5p can affect 3′-UTR sequence in Bmpr1α gene.Bioinformatics result showed that Bmpr1α was the direct target gene for rno-miR-195-5p.Over-expression of rno-miR-195-5p in BMSCs significantly suppressed Bmpr1α expression ( P <0.05), and significantly inhibited adipogenic differentiation.We also observed that inhibition of rno-miR-195-5p in BMSCs significantly up-regulated Bmpr1α ( P <0.01), and enhanced adipogenic differentiation.Conclusion The expression of rno-miR-195-5p in rat BMSCs is up-regulated after Bushen decoction-containing serum intervention.rno-miR-195-5p can suppress adipogenic differentiation via regulating Bmpr1α, which may relieve the symptom of osteoporosis.关键词
骨形态发生蛋白受体-1α/rno-miR-195-5p/骨髓间充质干细胞/骨质疏松Key words
bone morphogenetic protein receptor type 1A/rno-miR-195-5p/bone mesenchymal stem cell/osteoporosis分类
医药卫生引用本文复制引用
潘欣,曾思良,梁兴伦,嵇承栋,刘晓东,廖万清..MicroRNA-195-5p调节Bmpr1α表达对骨髓间充质干细胞成脂分化的影响[J].同济大学学报（医学版）,2017,38(3):1-7,13,8.基金项目
国家“九七三”重点基础研究发展计划(2013CB531601) （2013CB531601）
国家自然科学基金(30972633、81173312、81372015) （30972633、81173312、81372015）
上海市卫生和计划生育委员会科研项目(201640253) （201640253）
同济大学附属杨浦医院学科带头人攀登计划(YE2201608) （YE2201608）
