miR-202通过抑制PGC1β表达促进3T3-L1前脂肪细胞分化  

miR-202 promotes the differentiation of 3T3-L1 preadipocyte via inhibiting PGC1β expression

作　　者：伍迪 凌宏艳[1] 曹参 何剑琴[1] 杨丝丝[1] 张恺芳[1] 奉水东[3] 

机构地区：[1]南华大学生理学教研室,湖南衡阳421001 [2]南华大学附属第二医院内分泌科,湖南衡阳421001 [3]南华大学社会医学与卫生事业管理学教研室,湖南衡阳421001

出　　处：《基础医学与临床》2018年第1期63-68,共6页Basic and Clinical Medicine

基　　金：教育部留学归国基金[教外司留(2014)1685];湖南教育厅基金(16C1411);衡阳市科技局基金(2016KJ64)

摘　　要：目的观察miR-202对3T3-L1前脂肪细胞分化的影响及可能的机制。方法通过慢病毒感染构建稳定表达AMO-miR-202和乱序对照miRNA细胞系,随后诱导分化。至分化的第9天,油红O染色观察细胞内脂滴的情况;RT-PCR检测过氧化物酶体激活增殖受体γ2(PPARγ2)和a P2的基因表达;Western blot检测PPARγ2、a P2和miR-202靶基因PPARγ辅助活化因子1β(PGC1β)蛋白表达。结果经293T细胞慢病毒包装AMO-miR-202、乱序对照miRNA,荧光显微镜下可见约80%~90%荧光细胞;将上述2组病毒液分别感染3T3-L1前脂肪细胞后,可见约70%~80%荧光细胞。AMO-miR-202组细胞内脂滴及PPARγ2和a P2的mRNA表达显著低于乱序对照组和对照组(P<0.05)。与乱序对照组和对照组相比,AMO-miR-202组PGC1β蛋白表达显著增加(P<0.05),PPARγ2和a P2蛋白表达显著降低(P<0.01),而乱序对照组和脂肪细胞组上述指标无明显差异。结论 miR-202可能通过抑制PGC1β、提高PPARγ2和a P2的表达促进3T3-L1前脂肪细胞分化。Objective To explore the effect and molecular mechanism of miR-202 on the differentiation of 3 T3-L1 preadipocyte.Methods Through lentivirus infected with 3 T3-L1 preadipocytes,we set up the AMO-miR-202 group and the random control group,then,these cells were induced to differentiate,nine days later,differentiation was assessed by Oil Red O staining and we examined the mRNA expression of PPARγ2 and a P2 by RT-PCR method.We examined the mRNA expression of PPARγ2,a P2 and PGC1β by Western blot method. Results After packaging lentivirus with AMO-miR-202 and random sequence control miRNA through cell line 293 T,80%-90% cells with fluorescence were found under fluorescence microscope; After these two lentivirus respectively infected with3 T3-L1 preadipocytes,About 70%-80% cells with fluorescence were found under fluorescence microscope.Oil Red O staining test showed that these cells with Oil Red O stained bright red fat droplets of AMO-miR-202 group andPPARγ2 and a P2 mRNA expression in the AMO-miR-202 group significantly lower than control groups（ P〈0. 05）.Western blot assay showed that the protein expression of PGC1β in the AMO-miRNA-202 group was significantly increased（ P〈0. 05）,but the expression of a P2 and PPARγ2 was significantly decreased（ P〈0. 01）. However,the random control group and the adipocyte group had no significant effect on the above indexes. Conclusions miR-202 can promote the differentiation of 3 T3-L1 preadipocyte by inhibiting the protein expression of PGC1β and improving the protein expression of PPARγ2 and a P2.

关 键 词：miR-202 3T3-L1前脂肪细胞 脂肪细胞分化 PGC1β 

分 类 号：R335.9[医药卫生—人体生理学]