LYRM1基因沉默对成熟脂肪细胞线粒体形态及其相关基因的影响  

Influences of LYRM1 Knockdown on Mitochondrial Morphology and Mitochondria-Related Genes in Murine Adipocytes

作　　者：朱冠忠[1] 朱金改[1] 史春梅[1] 张敏[1] 曹新国[1] 郭锡熔[1] 

机构地区：[1]南京医科大学附属南京妇幼保健院儿童保健科,南京210004

出　　处：《实用儿科临床杂志》2012年第19期1474-1476,共3页Journal of Applied Clinical Pediatrics

基　　金：国家自然科学基金青年基金(81000348)

摘　　要：目的探讨LYRM1基因沉默对成熟脂肪细胞线粒体形态及相关基因的影响。方法以3T3-L1前体脂肪细胞为工具细胞,建立LYRM1基因沉默细胞株,以转染空载质粒的3T3-L1脂肪细胞作为阴性对照,将前体脂肪细胞诱导分化为成熟脂肪细胞,采用油红O染色观察脂肪细胞分化状态,采用透射电镜观察成熟脂肪细胞的线粒体形态,采用荧光定量PCR技术检测成熟脂肪细胞中线粒体融合基因(Mfn)1/2 mRNA、线粒体动态相关基因(Drp)1 mRNA、线粒体分裂基因(Fis)1 mRNA的表达水平。结果 1.油红O染色诱导分化第8天的脂肪细胞,发现85%以上脂肪细胞已分化为成熟脂肪细胞,细胞形态大而圆,胞质含大量被油红O染成亮红色的脂滴,比较发现2组细胞无论脂滴的大小、数量均无显著差异。2.透射电镜观察发现2组细胞的线粒体形态基本正常,线粒体无明显肿胀、皱缩,线粒体嵴清晰可见,2组细胞间线粒体数目无明显差异。3.LYRM1基因沉默成熟脂肪细胞的Mfn2 mRNA表达水平显著高于对照组成熟脂肪细胞。4.LYRM1基因沉默成熟脂肪细胞的Mfn1 mRNA、Drp1 mRNA和Fis1 mRNA表达水平与对照组成熟脂肪细胞的表达水平无明显差异。结论 LYRM1基因沉默能够引起成熟脂肪细胞Mfn2 mRNA表达水平升高,对线粒体形态结构并无显著影响,提示LYRM1基因沉默可部分影响成熟脂肪细胞线粒体形态相关基因。Objective To explore the effects of LYRM1 gene knockdown on mitochondrial morphology and mitochondria - related genes in routine adipocytes. Methods 3T3 - L1 pre - adipocytes were divided into 2 groups（ negative control group and a LYRMI - RNAi group）. After infected with either a negative control （NC） expression vector or a LYRM1 - shRNA expression vector,pre - adipocytes were induced to differentiate. Cells were stained with oil red O solution,visualized and photographed by light microscopy. The ultrastructure of the mitochondria in murine adipocytes was visualized under transmission electron microscope after differentiation. The expression levels of Mfnl/2 mRNA, Drpl mRNA and Fisl mRNA were detected by real - time quantitative PCR. Results 1. Neither the number of lipid droplets nor their size did not differ between control group and LYRM1 gene knockdown group on the 8th day following stimulation of differentiation. 2. The mitochondria in LYRM1 knockdown adipocytes were similar to those of the negative control cells. The size and ultrastructure of mitochondria were not altered after LYRM1 knockdown. 3. The level of Mfn2 mRNA was dramatically increased in LYRM1 knockdown adipocytes. There was no significant difference in the expression levels of Mfnl mRNA,Drpl mRNA and Fisl mRNA between the 2 groups. Conclusions In murine adipoeytes with LYRM1 gene knockdown,increased expression level of Mfn2 mRNA is not accompanied with changed mitochondrial morphology. It sug- gests that mitochondrial morphology and mitochondria- related genes are partially influenced in adipocytes with LYRM1 gene knockdown in adipocytes.

关 键 词：LYRM1基因 线粒体 3T3-L1脂肪细胞 线粒体融合蛋白 

分 类 号：R723.14[医药卫生—儿科]