To explore the effects of Lyrm1 knockdown on the mitochondrial function of 3 T3-L1 adipocytes using small interfering RNA (siRNA). 3 T3-L1 preadipocytes were infected with either a negative control (NC) expression lentivirus or a Lyrm1-shRNA expression lentivirus and induced to differentiate. The knockdown efficiency of Lrym1-specific shRNA in 3 T3-L1 cells was evaluated by real-time PCR. The ultrastructure of the mitochondria in adipocytes was visualized using transmission electron microscopy after differentiation. The levels of mitochondrial DNA copy numbers and Ucp2 mRNA were detected by real-time quantitative PCR. The levels of ATP production was detected using a photon-counting luminometer. The mitochondrial membrane potential and ROS levels of cells were analyzed with a FACScan flow cytometer using Cell Quest software. Cells transfected with lentiviral-Lyrm1-shRNA showed a significantly reduced transcription of Lyrm1 mRNA compared with NC cells. The size and ultrastructure of mitochondria in Lyrm1 knockdown adipocytes was similar to those of the NC cells. There was no significant difference in mtDNA copy number between the two groups. The total level of ATP production, mitochondrial membrane potential and Ucp2 mRNA expression levels were dramatically increased in adipocytes transfected with Lyrm1 RNAi. Furthermore, the level of ROS was dramatically decreased in Lyrm1 knockdown adipocytes. Knockdown of the Lyrm1 gene in adipocytes resulted in dramatically increased cellular ATP production, mitochondrial membrane potentials and levels Ucp2 mRNA, while ROS levels were significantly decreased. These results imply that mitochondrial function is improved in adipocytes after the knockdown of Lyrm1.

中文翻译：

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Lyrm1 敲低对 3 个 T3-L1 小鼠脂肪细胞线粒体功能的影响。

使用小干扰 RNA (siRNA) 探讨 Lyrm1 敲低对 3 个 T3-L1 脂肪细胞线粒体功能的影响。3 个 T3-L1 前脂肪细胞被阴性对照 (NC) 表达慢病毒或 Lyrm1-shRNA 表达慢病毒感染并诱导分化。通过实时 PCR 评估 Lrym1 特异性 shRNA 在 3 个 T3-L1 细胞中的敲低效率。分化后使用透射电子显微镜观察脂肪细胞中线粒体的超微结构。通过实时定量 PCR 检测线粒体 DNA 拷贝数和 Ucp2 mRNA 的水平。使用光子计数光度计检测 ATP 产生的水平。使用 Cell Quest 软件，用 FACScan 流式细胞仪分析细胞的线粒体膜电位和 ROS 水平。与 NC 细胞相比，用慢病毒-Lyrm1-shRNA 转染的细胞显示出 Lyrm1 mRNA 的转录显着降低。Lyrm1 敲低脂肪细胞中线粒体的大小和超微结构与 NC 细胞相似。两组之间的 mtDNA 拷贝数没有显着差异。在用 Lyrm1 RNAi 转染的脂肪细胞中，ATP 产生的总水平、线粒体膜电位和 Ucp2 mRNA 表达水平显着增加。此外，在 Lyrm1 敲低脂肪细胞中，ROS 水平显着降低。脂肪细胞中 Lyrm1 基因的敲低导致细胞 ATP 产生、线粒体膜电位和 Ucp2 mRNA 水平显着增加，而 ROS 水平显着降低。