乙酰化修饰在几乎所有的生理过程中都具有广泛的功能作用,如转录和能量代谢。巴豆化修饰主要参与RNA加工,核酸代谢,染色体组装和基因表达,发现巴豆化修饰与乙酰化修饰之间存在竞争关系。先前的研究发现,双氢脂酰脱氢酶(DLD)在白色脂肪组织褐变模型小鼠的棕色脂肪组织(BAT)中高表达,提示DLD与白色脂肪褐变密切相关。本研究通过定量实时PCR(qPCR)进行,蛋白质印迹(WB),酶联免疫吸附测定(ELISA),免疫荧光染色,JC-1染色,Mito-Tracker红色CMXRos染色,油红O染色,Bodipy染色,HE染色,和血脂四重测试。实验表明,DLD促进小鼠白色脂肪组织的褐变。细胞,发现DLD通过RAS/ERK途径激活线粒体功能促进白色脂肪细胞褐变。进一步的研究表明,DLD的巴豆化修饰和乙酰化修饰具有相互抑制作用。同时,DLD巴豆化促进白色脂肪细胞褐变,而DLD乙酰化则相反。最后,蛋白质相互作用分析和免疫共沉淀(Co-IP)测定将Sirtuin3(SIRT3)鉴定为调节DLD的脱乙酰和脱乙酰修饰酶。总之,DLD通过巴豆化修饰和RAS/ERK途径激活线粒体功能促进白色脂肪细胞褐变,为肥胖的控制和治疗提供理论依据,这对未来肥胖及肥胖相关疾病的治疗具有重要意义。
Acetylation modification has a wide range of functional roles in almost all physiological processes, such as transcription and energy metabolism. Crotonylation modification is mainly involved in RNA processing, nucleic acid metabolism, chromosome assembly and gene expression, and it\'s found that there is a competitive relationship between crotonylation modification and acetylation modification. Previous study found that dihydrolipoyl dehydrogenase (DLD) was highly expressed in brown adipose tissue (BAT) of white adipose tissue browning model mice, suggesting that DLD is closely related to white fat browning. This study was performed by quantitative real-time PCR (qPCR), Western blotting (WB), Enzyme-linked immunosorbent assay (ELISA), Immunofluorescence staining, JC-1 staining, Mito-Tracker Red CMXRos staining, Oil red O staining, Bodipy staining, HE staining, and Blood lipid quadruple test. The assay revealed that DLD promotes browning of white adipose tissue in mice. Cellularly, DLD was found to promote white adipocytes browning by activating mitochondrial function through the RAS/ERK pathway. Further studies revealed that the crotonylation modification and acetylation modification of DLD had mutual inhibitory effects. Meanwhile, DLD crotonylation promoted white adipocytes browning, while DLD acetylation did the opposite. Finally, protein interaction analysis and Co-immunoprecipitation (Co-IP) assays identified Sirtuin3 (SIRT3) as a decrotonylation and deacetylation modification enzyme of regulates DLD. In conclusion, DLD promotes browning of white adipocytes by activating mitochondrial function through crotonylation modification and the RAS/ERK pathway, providing a theoretical basis for the control and treatment of obesity, which is of great significance for the treatment of obesity and obesity-related diseases in the future.