Hepatic tissue expression profiles of Acsm3 knockout and control mice.

Acyl-CoA synthetase medium-chain family member 3 (ACSM3) is one member of ACSM family and localized on the outer membrane of mitochondria, which catalyzes the activation of medium-chain (C4-C14) length FAs and xenobiotic carboxylic acids. We found that ACSM3 was lower expressed in the metabolic syndrome (MetS) patients and mice. ACSM3 mainly expressed in the liver. We speculated that ACSM3 deficiency may participated in the MetS progression and then constructed Acsm3 global knockout mice (C57BL/6J background) by CRISPR/Cas9 system. The knockout mice exhibited profound depletion of Acsm3 expression in liver extracts. Male mice are more sensitive to diet-induced MetS than female mice, and thus male mice were used in this study. The phenotype of Acsm3 knockout mice was analyzed with wild type littermates as controls. 12 weeks, livers from three male Acsm3 knockout mice and three male control mice were removed for RNA sequencing. To elucidate the molecular differences in hepatic expression profiles between Acsm3 knockout mice and control mice, at 12 weeks of age, we performed RNA sequencing analysis with three male Acsm3 knockout mice and three male control mice. Before that, all mice were housed under a 12 h light/dark cycle at a temperature of 24°C and relative humidity of (56 ± 10) %, with free access to water and normal diet.中链酰基辅酶A合成酶家族成员3（Acyl-CoA synthetase medium-chain family member 3，ACSM3）属于ACSM家族，定位于线粒体外膜，可催化中链（C4-C14）脂肪酸（fatty acids，FAs）及外源性羧酸的活化反应。本研究发现，代谢综合征（metabolic syndrome，MetS）患者与代谢综合征模型小鼠体内ACSM3的表达水平均显著降低。ACSM3主要在肝脏组织中表达，我们推测ACSM3缺失可能参与代谢综合征的发生发展进程，随后通过CRISPR/Cas9系统构建了C57BL/6J背景的Acsm3全身敲除小鼠。该敲除小鼠的肝脏组织中Acsm3的表达水平被显著敲除。相较于雌性小鼠，雄性小鼠对饮食诱导的代谢综合征更为易感，因此本研究选用雄性小鼠开展实验。本研究以野生型同窝小鼠作为对照，分析Acsm3敲除小鼠的表型。为阐明Acsm3敲除小鼠与野生型对照小鼠的肝脏转录表达谱分子差异，本研究于小鼠12周龄时，分别摘取3只雄性Acsm3敲除小鼠与3只雄性野生型对照小鼠的肝脏组织进行RNA测序分析。在实验开始前，所有小鼠均饲养于12小时光照/黑暗循环、环境温度24℃、相对湿度为(56±10)%的条件下，自由饮水并饲喂正常饲料。