Jing Zhenghui, Zhang Haifeng, Wen Yunjie, Cui Shiyu, Ren Yuhua, Liu Rong, Duan Sirui, Zhao Wenbao, Fan Lihong
Department of Pathology of Basic Medicine College, Xi'an Jiaotong University, Xi'an, China.
Institute of Genetics and Developmental Biology of Translational Medicine Institute, Xi'an Jiaotong University, Xi'an, Shaanxi, China.
Front Cell Dev Biol. 2023 May 23;11:1196684. doi: 10.3389/fcell.2023.1196684. eCollection 2023.
Metabolic disorders are an important health concern that threatens life and burdens society severely. ClC-3 is a member of the chloride voltage-gated channel family, and ClC-3 deletion improved the phenotypes of dysglycemic metabolism and the impairment of insulin sensitivity. However, the effects of a healthy diet on transcriptome and epigenetics in ClC-3 mice were not explained in detail. Here, we performed transcriptome sequencing and Reduced Representation Bisulfite Sequencing for the liver of 3 weeks old WT and ClC-3 mice consuming a normal diet to insight into the epigenetic and transcriptomic alterations of ClC-3 deficient mice. In the present study, we found that ClC-3 mice that were younger than 8 weeks old had smaller bodies compared to ClC-3 mice with ad libitum self-feeding normal diet, and ClC-3 mice that were older than 10 weeks old had a similar body weight. Except for the spleen, lung, and kidney, the average weight of the heart, liver, and brain in ClC-3 mice was lower than that in ClC-3 mice. TG, TC, HDL, and LDL in fasting ClC-3 mice were not significantly different from those in ClC-3 mice. Fasting blood glucose in ClC-3 mice was lower than that in ClC-3 mice; the glucose tolerance test indicated the response to blood glucose increasing for ClC-3 mice was torpid, but the efficiency of lowering blood glucose was much higher once started. Transcriptomic sequencing and reduced representation bisulfite sequencing for the liver of unweaned mice indicated that ClC-3 deletion significantly changed transcriptional expression and DNA methylation levels of glucose metabolism-related genes. A total of 92 genes were intersected between DEGs and DMRs-targeted genes, of which Nos3, Pik3r1, Socs1, and Acly were gathered in type II diabetes mellitus, insulin resistance, and metabolic pathways. Moreover, Pik3r1 and Acly expressions were obviously correlated with DNA methylation levels, not Nos3 and Socs1. However, the transcriptional levels of these four genes were not different between ClC-3 and ClC-3 mice at the age of 12 weeks. ClC-3 influenced the methylated modification to regulate glucose metabolism, of which the gene expressions could be driven to change again by a personalized diet-style intervention.
代谢紊乱是一个重要的健康问题，严重威胁生命并给社会带来沉重负担。ClC-3是氯离子电压门控通道家族的成员，ClC-3基因缺失改善了血糖代谢异常的表型以及胰岛素敏感性受损的情况。然而，健康饮食对ClC-3基因敲除小鼠转录组和表观遗传学的影响尚未得到详细阐释。在此，我们对3周龄正常饮食的野生型（WT）和ClC-3基因敲除小鼠的肝脏进行了转录组测序和简化代表性亚硫酸氢盐测序，以深入了解ClC-3基因缺陷小鼠的表观遗传学和转录组学变化。在本研究中，我们发现，与自由采食正常饮食的ClC-3基因敲除小鼠相比，8周龄以下的ClC-3基因敲除小鼠体型较小，而10周龄以上的ClC-3基因敲除小鼠体重相似。除脾脏、肺和肾脏外，ClC-3基因敲除小鼠的心脏、肝脏和大脑平均重量低于ClC-3基因敲除小鼠。禁食的ClC-3基因敲除小鼠的甘油三酯（TG）、总胆固醇（TC）、高密度脂蛋白（HDL）和低密度脂蛋白（LDL）与ClC-3基因敲除小鼠相比无显著差异。ClC-3基因敲除小鼠的空腹血糖低于ClC-3基因敲除小鼠；葡萄糖耐量试验表明，ClC-3基因敲除小鼠对血糖升高的反应迟钝，但一旦开始，降低血糖的效率要高得多。对未断奶小鼠肝脏进行转录组测序和简化代表性亚硫酸氢盐测序表明，ClC-3基因缺失显著改变了葡萄糖代谢相关基因的转录表达和DNA甲基化水平。差异表达基因（DEGs）和差异甲基化区域（DMRs）靶向基因之间共有92个基因相交，其中一氧化氮合酶3（Nos3）、磷脂酰肌醇-3激酶调节亚基1（Pik3r1）、细胞因子信号转导抑制因子1（Socs1）和ATP柠檬酸裂解酶（Acly）聚集在2型糖尿病、胰岛素抵抗和代谢途径中。此外，Pik3r1和Acly的表达与DNA甲基化水平明显相关，而Nos3和Socs1则不然。然而，在12周龄时，这四个基因在ClC-3基因敲除小鼠和ClC-3基因敲除小鼠之间的转录水平没有差异。ClC-3影响甲基化修饰以调节葡萄糖代谢，其中个性化饮食方式干预可再次驱动基因表达发生变化。
