热预适应对大鼠肌肉组织中m^(6)A修饰的影响

Effects of heat precondition on m^(6)A modification in rat muscle tissue

为了解大鼠肌肉中m^(6)A修饰在热预适应过程中的变化,并初步探讨m^(6)A在热预适应过程中发挥的作用.选用24只成年雄性SD大鼠,随机分为2组,12只为预热组,12只为对照组.热预处理组是42℃热处理15 min,每天3次,连续5 d,对照组不做任何处理.处理结束后检测关键酶的转录和翻译表达水平,以及总m^(6)A的变化.与对照组相比,热预处理组的甲基转移酶METTL3和METTL14,KIAA1429的mRNA表达水平均有不同程度上升,METTL3最为显著;去甲基化酶FTO和ALKBH5的mRNA水平下降,FTO蛋白表达水平下降极显著;LC-MS检测发现m^(6)A修饰水平升高.在热预适应过程中,大鼠骨骼肌中m^(6)A甲基化修饰增强,去甲基化减弱,总体m^(6)A水平升高,表明m^(6)A信号通路在大鼠肌肉预适应过程中可能发挥正调控作用.

To observe the changes of m^(6)A modification in rat muscles during heat precondition,and to preliminarily explore the role of m^(6)A in heatprecondition,24 adult male SD rats were randomly divided into 2 groups,12 were preheating group and 12 were control group.The preheating group was treated at 42℃for 15 minutes,3 times a day for 5 consecutive days,and the control group were raised under the room temperature.After the treatment,the transcription and translation expression levels of key enzymes and the changes of total m^(6)A were detected.Compared with the control group,the mRNA expression levels of methyltransferases METTL3 and METTL14 and KIAA1429 in the heat precondition group increased to varying degrees,with METTL3 being the most significant.The mRNA expression of demethylases FTO and ALKBH5 decreased,and the FTO protein reduced significantly.LC-MS detection revealed that the m^(6)A modification level increased in total RNA.In the process of heatprecondition,m^(6)A methylation modification in rat skeletal muscle is enhanced,demethylation is weakened,and overall m^(6)A level increases,indicating that m^(6)A signaling pathway may play a positive regulatory role.