Effect of SOD1K123R mutation on gene expression in nucleus pulposus cells treated with lactate.

Among musculoskeletal disorders, intervertebral disc degeneration (IVDD) is the primary cause of discogenic pain and associated disability and productivity loss in the workforce worldwide.Oxidative stress is an important inducing and aggravating factor of intervertebral disc degeneration. We found that lactate could induce the lactylation of SOD1 lysine 123 in nucleus pulposus cells, which impaired the antioxidant ability of the enzyme. Therefore, we mutated lysine (Lys) to arginine (Arg) at residue 123 of SOD1 in nucleus pulposus cells to eliminate the lactylation at residue 132 and investigated the underlying mechanism by RNA sequencing. Overall design: After knocking down SOD1 in nucleus pulposus cells, RNA-seq was performed after transfection of wild-type SOD1 and K123R mutant SOD1 with 10 mM lactate for 7 days

在各类肌肉骨骼疾病中，椎间盘退变（intervertebral disc degeneration, IVDD）是全球范围内引发椎间盘源性疼痛，继而导致劳动者残疾与生产力损失的首要病因。氧化应激是椎间盘退变的重要诱发与加重因素。本研究发现，乳酸可诱导髓核细胞中超氧化物歧化酶1（superoxide dismutase 1, SOD1）第123位赖氨酸残基发生乳酸化修饰，进而削弱该酶的抗氧化能力。为此，我们在髓核细胞中对SOD1第123位赖氨酸残基进行精氨酸（Arg）突变，以消除第132位残基的乳酸化修饰，并通过RNA测序（RNA sequencing）探究其潜在分子机制。实验整体设计：在髓核细胞中敲低SOD1后，分别转染野生型SOD1与K123R突变型SOD1，经10 mM乳酸处理7天后，开展RNA测序。