Department of Pathology, Hebei Medical University, Shijiazhuang, China.
Center of Metabolic Diseases and Cancer Research, Institute of Medical and Health Science of Hebei Medical University, Shijiazhuang, China.
Cell Death Dis. 2021 Jun 23;12(7):642. doi: 10.1038/s41419-021-03930-2.
Diabetic peripheral neuropathy (DPN) is the most common complication of diabetes mellitus (DM) and the dysfunction of Schwann cells plays an important role in the pathogenesis of DPN. Thioredoxin-interacting protein (TXNIP) is known as an inhibitor of thioredoxin and associated with oxidative stress and inflammation. However, whether TXNIP is involved in dysfunction of Schwann cells of DPN and the exact mechanism is still not known. In this study, we first reported that TXNIP expression was significantly increased in the sciatic nerves of diabetic mice, accompanied by abnormal electrophysiological indexes and myelin sheath structure. Similarly, in vitro cultured Schwann cells TXNIP was evidently enhanced by high glucose stimulation. Again, the function experiment found that knockdown of TXNIP in high glucose-treated RSC96 cells led to a 4.12 times increase of LC3-II/LC3-I ratio and a 25.94% decrease of cleaved caspase 3/total caspase 3 ratio. Then, DNA methyltransferase (DNMT) inhibitor 5-Aza has been reported to benefit Schwann cell in DPN, and here 5-Aza treatment reduced TXNIP protein expression, improved autophagy and inhibited apoptosis in high glucose-treated RSC96 cells and the sciatic nerves of diabetic mice. Furthermore, DNMT1 and DNMT3a upregulation were found to be involved in TXNIP overexpression in high glucose-stimulated RSC96 cells. Silencing of DNMT1 and DNMT3a effectively reversed high glucose-enhanced TXNIP. Moreover, high glucose-inhibited PI3K/Akt pathway led to DNMT1, DNMT3a, and TXNIP upregulation in RSC96 cells. Knockdown of DNMT1 and DNMT3a prevented PI3K/Akt pathway inhibition-caused TXNIP upregulation in RSC96 cells. Finally, in vivo knockout of TXNIP improved nerve conduction function, increased autophagosome and LC3 expression, and decreased cleaved Caspase 3 and Bax expression in diabetic mice. Taken together, PI3K/Akt pathway inhibition mediated high glucose-induced DNMT1 and DNMT3a overexpression, leading to cell autophagy inhibition and apoptosis via TXNIP protein upregulation in Schwann cells of DPN.
糖尿病周围神经病变(DPN)是糖尿病(DM)最常见的并发症,雪旺细胞功能障碍在 DPN 的发病机制中起着重要作用。硫氧还蛋白相互作用蛋白(TXNIP)是一种硫氧还蛋白抑制剂,与氧化应激和炎症有关。然而,TXNIP 是否参与 DPN 雪旺细胞的功能障碍以及确切机制尚不清楚。在这项研究中,我们首先报道了 TXNIP 表达在糖尿病小鼠的坐骨神经中显著增加,同时伴有异常的电生理指标和髓鞘结构。同样,体外培养的雪旺细胞在高葡萄糖刺激下 TXNIP 明显增强。此外,功能实验发现,高葡萄糖处理的 RSC96 细胞中 TXNIP 的敲低导致 LC3-II/LC3-I 比值增加 4.12 倍,裂解的 caspase 3/总 caspase 3 比值降低 25.94%。然后,已报道 DNA 甲基转移酶(DNMT)抑制剂 5-Aza 有益于 DPN 中的雪旺细胞,在这里,5-Aza 处理降低了 TXNIP 蛋白表达,改善了高葡萄糖处理的 RSC96 细胞和糖尿病小鼠坐骨神经中的自噬并抑制了细胞凋亡。此外,发现 DNMT1 和 DNMT3a 的上调参与了高葡萄糖刺激的 RSC96 细胞中 TXNIP 的过表达。DNMT1 和 DNMT3a 的沉默有效地逆转了高葡萄糖增强的 TXNIP。此外,高葡萄糖抑制的 PI3K/Akt 通路导致 RSC96 细胞中 DNMT1、DNMT3a 和 TXNIP 的上调。DNMT1 和 DNMT3a 的敲低阻止了 RSC96 细胞中 PI3K/Akt 通路抑制引起的 TXNIP 上调。最后,体内敲除 TXNIP 改善了糖尿病小鼠的神经传导功能,增加了自噬小体和 LC3 的表达,并降低了裂解的 Caspase 3 和 Bax 的表达。总之,PI3K/Akt 通路抑制介导高葡萄糖诱导的 DNMT1 和 DNMT3a 过表达,导致 DPN 雪旺细胞中自噬抑制和细胞凋亡通过 TXNIP 蛋白上调。