Lv Zheng, Yin Shao, Cheng Ziguan, Wang Kekai
Department of Orthopedics, Hospital of Chengdu University of Traditional Chinese Medicine, Chengdu, Sichuan 610032, P.R. China.
Anorectal Department, Hospital of Chengdu University of Traditional Chinese Medicine, Chengdu, Sichuan 610032, P.R. China.
Exp Ther Med. 2022 Jun;23(6):421. doi: 10.3892/etm.2022.11348. Epub 2022 Apr 29.
Lenalidomide (LEN) has been reported to exert antitumor, anti-inflammatory, anti-angiogenic and immunomodulatory activities. However, the effects of LEN in spinal cord injury (SCI) are yet to be fully elucidated. The present study was conducted to identify whether LEN has a healing effect on SCI and to determine the underlying mechanism of action. The viability of HO-stimulated PC12 cells was detected using a Cell Counting Kit-8 assay. The viability of PC12 cells treated with LEN was examined with an MTT assay. The level of lactate dehydrogenase (LDH) was measured using an LDH assay kit, while the levels of the oxidative stress-related factors malondialdehyde, superoxide dismutase, glutathione peroxidase and catalase in PC12 cells were determined using commercial kits. Oxidative stress-related proteins were examined via western blotting. PC12 cell apoptosis was observed using a TUNEL assay, while apoptosis markers and Notch signaling-related proteins were examined via western blotting. The results of the present study demonstrated that HO decreased PC12 cell viability in a dose-dependent manner. However, treatment with LEN significantly improved the viability of HO-stimulated PC12 cells and alleviated HO-induced cytotoxic injury. Additionally, LEN treatment inhibited the oxidative stress and apoptosis induced by HO in PC12 cells. Notch signaling-related protein expression was downregulated after LEN treatment in HO-stimulated PC12 cells. In addition, a Notch agonist reversed the effects of LEN on HO-induced oxidative stress and PC12 cell apoptosis. In summary, it was demonstrated that LEN alleviated the HO-induced injury of PC12 cells by blocking the Notch signaling pathway, suggesting the value of applying LEN to the treatment of SCI.
据报道，来那度胺（LEN）具有抗肿瘤、抗炎、抗血管生成和免疫调节活性。然而，LEN在脊髓损伤（SCI）中的作用尚未完全阐明。本研究旨在确定LEN是否对SCI具有修复作用，并确定其潜在的作用机制。使用细胞计数试剂盒-8检测HO刺激的PC12细胞的活力。用MTT法检测LEN处理的PC12细胞的活力。使用乳酸脱氢酶（LDH）检测试剂盒测量LDH水平，而使用商业试剂盒测定PC12细胞中氧化应激相关因子丙二醛、超氧化物歧化酶、谷胱甘肽过氧化物酶和过氧化氢酶的水平。通过蛋白质印迹法检测氧化应激相关蛋白。使用TUNEL法观察PC12细胞凋亡，而通过蛋白质印迹法检测凋亡标志物和Notch信号相关蛋白。本研究结果表明，HO以剂量依赖性方式降低PC12细胞活力。然而，LEN处理显著提高了HO刺激的PC12细胞的活力，并减轻了HO诱导的细胞毒性损伤。此外，LEN处理抑制了HO诱导的PC12细胞氧化应激和凋亡。在HO刺激的PC12细胞中，LEN处理后Notch信号相关蛋白表达下调。此外，Notch激动剂逆转了LEN对HO诱导的氧化应激和PC-12细胞凋亡的影响。总之，证明LEN通过阻断Notch信号通路减轻了HO诱导的PC12细胞损伤，提示应用LEN治疗SCI的价值。
