Mahmoodazdeh Amir, Shafiee Sayed Mohammad, Sisakht Mohsen, Khoshdel Zahra, Takhshid Mohammad Ali
Department of Biochemistry, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.
Autophagy Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.
Iran J Basic Med Sci. 2020 Sep;23(9):1197-1206. doi: 10.22038/ijbms.2020.45134.10514.
Despite effective anticancer effects, the use of doxorubicin (DOX) is hindered due to its cardio and neurotoxicity. The neuroprotective effect of adrenomedullin (AM) was shown in several studies. The present study aimed to evaluate the possible protective effects of AM against DOX-induced toxicity in dorsal root ganglia (DRGs) neurons.
Rat embryonic DRG neurons were isolated and cultured. The effect of various concentrations of DOX (0.0 to 100 µM) in the absence or presence of AM (3.125 -100 nM) on cell death, apoptosis, oxidative stress, expression of tumor necrosis-α (TNF-α), interleukin1- β (IL-1β), inducible nitric oxide synthase (iNOS), matrix metalloproteinase (MMP) 3 and 13, and SRY-related protein 9 (SOX9) were examined.
Based on MTT assay data, DOX decreased the viability of DRG neurons in a dose and time-dependent manner (IC=6.88 µm) while dose-dependently, AM protected DRG neurons against DOX-induced cell death. Furthermore, results of annexin V apoptosis assay revealed the protective effects of AM (25 nm) against DOX (6.88 µM)-induced apoptosis and necrosis of DRG neurons. Also, AM significantly ameliorated DOX-induced oxidative stress in DRG neurons. Real-time PCR results showed a significant increase in the expression of TNF-α, IL-1β, iNOS, MMP 3, and MMP 13, and a decrease in the expression of SOX9 following treatment with DOX. Treatment with AM (25 nM) significantly reversed the effects of DOX on the above-mentioned genes expression.
Our findings suggest that AM can be considered a novel ameliorating drug against DOX-induced neurotoxicity.
尽管阿霉素（DOX）具有有效的抗癌作用，但其心脏和神经毒性阻碍了它的使用。多项研究表明肾上腺髓质素（AM）具有神经保护作用。本研究旨在评估AM对DOX诱导的背根神经节（DRG）神经元毒性的可能保护作用。
分离并培养大鼠胚胎DRG神经元。检测在不存在或存在AM（3.125 - 100 nM）的情况下，不同浓度的DOX（0.0至100 μM）对细胞死亡、凋亡、氧化应激、肿瘤坏死因子-α（TNF-α）、白细胞介素1-β（IL-1β）、诱导型一氧化氮合酶（iNOS）、基质金属蛋白酶（MMP）3和13以及SRY相关蛋白9（SOX9）表达的影响。
基于MTT分析数据，DOX以剂量和时间依赖性方式降低DRG神经元的活力（IC = 6.88 µm），而AM以剂量依赖性方式保护DRG神经元免受DOX诱导的细胞死亡。此外，膜联蛋白V凋亡分析结果显示AM（25 nM）对DOX（6.88 μM）诱导的DRG神经元凋亡和坏死具有保护作用。而且，AM显著改善了DOX诱导的DRG神经元氧化应激。实时PCR结果显示，DOX处理后TNF-α、IL-1β、iNOS、MMP 3和MMP 13的表达显著增加，而SOX9的表达降低。用AM（25 nM）处理显著逆转了DOX对上述基因表达的影响。
我们的研究结果表明，AM可被视为一种对抗DOX诱导的神经毒性的新型改善药物。
