Adrenomedullin and Protecting Spinal Motor Neurons Against Doxorubicin-induced Toxicity.

IF 1.1 Q4 NEUROSCIENCES

Basic and Clinical Neuroscience Pub Date : 2024-09-01 DOI:10.32598/bcn.2022.3650.1

Mohammad Ali Takhshid, Amir Mahmoodazdeh, Sayed Mohammad Shafiee, Mohsen Sisakht, Zahra Khosdel

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Abstract

Introduction: In the present study, the culture of embryonic spinal motor neurons (SMNs) was used to assess the impacts of adrenomedullin (AM) on the neurotoxic effects of doxorubicin (DOX).

Methods: To prepare the culture of rat embryonic SMNs, spinal cords were isolated from the rat embryos, digested enzymatically, and triturated to obtain spinal cell suspension. Then, the SMNs were purified from the cell suspension using a single OptiPrep gradient and cultured. The SMNs were treated with DOX (0.0-100 μM) and AM (3.125-100 nM), and their viability and apoptosis were evaluated using MTT and annexin V flow cytometric assays. Oxidative stress was assessed through the measurement of cellular reactive oxygen species (ROS), nitric oxide (NO), malondialdehyde (MDA), and 8-iso-prostaglandin F2α (iPF2α) levels. Finally, qPCR was employed to determine the expressions of interleukin1-β (IL-1β), inducible NO synthase (iNOS), tumor necrosis factor-α (TNF-α), SRY-related protein 9 (SOX9), matrix metalloproteinase (MMP)-3 and -13.

Results: The viability of SMNs decreased following DOX treatment dose-dependently (IC₅₀=10.54 μM). DOX increased the cellular ROS, MDA, NO, and iPF2α levels (P<0.001). Additionally, AM reduced DOX-induced cell death dose-dependently (P<0.001). AM (50 nM) pretreatment also reduced the DOX-induced oxidative stress (P<0.01) and gene expression (P<0.01).

Conclusion: Based on the results, AM might be a protective factor against chemotherapy-induced toxicity in SMNs.

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肾上腺髓质素和保护脊髓运动神经元免受阿霉素诱导的毒性。

在本研究中，采用胚胎脊髓运动神经元（SMNs）培养来评估肾上腺髓质素（AM）对阿霉素（DOX）神经毒性作用的影响。方法：采用从大鼠胚胎中分离脊髓，经酶解、发酵得到脊髓细胞悬浮液的方法制备大鼠胚胎SMNs培养物。然后，使用单一OptiPrep梯度从细胞悬液中纯化SMNs并进行培养。分别用DOX （0.0 ~ 100 μM）和AM （3.125 ~ 100 nM）处理SMNs， MTT和annexin V流式细胞术检测SMNs的细胞活力和凋亡情况。通过测定细胞活性氧（ROS）、一氧化氮（NO）、丙二醛（MDA）和8-异前列腺素F2α (iPF2α)水平来评估氧化应激。最后，采用qPCR检测白细胞介素1-β (IL-1β)、诱导型NO合成酶（iNOS）、肿瘤坏死因子-α (TNF-α)、sry相关蛋白9 （SOX9）、基质金属蛋白酶(MMP)-3、-13的表达。结果：DOX处理后，SMNs的活力呈剂量依赖性下降（IC50=10.54 μM）。DOX增加了细胞ROS、MDA、NO和iPF2α水平(结论：基于这些结果，AM可能是化疗诱导的SMNs毒性的保护因子。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Basic and Clinical Neuroscience NEUROSCIENCES-

CiteScore

2.60

自引率

0.00%

发文量

审稿时长

4 weeks

期刊介绍： BCN is an international multidisciplinary journal that publishes editorials, original full-length research articles, short communications, reviews, methodological papers, commentaries, perspectives and “news and reports” in the broad fields of developmental, molecular, cellular, system, computational, behavioral, cognitive, and clinical neuroscience. No area in the neural related sciences is excluded from consideration, although priority is given to studies that provide applied insights into the functioning of the nervous system. BCN aims to advance our understanding of organization and function of the nervous system in health and disease, thereby improving the diagnosis and treatment of neural-related disorders. Manuscripts submitted to BCN should describe novel results generated by experiments that were guided by clearly defined aims or hypotheses. BCN aims to provide serious ties in interdisciplinary communication, accessibility to a broad readership inside Iran and the region and also in all other international academic sites, effective peer review process, and independence from all possible non-scientific interests. BCN also tries to empower national, regional and international collaborative networks in the field of neuroscience in Iran, Middle East, Central Asia and North Africa and to be the voice of the Iranian and regional neuroscience community in the world of neuroscientists. In this way, the journal encourages submission of editorials, review papers, commentaries, methodological notes and perspectives that address this scope.