Manavalan Sridharan, Getachew Bruk, Manaye Kebreten F, Khundmiri Syed J, Csoka Antonei B, McKinley Raechel, Tamas Andrea, Reglodi Dora, Tizabi Yousef
Department of Basic Sciences, National University of Health Sciences, Pinellas Park, FL, USA.
Department of Anatomy, University of Pecs, Pécs, Hungary.
Neurotox Res. 2017 Jul;32(1):8-13. doi: 10.1007/s12640-017-9727-8. Epub 2017 Mar 24.
The detrimental effects of heavy drinking and smoking are multiplied when the two are combined. Treatment modalities for each and especially for the combination are very limited. Although in low concentration, alcohol and nicotine, each may have beneficial effects including neuroprotection, their combination, instead of providing additive protection, may actually lead to toxicity in cell cultures. Pituitary adenylate cyclase-activating polypeptide (PACAP) is an endogenous 38 amino-acid peptide with demonstrated protection against neuronal injury, trauma as well as various endogenous and exogenous toxic agents. The aim of this study was to investigate whether PACAP may also protect against toxicity induced by high alcohol, high nicotine, or the combination of low alcohol and nicotine concentrations, and if so, whether this effect was mediated via PAC1 receptor. We used the neuroblastoma-derived SH-SY5Y cells and applied various colorimetric assays for determination of cell viability or toxicity. Results indicate that PACAP blocks toxicity induced by high alcohol and high nicotine as well as their combination at low concentrations. The effects of PACAP in turn were blocked by the PACAP antagonist (PACAP 6-38), indicating involvement of the PACAP receptor PAC1 and possibly vasoactive intestinal peptide (VIP) receptors in PACAP's protection. Moreover, no combined toxicity of low alcohol and low nicotine could be detected in calcium-free medium. These findings suggest possible beneficial effects of PACAP in preventing alcohol and nicotine toxicity and that calcium contributes to the damage induced by combination of low alcohol and nicotine in SH-SY5Y cells.
大量饮酒和吸烟同时存在时，其有害影响会成倍增加。针对每种行为尤其是两者并存情况的治疗方法非常有限。尽管酒精和尼古丁在低浓度时各自可能具有包括神经保护在内的有益作用，但它们的组合非但不能提供叠加保护，实际上在细胞培养中可能导致毒性。垂体腺苷酸环化酶激活多肽（PACAP）是一种由38个氨基酸组成的内源性肽，已证实其对神经元损伤、创伤以及各种内源性和外源性毒性物质具有保护作用。本研究的目的是调查PACAP是否也能保护细胞免受高浓度酒精、高浓度尼古丁或低浓度酒精与尼古丁组合所诱导的毒性作用，如果可以，这种作用是否通过PAC1受体介导。我们使用源自神经母细胞瘤的SH-SY5Y细胞，并应用各种比色法测定细胞活力或毒性。结果表明，PACAP可阻断高浓度酒精、高浓度尼古丁以及它们低浓度组合所诱导的毒性。PACAP的作用又被PACAP拮抗剂（PACAP 6-38）阻断，这表明PACAP受体PAC1以及可能的血管活性肠肽（VIP）受体参与了PACAP的保护作用。此外，在无钙培养基中未检测到低浓度酒精和低浓度尼古丁的联合毒性。这些发现表明PACAP在预防酒精和尼古丁毒性方面可能具有有益作用，并且钙促成了低浓度酒精和尼古丁组合对SH-SY5Y细胞造成的损伤。
