甘丙肽对二甲萘醌引起的人肾胚A型细胞株氧化损伤的保护作用及机制

Protective effect of galanin on DMNQ-induced oxidative stress and cell damage in HEK-293A cells

目的探讨甘丙肽（GAL）对二甲萘醌（DMNQ）引起的人肾胚A型细胞株（HEK-293A）细胞氧化损伤的保护作用及生物学机制。方法将HEK-293A培养细胞分为对照组，DMNQ药物组以及DMNQ＋GAL／AR—M1896药物组，利用常规聚合酶链反应（RT—PCR）方法检测HEK-293A细胞内GAL及其不同受体的mRNA表达水平，MTS方法检测不同药物处理后HEK-293A的细胞活性。结果HEK-293A细胞GAL及2型受体（GalR2）的mRNA表达水平较高。同时，用不同浓度DMNQ处理下发现存在剂量相关毒性效应，IC50值约为13．4uM。利用15uM DMNQ处理细胞，同时孵育1uM及100nMGAL可以显著改善DMNQ引起的细胞活力降低，细胞活力较DMNQ处理组分别提高24．4％、18．8％。GalR2激动剂AR—M18961uM及100uM也可得到与GAL相似的作用，细胞活力较DMNQ处理组分别提高8．7％、8．9％。结论GAL对DMNQ引起的HEK-293A细胞氧化损伤具有保护作用，这种作用可能通过GalR2发挥效应。

Objective To study the protective effect of galanin on 2,3-Dimethoxy-1 ,4-naphthoquinone （DMNQ）-induced oxidative stress and cell damage in HEK-293A eells and its possible mechanisms. Methods The expression of galanin and its three receptors （GalR1-3） in HEK-293A were determined with RT- PCR technique. The cultured HEK-293A cells were divided into four groups：Control, DMNQ, DMNQ ＋ GAL and DMNQ ＋ AR-M1896 and the cell viability was measured with CellTiter 96 AQueous One Solution Cell Proliferation Assay （MTS）. Results The RT-PCR data revealed the presence of gaianin and its receptors in HEK-293A cells. The expression level was in the order of GalR2 = Galanin 〉 GalR3 〉 GalR1. DMNQ caused oxidative stress and cell damage in HEK-293A cells with a dose-dependent manner with an IC50 = 13.4 uM. Application of galanin reduced the DMNQ-induced cellular toxicity in HEK-293A, which increased cell viability by 24.4% and 18.8% in 1 uM and 100 nM, respectively. AR-M1896,an agonist of GalR2 had a similar effect,increased cell viability by 8.7% and 8.9% in 1 uM and 100 nM, respectively. Conclusion These data suggest that galanin has a proteetive effect on DMNQ-induced oxidative stress and cell damage in HEK-293A cells, probably mediated by CalR2.