B.A.T. (Investments) Limited, Regents Park Road, Millbrook, Southampton SO15 8TL, UK.
B.A.T. (Investments) Limited, Regents Park Road, Millbrook, Southampton SO15 8TL, UK.
Mutat Res Genet Toxicol Environ Mutagen. 2024 Apr;895:503738. doi: 10.1016/j.mrgentox.2024.503738. Epub 2024 Feb 20.
'Modern' oral tobacco-free nicotine pouches (NPs) are a nicotine containing product similar in appearance and concept to Swedish snus. A three-step approach was taken to analyse the biological effects of NPs and snus extracts in vitro. ToxTracker was used to screen for biomarkers for oxidative stress, cell stress, protein damage and DNA damage. Cytotoxicity, mutagenicity, and genotoxicity were assessed in the following respective assays: Neutral Red Uptake (NRU), Ames and Mouse Lymphoma Assay (MLA). Targeted analysis of phosphorylation signalling and inflammatory markers under non-toxic conditions was used to investigate any potential signalling pathways or inflammatory response. A reference snus (CRP1.1) and four NPs with various flavours and nicotine strengths were assessed. Test article extracts was generated by incubating one pouch in 20 mL of media (specific to each assay) with the inclusion of the pouch material. NP extracts did not induce any cytotoxicity or mutagenic response, genotoxic response was minimal and limited signalling or inflammatory markers were induced. In contrast, CRP1.1 induced a positive response in four toxicological endpoints in the absence of S9: Srxn1 (oxidative stress), Btg2 (cell stress), Ddit3 (protein damage) and Rtkn (DNA damage), and three endpoints in presence of S9: Srxn1, Ddit3 and Rtkn. CRP1.1 was genotoxic when assessed in MLA and activated signalling pathways involved in proliferation and cellular stress and specifically induced phosphorylation of c-JUN, CREB1, p53, p38 MAPK and to a lesser extent AKT1S1, GSK3α/β, ERK1/2 and RSK1 in a dose-dependent manner. CRP 1.1 extracts resulted in the release of several inflammatory mediators including cytokines IL-1α, IL5, IL6, IL8, IL-1RA, MIF and TNF-β, receptor IL-2RA, and growth factors FGF-basic, VEGF and M-CSF. In conclusion these assays contribute to the weight of evidence assessment of the potential comparative health risks of NPs and snus.
“现代”非烟草口含尼古丁袋（NPs）是一种类似外观和概念的含尼古丁产品，类似于瑞典鼻烟。采用三步法分析 NPs 和鼻烟提取物的体外生物学效应。ToxTracker 用于筛选氧化应激、细胞应激、蛋白质损伤和 DNA 损伤的生物标志物。在以下各自的测定中评估细胞毒性、致突变性和遗传毒性：中性红摄取（NRU）、Ames 和小鼠淋巴瘤测定（MLA）。在非毒性条件下，靶向分析磷酸化信号和炎症标志物，以研究任何潜在的信号通路或炎症反应。评估了具有不同口味和尼古丁强度的四种 NPs 与一种参考鼻烟（CRP1.1）。通过将一个烟袋浸泡在 20ml 特定于每种测定的培养基中（包含烟袋材料）来生成测试文章提取物。NPs 提取物不会引起任何细胞毒性或致突变反应，遗传毒性反应最小，并且仅诱导有限的信号或炎症标志物。相比之下，CRP1.1 在无 S9 的情况下在四个毒理学终点中引起了阳性反应：Srxn1（氧化应激）、Btg2（细胞应激）、Ddit3（蛋白质损伤）和 Rtkn（DNA 损伤），以及 S9 存在时的三个终点：Srxn1、Ddit3 和 Rtkn。当在 MLA 中评估时，CRP1.1 具有遗传毒性，并激活了与增殖和细胞应激相关的信号通路，并特异性地诱导 c-JUN、CREB1、p53、p38 MAPK 的磷酸化，以及程度较小的 AKT1S1、GSK3α/β、ERK1/2 和 RSK1，呈剂量依赖性。CRP1.1 提取物导致几种炎症介质的释放，包括细胞因子 IL-1α、IL5、IL6、IL8、IL-1RA、MIF 和 TNF-β、受体 IL-2RA 以及生长因子 FGF-basic、VEGF 和 M-CSF。总之，这些测定有助于评估 NPs 和鼻烟潜在比较健康风险的证据权重。
