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目的采用体外细胞毒性检测方法研究亚硝酸钠(NaNO2)对L929细胞增殖的影响,探讨在此过程中遗传物质的损伤情况。方法体外培养L929细胞,用溴化四氮唑蓝法检测不同浓度NaNO2对细胞增殖的影响;并采用单细胞凝胶电泳技术及KCl-十二烷基磺酸钠沉淀法(K-SDS)检测NaNO2对L929细胞DNA损伤及DNA-蛋白质交联的作用。结果 NaNO2处理L929细胞48h后,细胞增殖出现毒物兴奋效应(hormesis),其剂量范围约为0～0.0384mg/L;染毒剂量为0.02、1 500 mg/L时,细胞尾长分别为(2.72±1.99)、(3.26±3.09)μm,尾DNA%分别为(7.87±6.63)%及(7.75±4.90)%,Olive尾矩分别为(0.80±0.64)及(0.79±0.64),与阴性对照组〔(1.83±1.08)μm、(6.07±3.59)%、(0.43±0.28)〕比较,差异均有统计学意义(P<0.05);2、20、1 500 mg/L剂量组的DNA-蛋白质交联系数大于阴性对照组(P<0.05),且呈现剂量-效应关系。结论 NaNO2可诱导L929细胞出现毒物兴奋效应,并且在其剂量范围内可诱导细胞遗传物质的断裂性损伤。
Objective To study the effect of sodium nitrite (NaNO2) on the proliferation of L929 cells in vitro and to explore the damage of genetic material in this process. Methods L929 cells were cultured in vitro. The effects of different concentrations of NaNO2 on the proliferation of L929 cells were detected by tetrazolium bromide blue staining. Single cell gel electrophoresis (SDS-PAGE) and K-SDS precipitation assay Effect of NaNO2 on DNA damage and DNA-protein cross-linking in L929 cells. Results After treated with NaNO2 for 48h, the hormesis appeared in cell proliferation with the dose range of 0 ~ 0.0384mg / L. The tail length of L929 cells was (2.72 ± 0.99 and 3.26 ± 3.09 μm respectively, tail DNA% were (7.87 ± 6.63)% and (7.75 ± 4.90)%, respectively. Olive tail moments were (0.80 ± 0.64) and (0.79 ± 0.64) The difference between the two groups was statistically significant (P <0.05). Compared with the control group, the levels of DNA- The protein cross-linking coefficient was greater than that of the negative control group (P <0.05), and showed dose-effect relationship. Conclusion NaNO2 can induce the toxic effects of L929 cells and induce the damage of cytogenetic substances in the dose range.