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目的比较纳米银及其释放的银离子对人永生化角质形成(Ha Ca T)细胞炎性因子分泌的影响,并进一步探讨纳米银的吸附作用对炎性因子测定的影响。方法采用不同浓度[0(对照)~100μg/ml]的纳米银染毒Ha Ca T细胞24 h,采用乳酸脱氢酶(LDH)释放试验测定纳米银的细胞毒性;选择0(对照)、2.5、5、10、20和40μg/ml的纳米银或其释放的银离子[0(对照)、0.05、0.1、0.2、0.4和0.8μg/ml]染毒Ha Ca T细胞24 h,采用酶联免疫吸附试验(ELISA)检测细胞培养液中的白细胞介素(IL)-6和IL-8的含量。在无细胞体系中,将终浓度为0(对照)、5、40和100μg/ml的纳米银与IL-6或IL-8标准品溶液混匀静置6 h后离心去除纳米银颗粒,采用ELISA测定上清液中IL-6、IL-8的含量。结果纳米银浓度低于75μg/ml时无明显细胞毒性。与对照组比较,2.5、5、20μg/ml纳米银染毒Ha Ca T细胞IL-6的分泌量和5、10、20μg/ml纳米银染毒Ha Ca T细胞IL-8的分泌量均较高,差异均有统计学意义(P<0.01)。与对照组比较,仅0.1、0.8μg/ml银离子染毒Ha Ca T细胞IL-8的分泌量较高,差异均有统计学意义(P<0.05);而各剂量银离子染毒Ha Ca T细胞IL-6的分泌量均无明显改变。在无细胞体系中,与对照组比较,40、100μg/ml纳米银暴露后体系中IL-6的含量和100μg/ml纳米银暴露后体系中IL-8的含量均较低,差异均有统计学意义(P<0.01,P<0.05)。结论本实验条件下低浓度纳米银可诱导Ha Ca T细胞IL-6和IL-8的分泌量增加,其对IL-6分泌量的影响可能与纳米银颗粒密切相关,而对IL-8分泌量的影响可能与悬液中释放的银离子有关;高浓度纳米银对IL-6和IL-8具有吸附性。
Objective To compare the effects of silver nanoparticles and silver ions released on the secretion of inflammatory cytokines in human immortalized human keratinocytes (HaCa T) cells, and to explore the effect of nano-silver adsorption on the determination of inflammatory cytokines. Methods HaCa T cells were exposed to different concentrations of [0 (control) ~ 100 μg / ml] for 24 h, and the cytotoxicity of nano-silver was determined by lactate dehydrogenase (LDH) release assay. HaCa T cells were treated with 5, 10, 20 and 40 μg / ml silver nanoparticles or their released silver ions [0 (control), 0.05, 0.1, 0.2, 0.4 and 0.8 μg / ml] Immunosorbent assay (ELISA) was used to detect the levels of interleukin (IL) -6 and IL-8 in the cell culture medium. In the cell-free system, the final concentration of 0 (control), 5,40 and 100μg / ml of silver nanoparticles and IL-6 or IL-8 standard solution mixed 6 hours after centrifugation to remove the nano-silver particles, using The contents of IL-6 and IL-8 in the supernatant were measured by ELISA. Results Nano-silver concentration of less than 75μg / ml no significant cytotoxicity. Compared with the control group, the secretions of IL-6 secreted by 2.5, 5, 20μg / ml Nano-Ag in HaCa T cells and the secretion of IL-8 in 5,10,20μg / ml Nano-Ag HaCa T cells High, the difference was statistically significant (P <0.01). Compared with the control group, the secretion of IL-8 in HaCa T cells exposed to 0.1, 0.8 μg / ml silver ions was significantly higher than that of the control group (P <0.05) T cells IL-6 secretion were no significant change. In the cell-free system, compared with the control group, the contents of IL-6 in the system after exposure to 40,100μg / ml nano-silver and the content of IL-8 in the system after 100μg / ml nano-silver exposure were all lower Significance (P <0.01, P <0.05). Conclusions Under the experimental conditions, the secretion of IL-6 and IL-8 by HaCa T cells with low concentration of nano-silver can be increased, and the effect on the secretion of IL-6 may be closely related to the nano-silver particles, while the secretion of IL-8 The effect of the amount may be related to the release of silver ions in the suspension; the high concentration of silver nanoparticles is adsorptive to IL-6 and IL-8.