中文摘要：

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。

英文摘要：

Objective To investigate the proinflammatory effects of silver nanoparticles（Ag NPs） and silver ions（Ag＋） on human skin keratinocytes（Ha Ca T） and the possible effects of Ag NPs on adsorbing proinflammatory cytokines. Methods Ha Ca T cells were treated with Ag NPs at the doses of 0,0.5,1,5,25,50,75 and 100 μg/ml respectively,for 24 h and cell viability was assessed by lactate dehydrogenase（LDH） release assay. Ag NPs and Ag＋-dependent effects on interleukin（IL）-6 and IL-8 release were measured by enzyme-linked immunosorbent assay（ELISA）. In cell-free system,Ag NPs were incubated with IL-6 or IL-8protein standard solution for 6 h at different concentrations（0,5,40 and 100 μg/ml） and cytokine adsorption capacity of Ag NPs was also evaluated by ELISA. Results Ag NPs with concentrations less than 75 μg/ml did not cause cytotoxicity. At low concentrations（2.5,5 and 20 μg/ml）,a significant increase of IL-6 level was observed with Ag NPs. A dramatic increase of IL-8after exposure to 5-20 μg/ml of Ag NPs and 0.1,0.8 μg/ml of Ag＋were also found. In the cell-free system,IL-6 level was significantly reduced at Ag NPs doses of 40 and 100 μg/ml,whereas IL-8 level was reduced significantly only in the presence of 100 μg/ml Ag NPs. Conclusion Ag NPs may trigger proinflammation in Ha Ca T cells at lower dose and both Ag NPs and Ag＋may play a considerable role. Ag NPs can adsorb IL-6 and IL-8 at higher dose.