姜黄素对小鼠角质形成细胞核转录因子κB及其抑制因子IκBα的影响

Effects of Curcumin on the Expression of Nuclear Factor(NF)-κB and its Iinhibitor IκBα in Mice Keratinocytes

目的观察姜黄素对小鼠角质形成细胞291增殖的影响,并探讨姜黄素对核转录因子κB(NF-κB)及其抑制因子IκBα的干预作用。方法常规培养291细胞至90%融合,分别加入5,10,15,20,25μmol/L的姜黄素,继续培养3h和6h,MTT法检测不同浓度的姜黄素作用不同时间,291细胞增殖的情况;在常规培养至90%融合的291细胞中,分别加入肿瘤坏死因子(TNF-α)20ng/mL和/或姜黄素20μmol/L,继续培养3h,显微镜下观察细胞形态和活性;采用MTT法检测细胞吸光度;Western blot法分析各组细胞NF-κB及IκBα蛋白的表达情况。结果姜黄素对291细胞增殖抑制作用随剂量增大、时间延长而增加;显微镜下可见,姜黄素作用于291细胞3h后,细胞收缩,形态变圆,单个视野死亡细胞数较空白组和TNF-α组显著增加,姜黄素组细胞吸光度显著下降(P<0.05);Western blot实验结果显示,姜黄素组NF-κB蛋白表达显著下降,IκBα蛋白的表达显著升高(P<0.05);而TNF-α组NF-κB蛋白表达显著升高(P<0.05)。结论姜黄素抑制角质形成细胞增殖的机制,可能是抑制NF-κB表达,增加IκBα表达。

Objective To observe the effects of curcumin on the proliferation and expression of nuclear factor （NF） -κB and its inhibitor IκBα in mice keratinoeytes 291. Methods Cuhured 29t cells up to 90% confluence and then treated them with 5, 10, 15, 20, 25p, mol/L curcumin for 3h and 6h respectively, then optical density （OD） was measured in every group. Cultured 291 cells up to 90% cotffluence, added tumor necrosis factor α （TNF-α） （20ng/mL） and/or curcumin （20μmol/L） in every. well respectively and incubated for 3 hours. Cell morphological changes were observed by microscopy and cell viability was measured by using MTT assay, the expression of NF-κB and IκBα protein were detected by Western blot. Results MTT showed that the 291 cell proliferation was down regulated by curcumin significantly in a does and time de- pendent manner; after treated by TNF-α （20ng/mL） and/or curcumin （20μmol） for 3 hours, the 291 cells in curcumin group were contracted, turned round and there were more dead ceils than that in other groups. As assessed by MTF assay, curcumin could inhibite the proliferation of 291 cells （P 〈0.05）. Western blot a nalysis revealed in curcumin group NF-κBprotein decreased, IκBα protein significant increased （P 〈 0.05 ） ; but in TNF-α group NF-κB protein was increased （P 〈 0.05）. Conclusion Curcumin inhibits the proliferation of 291 cells significantly. The mechanism may be related with curcumin down regulating NF-κB but up regulating IκBα.