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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 effect of curcumin on the proliferation of mouse keratinocytes 291 and to explore the intervention of curcumin on nuclear factor κB (NF-κB) and its inhibitory factor IκBα. Methods 291 cells were routinely cultured to 90% confluence. Curcumin at 5, 10, 15, 20 and 25μmol / L was added respectively and the cells were cultured for 3h and 6h. MTT assay was used to detect the proliferation of 291 cells treated with different concentrations of curcumin The cells were cultured in 90% confluent 291 cells with 20ng / mL tumor necrosis factor (TNF-α) and 20μmol / L curcumin respectively for 3h. The cell morphology and activity were observed under MTT assay The cell absorbance was detected by Western blot. The expression of NF-κB and IκBα in each group was analyzed by Western blot. Results Curcumin inhibited the proliferation of 291 cells with increasing dose and prolonged time. Under the microscope, after curcumin treatment on 291 cells for 3 h, the cells contracted and became round in morphology. The number of dead cells in a single field of vision was significantly higher than that of blank group and TNF- (P <0.05). The results of Western blot showed that the expression of NF-κB protein in curcumin group was significantly decreased and the expression of IκBα protein was significantly increased (P <0.05), while TNF -α group of NF-κB protein expression was significantly increased (P <0.05). Conclusion Curcumin may inhibit the proliferation of keratinocytes by inhibiting the expression of NF-κB and increasing the expression of IκBα.