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目的探讨姜黄素对IL-17诱导的人表皮角质形成细胞株(HaCaT细胞株)核转录因子NF-κB活化的影响,为其对某些炎症性皮肤病的治疗提供一定的理论依据。方法用IL-17刺激HaCaT细胞不同时间(0h,0.5h,1h,2h),或用姜黄素不同浓度(2.5μmol/L,5μmol/L,10μmol/L)预处理1h后,加入IL-17刺激2h,应用ELISA法结合捕获探针检测各组的NF-κBp65的DNA结合活性。提取IL-17刺激后不同时间点各组以及10μmol/L浓度姜黄素预处理组核蛋白,Westernblot方法检测各组NF-κBp65蛋白的表达。结果 IL-17能够有效地增加HaCaT细胞NF-κBp65的DNA结合活性及核蛋白表达。加入姜黄素共培养后,各浓度处理组的NF-κBp65DNA结合活性均下降(P<0.01),10μmol/L姜黄素处理引起NF-κBp65的蛋白表达水平明显下降(P<0.01)。结论姜黄素能够有效抑制IL-17诱导的HaCaT细胞NF-κBp65的DNA结合活性以及核蛋白的表达,从而抑制NF-κB的活化。
Objective To investigate the effect of curcumin on the activation of NF-κB induced by IL-17 in human epidermal keratinocyte (HaCaT) cell line and to provide some theoretical evidences for the treatment of some inflammatory skin diseases. Methods HaCaT cells were stimulated with IL-17 at different times (0h, 0.5h, 1h, 2h) or pretreated with different concentrations of curcumin (2.5μmol / L, 5μmol / L, 10μmol / L) After stimulated for 2 hours, the DNA binding activity of NF-κBp65 in each group was detected by ELISA and capture probe. At different time points after stimulation with IL-17, the nuclear proteins of curcumin pretreatment group and 10μg / L curcumin were extracted. The expression of NF-κB p65 protein in each group was detected by Western blot. Results IL-17 could effectively increase the DNA-binding activity and nuclear protein expression of NF-κBp65 in HaCaT cells. After co-cultured with curcumin, the NF-κBp65 DNA binding activity of all the treated groups decreased (P <0.01). The protein expression of NF-κBp65 was significantly decreased after treated with 10μmol / L curcumin (P <0.01). Conclusion Curcumin can effectively inhibit the DNA-binding activity and nuclear protein expression of NF-κBp65 in HaCaT cells induced by IL-17, and thus inhibit the activation of NF-κB.