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目的探讨核因子κB(NFκB)在体外培养的人视网膜色素上皮(human retinal pigment epithelium,hRPE)细胞中的基础表达以及吡咯二硫代氨基甲酸乙酯(pyrolli dinedi thio carbamate,PDTC)、IL1β对NFκB表达的影响。方法体外培养的hRPE细胞经同步化后分2组分别加药:(1)无PDTC组:分别加入IL1β、生理盐水(用于检测NFκB在hRPE中的基础表达);(2)PDTC组:分别加入IL1β、生理盐水(用于检测NFκB在PDTC预处理后hRPE中的表达)。免疫荧光抗体染色、流式细胞计数法(flow cyto metry,FCM)测定上述2组标本中hRPE的NFκB的表达率。结果NFκB在hRPE中的基础表达率为8.05%,IL1β作用后表达率升高到30.26%;hRPE细胞经PDTC预处理后,NFκB的表达率下降为3.74%,加入IL1β(10μg·L-1)作用后表达率为3.66%.结论IL1β可以显著提高NFκB在hRPE细胞中的表达;PDTC可以显著降低体外培养的hRPE中的NFκB表达率,PDTC亦可显著抑制由IL1β诱导的NFκB的活化过程。
Objective To investigate the basic expression of nuclear factor κB (NFκB) in cultured human retinal pigment epithelium (hRPE) cells and the effect of IL1β on NFκB The impact of expression. Methods The hRPE cells cultured in vitro were divided into two groups after dosing: (1) without PDTC group: IL1β and normal saline (for detecting the basic expression of NFκB in hRPE); (2) PDTC group: IL1β, physiological saline (for detection of NFκB expression in hRPE after pretreatment of PDTC) was added. Immunofluorescence staining and flow cytometry (FCM) were used to detect the expression of NFκB in hRPE. Results The basal expression of NFκB in hRPE was 8.05%, and the expression of NFκB was up to 30.26% after treated with IL1β. The expression of NFκB was down-regulated to 3.74% in hRPE cells after treatment with PDTC. IL1β (10μg · L-1) After treatment, the expression rate was 3.66% .Conclusion IL1β can significantly increase the expression of NFκB in hRPE cells; PDTC can significantly reduce the expression of NFκB in hRPE cultured in vitro, and PDTC can significantly inhibit the activation of NFκB induced by IL1β.