Notch信号通路与银屑病发病的关系及作用机制

发布时间：2019-04-07 17:32

【摘要】：目的探讨Notch信号通路与银屑病发病的关系及作用机制。方法分别用RT-PCR、Western blotting技术检测30例银屑病患者皮损区皮肤组织(病例组)及其周边非皮损区皮肤组织(对照组)、30例正常人皮肤组织(正常组)中Notch1、Jagged1 mRNA和蛋白表达;将人永生化角质形成细胞Ha Ca T随机分为Jagged1-Fc融合蛋白组、DAPT组、常规对照组,分别加入含Notch信号通路特异性激活剂(Jagged1-Fc融合蛋白,终浓度0、0.5、1、2μg/m L)、Notch信号通路特异性阻断剂(DAPT,终浓度0、5、10、20μmol/L)、常规细胞培养液干预24、48、72 h。用Western blotting法检测细胞内Bcl-2、Bax、Keratin1、Involucrin蛋白表达,用MTT法检测Ha Ca T细胞增殖活力,用流式细胞术检测细胞凋亡率。结果病例组Notch1、Jagged1 mRNA及蛋白相对表达量高于对照组与正常组(P均0.05)。干预0、24、48、72 h,Jagged1-Fc融合蛋白组细胞增殖率随干预浓度升高依次升高,DAPT组随干预浓度降低依次降低(P均0.05)。与常规对照组比较,Jagged1-Fc融合蛋白组细胞凋亡率降低、DAPT组升高(P均0.05)。与常规对照组比较,Jagged1-Fc融合蛋白组Bcl-2蛋白相对表达量升高,Bax蛋白相对表达量降低(P均0.05);DAPT组Bcl-2蛋白相对表达量降低,Bax蛋白相对表达量升高(P均0.05)。与常规对照组比较,Jagged1-Fc融合蛋白组Keratin1及Involucrin蛋白相对表达量降低(P均0.05);DAPT组Keratin1及Involucrin蛋白相对表达量升高(P均0.05)。结论银屑病的发生与Notch信号通路的激活相关,其可能的作用机制为Notch信号通路促进角质形成细胞增殖、抑制其凋亡与分化。
[Abstract]:Objective to investigate the relationship between Notch signaling pathway and the pathogenesis of psoriasis. Methods RT-PCR,Western blotting technique was used to detect the expression of Notch1,Jagged1 mRNA and protein in the skin tissue of 30 patients with psoriasis and its surrounding non-skin tissue (control group), and in 30 normal skin tissues (normal group). Human immortalized keratinocytes (Ha Ca T) were randomly divided into three groups: Jagged1-Fc fusion protein group, DAPT group and routine control group. Jagged1-Fc fusion protein containing Notch signaling pathway specific activator (Jagged1-Fc fusion protein, final concentration 0, 0.5, 1, 2 渭 g / m L),) was added respectively. Notch signaling pathway specific blockers (DAPT, final concentration 0, 5, 10, 20 渭 mol / L),) were treated with 24, 48, 72 h. Western blotting assay was used to detect the expression of Bcl-2,Bax,Keratin1,Involucrin protein, MTT method was used to detect the proliferation activity of Ha Ca T cells, and flow cytometry was used to detect the apoptosis rate. Results the relative expression of Notch1,Jagged1 mRNA and protein in the case group was higher than that in the control group and the normal group (P 0.05). At 0,24,48,72 h after intervention, the proliferation rate of Jagged1 / FC fusion protein group increased with the increase of the intervention concentration, while that of the DAPT group decreased with the decrease of the intervention concentration (P all 0.05). Compared with the normal control group, the apoptosis rate of Jagged1-Fc fusion protein group was lower and that of DAPT group was higher (all P 0.05). Compared with the normal control group, the relative expression of Bcl-2 protein in Jagged1-Fc fusion protein group was higher than that in control group, while the relative expression of Bax protein was decreased (all P 0.05). The relative expression of Bcl-2 protein was decreased and the relative expression of Bax protein was increased in DAPT group (all P 0.05). Compared with the normal control group, the relative expression of Keratin1 and Involucrin protein in Jagged1-Fc fusion protein group was lower than that in control group (P < 0.05). The relative expression of Keratin1 and Involucrin protein in); DAPT group was significantly higher than that in control group (P < 0.05). Conclusion the pathogenesis of psoriasis is related to the activation of Notch signaling pathway. Its possible mechanism is that Notch signaling pathway promotes the proliferation of keratinocytes and inhibits the apoptosis and differentiation of keratinocytes.
【作者单位】：山东中医药大学;
【分类号】：R758.63

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