白细胞介素22对人表皮角质形成细胞CC趋化因子配体27表达的影响

Effects of interleukin- 22 on the expression of CC chemokine ligand 27 in human epidermal keratinocytes

目的 研究白细胞介素22（IL-22）对人表皮角质形成细胞CC趋化因子配体27（CCL27）表达的影响及机制。方法 免疫组化法检测10例银屑病患者皮损及5例健康对照组皮肤中CCL27的表达。培养HaCaT细胞，分为8组：对照组用PBS处理，IL-22处理组分别用12.5、25、50、100、200 μg/L IL-22处理，信号通路阻断组先分别加入50 μmol/L AG490（JAK2/STAT3通路抑制剂）或PD98059（MAPK-ERK1/2通路抑制剂）阻断处理，2 h后各加入50 μg/L IL-22，继续于37 ℃、5% CO2孵箱中培养。孵育24 h 后，分别提取 HaCaT 细胞总蛋白并收集上清液，Western印迹、ELISA法分别检测CCL27表达。结果 免疫组化检测示，银屑病患者皮损中CCL27的表达水平明显高于正常人皮肤。Western 印迹显示，HaCaT细胞分别用12.5、25、50、100、200 μg/L IL-22处理后，CCL27相对表达量逐渐升高至0.491 ± 0.013、0.620 ± 0.019、0.623 ± 0.014、0.802 ± 0.052、1.138 ± 0.013，均较对照组（0.413 ± 0.013）升高（P＜0.01）；IL-22 ＋ AG490组及IL-22 ＋ PD98059组CCL27表达量分别为0.411 ± 0.019和0.280 ± 0.012，均低于50 μg/L IL-22组（均P＜0.01）。ELISA法检测显示，各组HaCaT细胞CCL27分泌水平变化趋势与Western印迹法检测结果一致。结论 IL-22可促进HaCaT 细胞CCL27表达，其机制可能与MAPK-ERK1/2和JAK2/STAT3通路有关。

Objective To evaluate the effects of interleukin-22 （IL-22） on the of CC chemokine ligand 27 （CCL27） in human epidermal keratinocytes, and to explore its mechanism. Methods Immunohistochemical study was performed to determine the of CCL27 in skin lesions of 10 patients with psoriasis and skin tissues of 5 healthy controls. Cultured HaCaT cells were divided into 8 groups： control group treated with PBS, 5 IL-22 groups treated with 12.5, 25, 50, 100 and 200 μg/L IL-22 respectively, 2 signaling pathway inhibition groups treated with 50 μmol/L AG490 （JAK2/STAT3 signaling pathway inhibitor） or PD98059 （MAPK-ERK1/2 signaling pathway inhibitor） for 2 hours followed by the treatment with 50 μg/L IL-22 in the 5% CO2 incubator at 37 ℃. After 24-hour cultivation, total proteins were extracted, and culture supernatants were collected, and both Western blot analysis and enzyme-linked immunosorbent assay（ELISA）were performed to determine the of CCL27. Results Immunohistochemical study showed that the of CCL27 was significantly higher in the skin lesions of the patients with psoriasis than in the skin tissues of the healthy controls. Western blot analysis revealed that the protein of CCL27 in the 12.5-, 25-, 50-, 100- and 200-μg/L IL-22 groups was 0.491 ± 0.013, 0.620 ± 0.019, 0.623 ± 0.014, 0.802 ± 0.052 and 1.138 ± 0.013 respectively, which were all higher than that in the control group （0.413 ± 0.013, all P 〈 0.01）. The of CCL27 was significantly lower in the IL-22 ＋ AG490 group （0.411 ± 0.019） and IL-22 ＋ PD98059 group （0.280 ± 0.012） than in the 50-μg/L IL-22 group （both P 〈 0.01）. ELISA also showed the same trend of changes in the level of CCL27 in the above groups as Western blot showed. Conclusion IL-22 can promote the of CCL27 in HaCaT cells, which may be associated with the MAPK-ERK1/2 and JAK2/STAT3 signaling pathways.