γ-干扰素对培养人原代结膜及泪腺上皮细胞表达炎症相关因子的影响

Expression of inflammation-related cytokines in primary human conjunctival and lacrimal gland epithelial cells induced by 7-interferon

目的 了解γ-干扰素(γ-IFN)对体外原代培养的人结膜上皮细胞及泪腺上皮细胞的细胞间粘附分子(intercellular adhesion molecule-1,ICAM-1,CD54)、白细胞介素-1β(interl-eukin-1β,IL-1β)和白细胞介素-1受体1(interleukin-1 receptor1,IL-1R1)表达的影响,以初步探讨γ-IFN对人结膜及泪腺上皮细胞炎症损伤的作用。方法 混合消化液培养法获得人原代结膜上皮细胞,组织块培养法获得人泪腺上皮细胞,免疫组织化学方法鉴定。在细胞培养液中加入γ-IFN,浓度分别为0、30、100、300U·mL^(-1),37℃培养24h后收集细胞,应用流式细胞学及Western blot检测CD54的表达,Western blot检测IL-1β及IL-1R1的表达。结果 在γ-IFN浓度分别为0、30、100及300U·mL^(-1)条件下,结膜上皮细胞表达CD54的平均荧光强度为4.67、24.1、27.4及31.9;泪腺上皮细胞表达CD54的荧光强度分别为2.50、10.2、21.4及25.6。Western blotting的结果与流式细胞仪检测的结果相符,正常人结膜和泪腺上皮细胞有少量CD54表达,加入IFN-γ后CD54的表达增加,且呈剂量依赖性。Western blot的结果显示正常人结膜上皮细胞有少量IL-1R1表达,加入γ-IFN后IL-1β及IL-1R1的表达明显增加,且呈剂量依赖性,泪腺上皮细胞未检出IL-1β及IL-1R1的表达。结论 γ-IFN可?

Objective To evaluate the expression of intercellular adhesion molecule-1 (ICAM-1 ,CD54) , interleukin-1 p (IL-1 p) and interleukin-1 receptor 1 (IL-1R1)induced by y-interferon (y-IFN) in human conjunctival epithelial cells and lacrimal gland epithelial cells. Methods Human primary conjunctival epithelial cells and lacrimal gland epithelial cells were obtained from donor and identified by immunohistochemistry. These two kinds of cells were treated with different concentration of y-IFN(0,30,100 and 300U mL-1) for 24 hours. Then the expression of CD54 was detected by flow cytometry and western blot. Western blot was also used to evaluate the expression of IL-1 (3 and IL-1R1. Results Before treated with y-IFN, the mean intensity of CD54 expression was low in conjunctival and lacrimal gland epithelial cells(4. 67 and 2. 50, respectively) by flow cytometry. After treated with different concentration( 30, 100 and 300U-mL-1) of y-IFN, the mean intensity of CD54 expression increased significantly (conjunctival epithelial cells: 24.1,27. 4 and 31. 9; lacrimal gland epithelial cells: 10. 2,21. 4 and 25. 6). The result of western blotting also showed an increasing trend of the expression of C.D54 in these two kinds of cells after treatment of y-IFN. The expression of IL-1 p and IL-1R1 also increased greatly in conjunctival epithelial cells after treatment when using western blot. Conclusion y-IFN can induce the increasing expression of CD54 in human conjunctival and lacrimal gland epithelial cells, which suggests that the inflammatory injury caused by y-IFN in these two kinds of epithelium is partially mediated by CD54. y-IFN can also induce the increasing expression of IL-1 p and IL-1R1, which suggests that conjunctival epithelium can also become a resource of IL-1 in tear film under high concentration of y-IFN.