目的探讨CLP-19对RAW264.7细胞的免疫调节及其在炎症中的作用。方法以50μg/ml的CLP-19作用于RAW264.7细胞后不同时间(0.5、1、2.5、5、10和20 h),及不同质量浓度(1、10、50、100和200μg/ml)的CLP-19作用于RAW264.7细胞20 h,观察细胞TNF-α、IL-10、MCP-1、ICAM-1的mRNA水平的变化情况。另用50μg/ml的CLP-19预处理细胞5、10、和20 h后,再以LPS刺激,4 h后提取细胞RNA,观察上述细胞因子的变化,以CLP-19与LPS预混组为对照。结果 CLP-19单独作用于RAW264.7细胞后,CLP-19对MCP-1的mRNA调节呈显著的时间依赖性和剂量依赖性,高剂量的CLP-19可上调ICAM-1、TNF-α和IL-10的mRNA水平。以CLP-19预处理RAW264.7细胞不同时间(5、10和20 h)后,均可显著抑制LPS所刺激的炎症因子上调,各预处理组间无统计学差异。结论 CLP-19能改变细胞的免疫状态,并抑制LPS诱导的免疫反应。
In order to investigate the immunoregulatory effects of CLP-19, RAW264.7 cells were treated with the peptide CLP-19 (50 μg/ml) for different times (0.5, 1, 2.5, 5, 10, 20 h), and with different concentrations ofCLP-19 (1, 10, 50, 100, 200 μg/ml) for 20 hours, respectively. Moreover, RAW264.7 cells were pretreated with CLP-19 for different times (5, 10, 20 h), then treated with LPS for 4 h. The total RNAs were extracted and themRNA expressions of TNF-α, MCP-1, IL-10, and ICAM-I were measured by RT-PCR. Results showed that CLP- 19 significantly upregulated the mRNA expression of MCP-1 in RAW264.7 cells in dose-dependent and time-dependent manner, while high concentration of CLP-19 could upregulate the mRNA expression of ICAM-1, TNF- α, and IL-10 in RAW264.7 cells. Furthermore, the pretreatment of CLP-19 significantly inhibited upregulation ofinflammatory cytokines result from LPS treatment. In conclusion, CLP-19 can regulate the immune status of cells and suppress LPS-indueed immune response.