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目的用蛋白酶和蛋白酶激活受体激动肽(PAR-AP)刺激A549细胞后检测细胞中白细胞介素IL-28和IL-29 mRNA表达情况以分析蛋白酶激活受体激活对人肺上皮细胞IL-28和IL-29基因表达的影响。方法用最佳工作浓度的凝血酶、胰蛋白酶、类胰蛋白酶及蛋白酶激活受体-1,2,3,4的激动肽刺激A549细胞后,分别在第2、8、16h收集细胞,用实时定量逆转录聚合酶链式反应(q-RT-PCR)方法分析A549细胞内IL-28和IL-29 mRNA表达。结果凝血酶作用A549后,细胞内IL-29 mRNA表达增强,高达对照组9.6倍;而细胞内IL-28 mRNA表达无明显变化。胰蛋白酶作用A549细胞后,细胞内IL-29和IL-28 mRNA表达增强,分别为对照组6.1倍和4.4倍。类胰蛋白酶作用A549细胞后,细胞内IL-29和IL-28 mRNA表达增强分别高达对照组的3.1倍和2.1倍。弹性蛋白酶作用A549细胞后,细胞内IL-29 mRNA表达增强高达对照组5.1倍而IL-28 mR-NA无明显变化。PAR-1AP(SFLLR)作用后,A549细胞内IL-29 mRNA表达增强为对照组1.7倍而IL-28 mRNA表达无明显变化。PAR-2AP(SLIGKV及tc-LIGRLO)作用后,A549细胞内IL-29和IL-28 mRNA表达增强分别为对照组4.9倍和2.4倍。PAR-3AP(TFRGAP)作用后,A549细胞内IL-29和IL-28 mRNA表达无明显变化。PAR-4AP(GYPGQV)作用后A549细胞内IL-29和IL-28 mRNA表达增强高达对照组4.1倍和5.5倍。结论 A549细胞蛋白酶激活受体的激活可以上调细胞内IL-29和IL-28 mRNA表达。
Objective To detect the expression of interleukin-28 (IL-28) and interleukin-29 (IL-29) mRNA in A549 cells stimulated with protease and protease activated receptor agonist peptide (PAR-AP) to analyze the effect of protease activated receptor on IL-28 And IL-29 gene expression. Methods A549 cells were stimulated with thrombin, trypsin, tryptase and protease activating receptor-1, 2, 3 and 4 at the optimal working concentrations. Cells were harvested at 2,8 and 16 h, respectively, Quantitative reverse transcription-polymerase chain reaction (q-RT-PCR) was used to analyze the mRNA expression of IL-28 and IL-29 in A549 cells. Results After the thrombin was treated with A549, the expression of IL-29 mRNA in the cells increased up to 9.6 times that of the control group, while the expression of IL-28 mRNA in the cells did not change significantly. After treated with trypsin, the expression of IL-29 and IL-28 mRNA in A549 cells increased by 6.1 and 4.4 times respectively. After tryptase A549 cells, the intracellular IL-29 and IL-28 mRNA expression increased 3.1 times and 2.1 times higher than that of the control group respectively. Elastase A549 cells, intracellular IL-29 mRNA expression increased up to 5.1 times the control group and IL-28 mR-NA no significant change. After treated with PAR-1AP (SFLLR), the expression of IL-29 mRNA in A549 cells increased 1.7-fold and the expression of IL-28 mRNA did not change significantly. After treated with PAR-2AP (SLIGKV and tc-LIGRLO), the mRNA expression of IL-29 and IL-28 in A549 cells increased by 4.9 and 2.4 times respectively. After treated with PAR-3AP (TFRGAP), the expression of IL-29 and IL-28 mRNA in A549 cells did not change significantly. The expression of IL-29 and IL-28 mRNA in A549 cells increased by 4.1-fold and 5.5-fold after treatment with PAR-4AP (GYPGQV). Conclusion Activation of A549 cell protease-activated receptor can up-regulate the expression of IL-29 and IL-28 mRNA.