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目的观测自行研制的脂氧化酶抑制剂诺帝(Nordy)对人恶性胶质瘤细胞系 U87中甲酰化肽受体(FPR)的表达和激活后功能活性的影响。方法采用间接免疫荧光标记、激光共聚焦扫描显微术观测诺帝对 U87系人恶性胶质瘤细胞 FPR 表达的影响;FPR 激动剂 N-甲酰化的甲硫酰-亮氨酸-苯丙氨酰胺(fMLF)激活 FPR 后加入诺帝,实验分为以下3组:①对照组,②fMLF 刺激组,③诺帝处理组,分别采用四甲基偶氮唑盐(MTY)法、逆转录聚合酶链式反应(RT-PCR)和双抗夹心酶联免疫吸附法(ELISA)检测诺帝对 FPR 活化后引起的细胞增殖及细胞分泌血管内皮生长因子(VEGF)和白细胞介素8(IL-8)的作用。结果诺帝(100 μmol/L)明显抑制 U87细胞 FPR 受体的表达,同时对FPR 激动剂 fMLF(100 nmol/L)诱导的 U87细胞增殖和血管生成因子 VEGF、IL-8 mRNA 表达和蛋白的分泌具有明显的抑制作用。对照组 U87细胞分泌一定量的 VEGF 和 IL-8,分别为(4.14±0.28)ng/ml 和(4.03±0.59)ng/ml;fMLF 作用于 U87细胞36 h 后,VEGF 和 IL-8蛋白水平均显著高于对照组(P<0.05),分别为(6.46±0.33)ng/ml 和(7.54±0.73)ng/ml;诺帝作用于 U87细胞12 h后加入 fMLF 共同处理36 h 后,VEGF 和 IL-8蛋白水平分别为(3.59±0.33)ng/ml 和(3.13±0.48)ng/ml,均显著低于对照组(P<0.05)。结论诺帝对人恶性胶质瘤细胞的 FPR 表达和该受体活化后的促瘤细胞增殖及促血管生成因子 VEGF、IL-8 mRNA 表达和蛋白的分泌具有抑制作用,提示诺帝具有抑制肿瘤生长和抗血管生成的作用。
Objective To observe the effect of self-developed lipoxygenase inhibitor Nordy on the expression and activation of formin peptide receptor (FPR) in human malignant glioma cell line U87. Methods Indirect immunofluorescence labeling and confocal laser scanning microscopy were used to observe the effect of Nordy on FPR expression in U87 human glioblastoma cells. FPR agonist N-formylated methionyl-leucine-phenylpropion (FMLF), and then add Nordy to FPR. The experiment is divided into the following three groups: ① control group, ②fMLF stimulation group and ③ Nordy treatment group, MTT method and reverse transcription polymerization The effect of NOD on cell proliferation and the secretion of vascular endothelial growth factor (VEGF) and interleukin-8 (IL-8) induced by FPR activation were detected by RT-PCR and ELISA. 8) role. RESULTS: Nordy (100 μmol / L) significantly inhibited the expression of FPR receptor in U87 cells. The effect of fPRF (100 nmol / L) on the proliferation of U87 cells and the expressions of VEGF, IL-8 mRNA and protein Secretion has a significant inhibitory effect. U87 cells in control group secreted a certain amount of VEGF and IL-8, which were (4.14 ± 0.28) ng / ml and (4.03 ± 0.59) ng / ml, respectively. (6.46 ± 0.33) ng / ml and (7.54 ± 0.73) ng / ml respectively when compared with the control group (P <0.05). When the Nordy treated U87 cells for 12 hours and then treated with fMLF for 36 hours, VEGF (3.59 ± 0.33) ng / ml and (3.13 ± 0.48) ng / ml, respectively, were significantly lower than those in the control group (P <0.05). CONCLUSION: Nordy inhibits the expression of FPR on human glioblastoma cells and promotes the proliferation of tumor-promoting cells and the expression of angiogenic factors VEGF, IL-8 mRNA and protein, suggesting that Nordy has the ability of inhibiting tumor Growth and anti-angiogenic effects.