李良成, 侯琦, 程桂芳, 张成义. 大鼠中性粒细胞与滑膜细胞粘附模型的建立J. 药学学报, 2000, 35(2): 99-102.
引用本文: 李良成, 侯琦, 程桂芳, 张成义. 大鼠中性粒细胞与滑膜细胞粘附模型的建立J. 药学学报, 2000, 35(2): 99-102.
Li Liangcheng, Cheng Guifang, Zhang Chengyi, . ESTABLISHMENT OF A MODEL FOR ADHESION OF POLYMORPHONUCLEAR LEUKOCYTE TO SYNOVIAL CELLJ. Acta Pharmaceutica Sinica, 2000, 35(2): 99-102.
Citation: Li Liangcheng, Cheng Guifang, Zhang Chengyi, . ESTABLISHMENT OF A MODEL FOR ADHESION OF POLYMORPHONUCLEAR LEUKOCYTE TO SYNOVIAL CELLJ. Acta Pharmaceutica Sinica, 2000, 35(2): 99-102.

大鼠中性粒细胞与滑膜细胞粘附模型的建立

ESTABLISHMENT OF A MODEL FOR ADHESION OF POLYMORPHONUCLEAR LEUKOCYTE TO SYNOVIAL CELL

  • 摘要: 目的:建立大鼠中性粒细胞(PMN)与滑膜细胞(RSC)的粘附模型。方法:MTT比色法。结果:500 U.mL-1 rhTNF-α与IL-1刺激RSC 12 h对PMN有明显的粘附作用, MTT-PMN细胞数与540 nm吸光度值有良好的线形相关性;氢化考的松(hydrocortisone, Hy)和美洛昔康(meloxicam, Melo)均可抑制PMN与500 U.mL-1 rhTNF-α/或IL-1诱导的RSC粘附,提示Hy和Melo治疗类风湿性关节炎(RA)可能与其抑制PMN与RSC粘附作用有关。结论:以MTT比色法为基础建立的检测大鼠PMN与RSC的粘附模型,可用于PMN与RSC粘附抑制剂的筛选。

     

    Abstract: AIM: To establish a new and simple method of quantitating adhesion of neutrophil or polymorphonuclear leukocyte(PMN) to rat synovial cell(RSC) using MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphemyl tetrazolium bromide]-stained PMN cell. METHODS: PMN were incubated with MTT, and formed MTT labeled PMN(MTT-PMN) with a blue dye of formazan. After incubation of the MTT-PMN and RSC, non-adhered cells were washed out. Adhered cells were lysed with dimethylsulfoxide and quantitated by measure the absorbance at 540 nm. The concentration and time courses of rhTNF-α and IL-1 which stimulate RSC were studied. According to the positive-cross test, moreover, it were also studied that the inhibitory effects of hydrocortisone (Hy) and meloxicam (Melo) on the adhesion of PMN to RSC which were stimulated by rhTNF-α and IL-1 at 500 U.mL-1 for 12 h. RESULTS: The absorbance at 540 nm was shown to be well correlated with the cell numbers of MTT-PMN(0.25~2×109 cell.L-1) adhesion to RSC which were stimulated by 500 U.mL-1 rhTNF-α and IL-1 for 12 h. Hydrocortisone inhibited the adhesion of PMN to RSC stimulated by 500 U.mL-1 rhTNF-α and IL-1, the IC50 was 2.6×10-7 mol.L-1, 2.29×10-9 mol.L-1, respectively. But IC50 of meloxicam were 6.1×10-7 mol.L-1 and 1.25×10-6 mol.L-1, respectively. CONCLUSION: A new and simple method of quantitating adhesion of PMN to RSC was established, by using MTT-stained PMN cell. This method can be used for screening inhibitors of PMN adhesion to RSC.

     

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