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目的研究特应血清被动致敏的人气道平滑肌细胞增殖、DNA合成、蛋白质合成等生物学特征。方法将正常人气道平滑肌细胞体外培养,用来源于2002~2003年在上海二医大附属瑞金医院就诊的哮喘患儿的特应血清(总IgE>106IU/L,特异性IgE≥Ⅲ级)对平滑肌细胞刺激、被动致敏,绘制被动致敏后的细胞生长曲线,比色法测定蛋白质合成量,3H-TdR结合试验测定DNA合成。结果特应血清被动致敏刺激的平滑肌细胞于培养的72h,96h,120h,细胞数分别为(5·21±0·87)×104/孔,(6·43±0·57)×104/孔,(7·25±0·65)×104/孔,同样条件下非特应血清刺激组细胞数分别为(3·78±0·50)×104/孔,(4·80±0·49)×104/孔,(5·55±0·77)×104/孔,两组在72h,96h,120h差异有显著性意义(P<0·01);特应血清致敏的平滑肌细胞蛋白质合成量为(3·81±0·52)mg/L,非致敏平滑肌细胞蛋白质合成量为(1·90±0·35)mg/L,两组相比差异有显著性(P<0·01),3H-TdR的结合力两组差异也有显著性意义[(196·4±45·2)cpm/孔和(127·2±33·3)cpm/孔,P<0·01]。结论特应血清致敏对平滑肌细胞分裂和蛋白质合成有显著促进作用,过度增殖的平滑肌细胞使气道平滑肌总量增加,是发生气道重建的重要结构基础。
Objective To study the biological characteristics of human airway smooth muscle cell proliferation, DNA synthesis and protein synthesis, which are passively sensitized by special serums. Methods Normal human airway smooth muscle cells were cultured in vitro. Serum from patients with asthma (total IgE> 106 IU / L, specific IgE ≧ III grade) from 2005 to 2003 in Ruijin Hospital of the Second Affiliated Hospital of Shanghai Medical College was used to detect smooth muscle cells Cell stimulation, passive sensitization, drawing passive cell sensitization of cell growth curve, colorimetric determination of protein synthesis, 3H-TdR binding assay DNA synthesis. Results The number of cells that were stimulated by passive sensitization of smooth muscle cells at 72h, 96h and 120h were (5 · 21 ± 0 · 87) × 104 / well, (6 · 43 ± 0 · 57) × 104 / (7.25 ± 0.65) × 104 / well. The number of cells in the non-specific serum-stimulated group under the same conditions were (3.78 ± 0.50) × 104 / well and (4.80 ± 0.49) ) × 104 / well and (5.55 ± 0.77) × 104 / well respectively. There was significant difference between the two groups at 72h, 96h and 120h (P <0.01). Serum sensitized smooth muscle cells The amount of protein synthesized in non-sensitized smooth muscle cells was (1.90 ± 0.35) mg / L, with a significant difference between the two groups (P <0. · 01). There was also a significant difference between the two groups in the binding capacity of 3H-TdR [(196.4 ± 45.2) cpm / well and (127.2 ± 33.3) cpm / well, P <0.01] . CONCLUSIONS: Serum sensitization can significantly promote smooth muscle cell division and protein synthesis. Over-proliferation of smooth muscle cells increases the total amount of airway smooth muscle, which is an important structural basis for airway remodeling.