Ouyang Ping, Peng Li-Sheng, Yang Hong, Peng Wen-Lie, Wu Wen-Yan, Xu An-Long
Department of Biochemistry, School of Life Science, Sun Yat-Sen University, Guangzhou, PR China.
Sheng Li Xue Bao. 2003 Apr 25;55(2):128-34.
The purposes of this study was to determine the effects of recombinant human interleukin-10 (rhIL-10) on proliferation of vascular smooth muscle cells (VSMCs) stimulated by advanced glycation end products (AGE) and neointima hyperplasia after rat carotid arterial injury. Rat aortic VSMCs were cultured and treated with rhIL-10 or AGE respectively, and then co-treated with rhIL-10 and AGE. Proliferation of VSMCs was quantified by colormetric assay. Cell cycle analysis was performed by flow cytomertry. Sprague-Dawley rats were treated with recombinant human IL-10 (rhIL-10) for 3 d after carotid arteries injury. The ratio of neointima to media area at the site of arterial injury was measured 28 d after balloon injury. The p44/42 MAPK activity was evaluated by the immunoblotting technique using anti-p44/42 phospho-MAPK antibody. Compared to control, AGE stimulated VSMCs proliferation. rhIL-10 alone had no effect on VSMCs growth. With AGE stimulation, rhIL-10, at dose as low as 10 ng/ml, inhibited VSMCs growth (P<0.05). The cell number in G(0)/G(1) phase of AGE and rhIL-10 co-treatment group was higher than that of AGE treatment alone (P<0.01) by flow cytometry analysis. Compared with the control group of neointima hyperplasia in rats, the ratio of neointima to media area of recombinant human IL-10 group was reduced by 45% (P<0.01). The p44/42 MAPK activity was significantly enhanced by AGE. The AGE effects were opposed by rhIL-10. The anti-inflammatory cytokine rhIL-10 inhibits AGE-induced VSMCs proliferation. Recombinant human IL-10 also inhibited neointima hyperplasia after carotid artery injury in rats. The results suggest the possibility that recombinant human IL-10, as a potential therapeutic approach, prevents neointimal hyperplasia.
本研究旨在确定重组人白细胞介素-10（rhIL-10）对晚期糖基化终产物（AGE）刺激的血管平滑肌细胞（VSMC）增殖以及大鼠颈动脉损伤后新生内膜增生的影响。培养大鼠主动脉VSMC，分别用rhIL-10或AGE处理，然后用rhIL-10和AGE共同处理。通过比色法对VSMC的增殖进行定量。通过流式细胞术进行细胞周期分析。对Sprague-Dawley大鼠颈动脉损伤后用重组人IL-10（rhIL-10）处理3天。球囊损伤28天后测量动脉损伤部位新生内膜与中膜面积之比。使用抗p44/42磷酸化丝裂原活化蛋白激酶（MAPK）抗体通过免疫印迹技术评估p44/42 MAPK活性。与对照组相比，AGE刺激VSMC增殖。单独的rhIL-10对VSMC生长无影响。在AGE刺激下，低至10 ng/ml剂量的rhIL-10抑制VSMC生长（P<0.05）。通过流式细胞术分析，AGE与rhIL-10共同处理组G(0)/G(1)期的细胞数量高于单独AGE处理组（P<0.01）。与大鼠新生内膜增生对照组相比，重组人IL-10组新生内膜与中膜面积之比降低了45%（P<0.01）。AGE显著增强p44/42 MAPK活性。rhIL-10可对抗AGE的作用。抗炎细胞因子rhIL-10抑制AGE诱导的VSMC增殖。重组人IL-10还可抑制大鼠颈动脉损伤后的新生内膜增生。结果提示重组人IL-10作为一种潜在治疗方法预防新生内膜增生的可能性。
