重组人白介素-10对TNF-α诱导的血管外膜成纤维细胞syndecan-4蛋白表达的影响

Recombinant human interleukin-10 inhibits expression of syndecan-4 protein in adventitial fibroblasts induced by tumor necrosis factor-α in vitro

目的观察重组人白介素-10(IL-10)对肿瘤坏死因子-α(TNF-α)刺激的血管外膜成纤维细胞(NIH/3T3细胞)增殖和syndecan-4蛋白表达的影响。方法采用体外培养的NIH/3T3细胞,分别应用终质量浓度为20 ng/mL TNF-α、100 ng/mL IL-10、200 ng/mL IL-10、100 ng/mL IL-10联用20 ng/mL TNF-α、200 ng/mL IL-10联用20 ng/mL TNF-α作用24 h,并设对照组进行比较,采用MTS/PMS法确定NIH/3T3细胞的增殖状态,利用Western blot蛋白免疫印迹法测定NIH/3T3细胞中syndecan-4蛋白的表达。结果(1)MTS/PMS法测定各组细胞增殖率统计分析显示:与对照组比较,TNF-α组能显著刺激NIH/3T3细胞的增殖(P<0.001),而IL-10不同剂量组单独应用对NIH/3T3细胞的增殖均无明显作用(P>0.05)。IL-10联合TNF-α共同作用组与TNF-α组比较,NIH/3T3细胞增殖均显著减少(P<0.01)。(2)Westernblot蛋白免疫印迹法测定显示:与对照组比较,TNF-α组能显著刺激NIH/3T3细胞中的syndecan-4蛋白表达(P<0.05),而IL-10不同剂量组单独应用对NIH/3T3细胞的syndecan-4蛋白表达均无明显影响(P>0.05)。IL-10联合TNF-α共同作用组与TNF-α组比较,NIH/3T3细胞的syndecan-4蛋白表达显著降低(P<0.001)。结论IL-10可明显抑制TNF-α诱导的NIH/3T3细胞增殖及细胞syndecan-4蛋白的表达。

Objective To investigate the effect of recombinant human interleukin-10（IL-10） on proliferation of adventitial fibroblasts and expression of syndeean-4 protein induced by tumor necrosis factor-α （ TNF-α）. Methods NIH 3T3 cells were cultured in vitro and exposed to 20 ng/mL TNF-α,100 ng/mL IL-10, 200 ng/mL IL-10,100 ng/mL IL-10 with 20 ng/mL TNF-α,and 200 ng/mL IL-10 with 20 ng/mL TNF-α respectively. All groups were treated for 24 h in vitro, and were compared with the control group. The ratio of proliferation of NIH/3T3 cells was determined by non-radioactive MTS/PMS assay and the expression of syndecan-4 protein was evaluated by Western blotting using anti-syndecan-4 antibodies. Results Statistical analysis showed that, （1） Compared to the control group,TNF-α significantly stimulated the proliferation of NIH/3T3 cells at the concentration of 20 ng/mL in TNF-α group （ P 〈 0. 001 ）. IL-10 alone had no effect on NIH/3T3 cells growth （P 〉 0.05 ） , but significantly inhibited NIH/3T3 cells proliferation induced by TNF-α （ P 〈 0.01 ）. （ 2 ） Compared with the control group, the expression of syndecan-4 protein in NIH/3T3 cells was significantly enhanced by TNF-α at the concentration of 20 ng/mL（P 〈 0.05 ）. IL-10 alone had no effect on the expression of syndecan-4 protein （ P 〉 0.05 ）, but significantly inhibited the expression of syndecan-4 induced by TNF-α （ P 〈 0. 001 ）. Conclusion IL-10 could inhibit expression of syndean-4 protein in NIH/3T3 cells induced by TNF-α in vitro.