Objective: To investigate the expression of tumor necrosis factor alpha-inducible protein 8 like-2 (TIPE2) and tissue factor (TF) in patients with bronchial asthma. And to explore the regulation of TIPE2 on TF. Methods: Sixty-five asthmatic patients and 40 healthy controls were selected from the First Affiliated Hospital of Zhengzhou University from July to November, 2017. The expression of TIPE2 and TF in peripheral blood mononuclear cells of asthmatic patients and healthy controls were detected by Western blot.The level of TF protein in plasma was detected by enzyme-linked immunosorbent assay. The changes of TIPE2 and TF mRNA expression in THP-1 cells stimulated by house dust mite extract were detected by real-time fluorescence quantitative PCR (RT-PCR). The recombinant adenovirus Adv-TIPE2 was constructed and transfected into THP-1 cells and the effect of over-expression TIPE2 on TF expression in THP-1 cells was detected by RT-PCR. Results: The relative level of TIPE2 protein in asthmatic patients and healthy controls was 0.025±0.010 and 0.087±0.070, while that of TF was 0.40±0.27 and 0.15±0.10, respectively. Compared with healthy controls, the levels of TIPE2 protein decreased and TF protein increased in asthmatic patients, the differences were statistically significant (t=-5.06, 9.04, P<0.05) . TIPE2 protein level was negatively correlated with TF protein level (r=-0.460 3, P<0.05). The house dust mite extract reduced the expression level of TIPE2 mRNA in THP-1 cells, but increased the level of TF mRNA expression. When the concentration was 1 μg/ml, the change of TIPE2 mRNA was the most obvious at 4 h (P<0.05). The recombinant adenovirus Adv-TIPE2 was successfully constructed. The level of TF mRNA expression in THP-1 cells over-expressing TIPE2 gene was reduced (P<0.05). Conclusion: TIPE2, a negative regulator of inflammation, has a negative control effect on TF, and may be involved in the hyper-coagulable state of bronchial asthma by regulating TF expression.
目的： 研究支气管哮喘患者体内肿瘤坏死因子－α诱导蛋白8样分子－2(TIPE2)和组织因子(TF)表达情况，初步探讨TIPE2对TF的调控作用。 方法： 选取2017年7至11月郑州大学第一附属医院支气管哮喘患者65例，健康对照者40名。免疫印迹法检测哮喘患者和健康对照者外周血单个核细胞中TIPE2、TF的表达情况，酶联免疫法检测血浆中TF蛋白表达水平。实时荧光定量PCR(RT－PCR)检测屋尘螨提取液刺激THP－1细胞中TIPE2、TF mRNA表达水平的变化。构建重组腺病毒Adv－TIPE2，转染THP－1细胞，RT－PCR检测TIPE2过表达对THP－1细胞中TF表达的影响。 结果： 哮喘患者组与健康对照组TIPE2蛋白相对表达水平分别为0.025±0.010、0.087±0.070，TF蛋白相对表达水平分别为0.40±0.27、0.15±0.10，与健康对照者相比，哮喘患者体内TIPE2蛋白水平降低，TF蛋白水平升高，差异均有统计学意义(t＝－5.06、9.04，均P<0.05)，且TIPE2蛋白水平与TF蛋白水平呈负相关(r＝－0.460 3, P<0.05)。屋尘螨提取液可降低THP－1细胞中TIPE2 mRNA的表达水平，而升高TF mRNA的表达水平，当浓度为1 μg/ml，刺激4 h时变化最明显(P<0.05)。成功构建了重组腺病毒Adv－TIPE2，过表达TIPE2基因的THP－1细胞内TF mRNA表达水平降低(P<0.05)。 结论： 炎症负性调节分子TIPE2对TF具有负调控作用，可能通过调节TF的表达参与支气管哮喘的高凝状态。.
Keywords: Bronchial asthma; Tissue factor; Tumor necrosis factor-2 induced protein 8 like-2.