基质金属蛋白酶-9对百草枯中毒患者预后的预测价值

The predictive value of matrix metalloproteinase-9 for prognosis of patients with paraquat poisoning

目的观察口服百草枯（PQ）中毒患者外周血基质金属蛋白酶-9（MMP-9）的基因表达情况，并评价其对PQ中毒预后的预测价值。方法选择2013年1月至2014年6月临沂市人民医院收治的口服PQ中毒患者37例，根据28d存活情况分为存活组（26例）和死亡组（11例）；同期选择10例健康体检者作为对照。PO中毒患者于入院1d、3d取外周静脉血3mL，健康对照者于体检当日取空腹静脉血3mL，采用反转录-聚合酶链反应（RT—PCR）检测外周血单核细胞（PBMCs）中MMP-9的基因表达，采用酶联免疫吸附试验（ELISA）测定血清MMP-9含量；采用高效液相色谱法（HPLC）检测PQ中毒患者血清PQ水平，并记录服毒量。采用Spearman相关法分析服毒量、血清PQ水平与外周MMP-9的相关性。绘制受试者工作特征曲线（ROC），评价外周血MMP-9水平对口服PQ中毒患者28d预后的预测价值。结果37例口服PQ中毒患者人院1d血清P0水平为（2．60±1．29）mg／L，服毒量50．0（7．5，60．0）mL。两组患者PBMCs中MMP-9基因表达和血清MMP-9含量均明显高于健康对照组，且随时间延长逐渐升高，以死亡组升高更为显著[PBMCs中MMP-9基因表达（A值）：1d为2．84±1．16比0．95±0．23，3d为4．22±1．75比1．29±0．30；血清MMP-9含量（μg/L）：1d为2791．48±1230．88比807．81±279．86，3d为4384．21±1781．97比1131．14±291．76，均P〈0．05）。相关性分析显示：口服PQ中毒患者血清PQ水平、服毒量与外周血PBMCs中MMP-9基因表达和血清MMP-9含量均呈显著正相关（均P=0．000）。ROC曲线分析显示：人院1dMMP-9基因表达和3d血清MMP-9含量对口服PQ中毒患者28d预后有预测价值，ROC曲线下面积（AUC）分别为0．820和0．776。当人院1dMMP-9基因表达量截断值为0．90时，其敏感度和特异度分别为80．00％、63．64％；入院3d血清MMP-9含量截断值为904．36μg／L时，其敏感度和特异度分别为80．00％、72．73％。结论口服PQ中毒可导致患者体内MMP-9基因表达和血清MMP-9含量升高，MMP-9基因表达对口服PQ中毒预后有预测价值。

Objective To detect the expression of matrix metalloproteinase-9 （MMP-9） gene in peripheral blood of patients with oral paraquat （PQ） poisoning and evaluate its predictive value for their prognosis. Methods Thirty-seven cases of oral PQ poisoning admitted to Linyi People＇s Hospital from January 2013 to June 2014 were enrolled, and they were divided into survival group （26 cases） and death group （11 cases） according to the survival situation in 28 days after poisoning; a healthy control group included 10 healthy people selected in the same period. The peripheral blood 3 mL was collected from each PQ patient on the 1st and 3rd day after admission, and in the healthy control group, 3 mL peripheral venous blood was obtained under fast on the day for physical examination. The MMP-9 gene expression of peripheral blood mononuclear cells （PBMCs） was detected by reverse transcription-polymerase chain reaction （RT-PCR） methods; the serum MMP-9 concentration was determined by using enzyme-linked immunosorbent assay （ELISA）; the serum PQ level was detected by high performance liquid chromatography （HPLC）, and the amount of poison orally taken was recorded. The correlations between PQ amount orally taken, serum PQ level and MMP-9 expression were analyzed by Spearman correlation analysis. A receiver operating characteristic （ROC） curve was drawn to evaluate the predictive value of peripheral blood MMP-9 level for the 28-day prognosis of PQ poisoning patients. Results After admission the 1 day serum PQ level was （2.60±1.29） mg/L, and the amount of poison taken was 50.0 （7.5, 60.0） mL in the 37 patients with oral PQ poisoning. The MMP-9 gene expression level in PBMCs and serum MMP-9 protein level of both PQ poisoning groups were significantly higher than those of healthy control group, and the levels were gradually increased with the extension of poisoning time; the degrees of elevation in death group were more significant [the PBMCs＇ MMP-9 gene expression （A value）： 2.84 ＋ 1.16 vs. 0.95 ＋ 0.23 on the 1st poisoning day, 4.22±1.75 vs. 1.29±0.30 on the 3rd poisoning day; serum MMP-9 concentration （μg/L）： 2791.48±1230.88 vs. 807.81±279.86 on the 1st poisoning day, 4384.21 ± 1 781.97 vs. 1 131.14±291.76 on the 3rd poisoning day, all P 〈 0.05]. Correlation analysis showed： there were significant positive correlations of oral PQ amount, serum PQ concentration to the MMP-9 gene expression in PBMCs and serum MMP-9 protein concentration in patients with oral PQ poisoning （all P = 0.000）. ROC curve analysis showed： the MMP-9 gene expression in PBMCs on the 1st day and the serum MMP-9 content on the 3rd day after admission had predictive value for 28-day prognosis in patients with oral PQ poisoning, and the ROC areas under the curve （AUC） was 0.820 and 0.776 respectively. When the cutoff value of MMP-9 gene expression level on the 1st day after admission was 0.90, the predictive sensitivity and specificity were 80.00% and 63.64% respectively; when the cutoff value of serum MMP-9 protein content on the 3rd day after admission was 904.36μg/L, the predictive sensitivity and specificity were 80.00% and 72.73% respectively. Conclusion Oral PQ poisoning can lead to the MMP-9 gene expression in PBMCs and elevation of serum MMP-9 protein level in the body, and the MMP-9 gene expression has predictive value for the prognosis of patients with oral PQ poisoning.