MMP-3和TIMP-3在燃煤型砷中毒患者血细胞中的表达

Expression of MMP-3 and TIMP-3 gene in the patients suffered from coal-burning arsenism

目的比较燃煤型砷中毒患者与正常人血细胞中基质金属蛋白酶-3(MMP-3)和基质金属蛋白酶抑制因子-3(TIMP-3)mRNA表达,分析MMP-3、TIMP-3基因在燃煤型砷中毒患者中的表达及其意义,阐明燃煤型砷中毒出现一系列临床症状的分子机制。方法分别采集燃煤型砷中毒患者与正常人外周血标本各50例,采用TRIzol-酚-氯仿一步法提取总RNA。紫外分光光度法测定RNA,波长(nm)260/280(A)鉴定RNA纯度。实时荧光定量PCR(RT-PCR)技术测定燃煤型砷中毒MMP-3、TIMP-3基因mRNA的表达,以β-肌动蛋白(β-actin)基因作为质控。结果燃煤型砷中毒患者血细胞中MMP-3mRNA表达降低,为对照组的60%;TIMP-3mRNA表达升高,为对照组的140%,二者比较差异均有统计学意义(t=4.39,P<0.01;t=2.46,P<0.05)。结论MMP-3、TIMP-3基因在燃煤型砷中毒代谢中发挥了重要的调控作用。MMP-3mRNA表达降低和TIMP-3mRNA表达升高,可能是燃煤型砷中毒发病机制之一。

Objective In order to elucidate the molecular mechanism of arsenism caused by burning coal by comparing gene expression of matrix metallonrotease-3(MMP-3) and its tissue inhibitors of metallonrotease-3(TIMP-3)between patients with arsenism and normal people. Methods RNA was extracted by the TRIzol-Phenol-Chloroform method from leukocytes of 50 patients suffered from coal-burning arsenism and 50 healthy volunteers. The quantity of the RNA was determined by spectrophotometry and its purity was judged at a ratio of 260/280(A). Then real-time PCR(RT-PCR) was used to measure the mRNA expression of MT in the coal-burning arsenism, β-actin used as a quality control. Results The mRNA expression of MMP-3 reduced in the patients approximately by 40% comparing with the normal controls with a statistical difference(t = 4.39, P < 0.01). But the mRNA expression of TIMP-3 increased in the patients approximately by 140% comparing with the normal controls with a statistical difference(t = 2.46, P < 0.05). Conclusions MMP-3 and TIMP-3 play a regulating role in the metabolism of coal-burning arsenism. The decrease of the mRNA expression of MT in the patients with coal-burning arsenism may be related to the arsenism mechanism.