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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 To compare the expression of matrix metalloproteinase-3 (MMP-3) and matrix metalloproteinase-3 (TIMP-3) mRNA in blood cells of patients with arsenism with coal-fired arsenism and analyze the relationship between MMP-3 and TIMP- The expression of coal arsenic poisoning patients and its significance, to clarify the coal-based arsenic poisoning a series of clinical symptoms of molecular mechanism. Methods Fifty patients with normal arsenic poisoning and coal samples were collected respectively. Total RNA was extracted by TRIzol-phenol-chloroform method. RNA was determined by UV spectrophotometry. The wavelength (nm) 260/280 (A) was used to identify RNA purity. Real-time quantitative PCR (RT-PCR) was used to determine the mRNA expression of MMP-3 and TIMP-3 in coal-fired arsenic poisoning. The β-actin gene was used as a control. Results The expression of MMP-3mRNA in blood cells of patients with coal-burning arsenism was decreased to 60% of the control group, and the expression of TIMP-3 mRNA was increased to 140% of the control group. The difference was statistically significant (t = 4.39, P <0.01; t = 2.46, P <0.05). Conclusion The MMP-3 and TIMP-3 genes play an important regulatory role in coal-based arsenic poisoning. The decrease of MMP-3mRNA and the increase of TIMP-3mRNA expression may be one of the pathogenesis of coal-burning arsenism.