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[目的]了解燃煤污染型氟中毒人群外周血中DNA甲基化转移酶1(DNA methylation transferase 1,DNMT1)m RNA转录及蛋白表达情况,探讨其在氟中毒致病机制中的作用。[方法]在知情同意原则下,采集贵州省燃煤污染型氟中毒病区某县的调查对象尿液样本,采用氟离子电极法测定尿氟含量并依据尿氟含量将调查对象(380例)分为正常尿氟(尿氟<1.96 mg/g Cr)组(140例)、低氟(1.96 mg/g Cr≤尿氟<3.92 mg/g Cr)组(93例)、中氟(3.92 mg/g Cr≤尿氟<7.84 mg/g Cr)组(82例)、高氟(尿氟≥7.84 mg/g Cr组(65例);采用实时荧光定量PCR(FQ-PCR)法检测外周血中DNMT1 m RNA转录;采用酶联免疫吸附(ELISA)法检测外周血中DNMT1蛋白表达。[结果]调查对象外周血中DNMT1 m RNA转录,各组间差异具有统计学意义(F=3.125,P<0.05);进一步两两比较分析发现,低、中氟组DNMT1 m RNA转录高于正常尿氟组(P<0.05),且低氟组DNMT1 m RNA转录高于中、高氟组(P均<0.05)。调查对象外周血中DNMT1蛋白表达,各组间差异有统计学意义(F=3.113,P<0.05);进一步两两比较分析发现,低氟组DNMT1蛋白水平高于正常尿氟组(P<0.05),且低氟组DNMT1蛋白水平高于高氟组(P<0.05)。[结论]DNMT1参与氟中毒的发生发展过程,其转录及蛋白表达增强可能是氟骨症发生的早期分子事件。
[Objective] To understand the transcription and protein expression of DNA methylation transferase 1 (DNMT1) m RNA in peripheral blood of people with fluorosis who were poisoned by coal, and to explore its role in the pathogenesis of fluorosis. [Methods] According to the principle of informed consent, urine samples from a county of coal-burning fluorosis in Guizhou Province were collected. Urinary fluoride content was measured by fluoride ion electrode method. According to the urine fluoride content, the surveyed subjects (380 cases) The patients were divided into three groups: normal fluoride (1.96 mg / g Cr urinary fluoride <1.96 mg / g Cr) group (140 cases) / g Cr≤ urine fluoride <7.84 mg / g Cr) group (n = 82) and high fluoride (urine fluoride ≥7.84 mg / g Cr group (n = 65)). The peripheral blood was detected by FQ-PCR DNMT1 m RNA was detected by enzyme-linked immunosorbent assay (ELISA). [Results] DNMT1 m RNA transcripts in the peripheral blood of the investigated subjects were statistically significant (F = 3.125, P (P <0.05). Further analysis showed that DNMT1 m RNA transcripts in low and middle fluoride group were higher than those in normal fluoride group (P <0.05), and DNMT1 m RNA transcription in low fluoride group was higher than that in middle and high fluoride group <0.05) .There was a significant difference between the two groups (F = 3.113, P <0.05), and the DNMT1 protein level in the peripheral blood of the low fluoride group was higher than that of the normal urine fluoride group (P <0.05), and low fluoride (P <0.05). [Conclusion] DNMT1 is involved in the development of fluorosis, and its transcription and protein expression may be the early molecular event of skeletal fluorosis.