摘要
目的本研究以燃煤污染型氟人群和原代人成骨细胞为研究对象,检测氟暴露对组蛋白去乙酰化酶1(histone deacetylase1,HDAC1)m RNA转录及蛋白的表达的影响,从人群和细胞两个体系探讨HDAC1基因在地方性氟中毒发生发展中的作用。方法选择贵州省燃煤污染型地方性氟中毒病区295例氟暴露人群及非氟暴露区85例村民作为调查对象,在知情同意的原则下,采集调查对象的尿液及外周血。依据尿氟(UF)含量将调查对象分为正常尿氟组(UF<1.96 mg/g Cr,140例)、低氟组(1.96 mg/g Cr≤UF<3.92 mg/g Cr,93例)、中氟组(3.92 mg/g Cr≤UF<7.84 mg/g Cr,82例)和高氟组(UF≥7.84 mg/g Cr,65例)。同时以0、125、250、500、1 000μmol/L氟化钠处理人原代成骨细胞72 h。采用实时荧光定量PCR(q RT-PCR)检测氟暴露人群外周血及染氟人原代成骨细胞HDAC1 m RNA水平,以酶联免疫吸附试验(ELISA)和免疫印迹法(Western-blot)分别检测外周血、成骨细胞HDAC1蛋白表达。结果调查对象外周血HDAC1 m RNA转录水平各组间比较差异有统计学意义(F=16.502,P<0.05),低、中、高尿氟组HDAC1 m RNA水平均低于正常尿氟组(P<0.05);外周血HDAC1蛋白表达,各组间比较差异有统计学意义(F=4.885,P<0.05),低、中、高尿氟组HDAC1蛋白含量均高于正常尿氟组(P<0.05)。氟化钠处理人成骨细胞72 h后,HDAC1 m RNA水平,组间比较差异有统计学意义(F=226.81,P<0.05),随着染氟浓度的升高,HDAC1 m RNA转录水平逐渐升高(P<0.05);HDAC1蛋白表达组间比较差异有统计学意义(F=20.60,P<0.05);随着染氟浓度的升高,HDAC1蛋白表达水平逐渐升高(P<0.05)。结论 HDAC1参与氟中毒的发生发展过程,其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 histone deacetylase 1（HDAC1） gene m RNA transcription and protein expression influenced by fluoride exposure from coal burning in population and human primary osteoblasts cultured in vitro,and explore the role of HDAC1 in fluorosis pathogenesis. Methods A total of 295 cases were chosen as the fluoride exposure group in a coal-burning endemic fluorosis area in Guizhou province,and 85 controls were selected in non-fluoride contaminated area.Following the principle of informed consent, the urine and blood samples were collected. According to the content of urine fluoride（UF）, the investigated individuals were stratified into normal（UF ≤1.96 mg/g Cr, 140 cases）, low（1.96 mg/g Cr ≤UF3.92 mg/g Cr, 93 cases）, medium（3.92 mg/g Cr≤UF7.84 mg/g Cr, 82 cases）, and high UF groups（UF≥7.84 mg/g Cr, 65 cases）. The primary osteoblasts were treated with sodium fluoride at 0, 125, 250, 500 and 1 000 μmol/L respectively for 72 h.The m RNA transcription of HDAC1 in respondents＇ peripheral blood and osteoblasts were detected by quantitative real-time PCR（q RT-PCR）. The HDAC1 protein in peripheral blood and osteoblasts were detected by enzyme linked immune sorbent assay（ELISA） and Western blot, respectively. Results The transcription levels of HDAC1 m RNA in peripheral blood samples significantly varied among the groups（F=16.502, P〈0.05）. The results of pair-wise comparison found that the low, middle and high UF groups presented higher expression of m RNA transcription than the normal UF group（P〈0.05）; The contents of HDAC1 protein in peripheral blood samples were different among the groups（F =4.885, P〈0.05）. The results of pair-wise comparison showed that the normal UF group presented lower HDAC1 protein content than the low, middle and high UF groups（F =16.502, P〈0.05）.The transcription levels of HDAC1 m RNA in osteoblasts treated with Na F for 72 h significantly varied among the groups（F=226.81,P〈0.05）. The results of pair-wise comparison found that with the increase of concentrations of Na F, the expression of HDAC1 m RNA were correspondingly up-regulated in a dose-dependent manner. The expression of HDAC1 protein in osteoblasts significantly varied among the groups（F =20.60,P〈0.05）. The results of pair-wise comparison found that with the increase of Na F exposure, the expression of HDAC1 protein were correspondingly up-regulated in a dosedependent manner. Conclusion The increased HDAC1 protein expression is involved in the fluorosis pathogenesis and may be the early molecular events of fluorosis.
作者
明娟
严威敏
廖玉丹
汪希兰
潘雪莉
MING Juan;YAN Wei-min;LIAO Yu-dan;WANG Xi-lan;PAN Xue-li(School of Public Health, Guizhou Medical University, Gttiyang , Guizhott 550000, Chin)
出处
《环境与健康杂志》
CAS
北大核心
2018年第2期104-107,共4页
Journal of Environment and Health
基金
国家自然科学基金(81260418)
贵州省优秀科技教育人才省长基金(黔省专合字[2011]54号)
贵阳市科技计划项目(筑科合同[2017]30-6号)
