摘要
目的探讨三磷酸肌醇受体Ⅲ型(IP3RⅢ)DNA甲基化对硫酸铍(BeSO4)致人胚肺成纤维细胞(MRC-5细胞)损伤的影响。方法建立离体细胞培养实验模型,以终浓度分别为1.0、10.0、100.0μmol/L的BeSO4染毒MRC-5细胞24 h(设为低、中、高剂量染毒组),对照组不使用BeSO4染毒。采用巢式降落式甲基化特异性聚合酶链式反应(PCR)法检测IP3RⅢDNA甲基化的变化,实时定量PCR检测IP3RⅢmRNA的表达,激光共聚焦显微镜检测细胞钙离子浓度([Ca2+]i)。结果高剂量组IP3RⅢDNA甲基化值分别低于对照组、低剂量组和中剂量组[(0.26±0.07)vs(0.48±0.02)、(0.44±0.04)、(0.37±0.03),P<0.01,P<0.01,P<0.05];低、中、高剂量组IP3RⅢmRNA表达量均高于对照组[(0.35±0.08)、(0.31±0.05)、(0.71±0.09)vs(0.03±0.01),P<0.01],高剂量组IP3RⅢmRNA表达量分别高于低、中剂量组[(0.71±0.09)vs(0.35±0.08)、(0.31±0.05),P<0.01];高剂量组细胞内[Ca2+]i平均荧光强度高于对照组[(6 171.66±601.72)vs(3 618.33±559.26),P<0.05]。结论 IP3RⅢDNA低甲基化致细胞内IP3RⅢmRNA表达增加,[Ca2+]i增加,可能是BeSO4致MRC-5细胞损伤的重要机制之一。
Objective To explore the effects of inositol trisphosphate receptor Ⅲ （ IP3 R Ⅲ ） methylation in the injury of human embryonic hmg fibrobiast （ MRC-5 cell） induced by beryllium sulfate （ BeSOg ）. Methods Experimental model of cell culture in vitro was established, MRC-5 cells were exposed to final concentrations of 0. 0, 1.0, 10. 0, 100. 0 μmol/L of BeSO4 for 24 hours （control-, low-, medium- and high- dose groups）. DNA methylation status of IP3RⅢ was detected by nMS-polymerase chain reaction （PCR） , the expression of IP3RⅢ mRNA was detected by real-time PCR, intracellular cytoplasmic free Ca2＋ concentration （[ Ca2＋]i） was detected by laser scanning confocal microscope. Results IP3RⅢ DNA methylation values in high dose group were lower than those in the control, low dose and medium dose groups [ （0. 26 ±0.07） vs （0.48±0.02）, （0. 44±0.04）, （0.37±0.03）, P〈0.01, P〈0.01, P〈0.05]. IP3R~[ mRNA expression in low, medium and high dose groups were higher than those of the control [ （ 0. 35 ±0.08 ） , （ 0. 31 ±0. 05 ） , （ 0. 71 ±0. 09） vs （0. 03 ±0. 01 ）, P 〈0. 01 ]. IP3RⅢ mRNA expression in high dose group was higher than those in low and medium dose groups [ （0. 71 ±0. 09） vs （0. 35 ±0. 08 ）, （0. 31 ±0. 05 ）, P 〈 0. 01 1. [ Ca2 ＋ ] i mean fluorescence intensity in high dose group was higher than that of the control group [ （6 171.66 ±601.72） vs （3 618.33 ±559. 26）, P 〈 0. 05 ]. Conclusion The hypomethylation of IP3R Ⅲ DNA methylation which results in the increase of the intracellular IP3R ⅢmRNA expression and [ Ca2＋] i concentrations by BeSO4 may be one important mechanism of MRC-5 injury.
出处
《中国职业医学》
CAS
北大核心
2013年第5期398-402,共5页
China Occupational Medicine
基金
宁夏医科大学"博士学位建设学科"开放课题(KF2010-01)
宁夏高等学校科学技术研究重点项目(NGY2012048)
宁夏回族自治区卫生厅科研项目(2012011)
关键词
三磷酸肌醇受体
DNA甲基化
硫酸铍
人胚肺成纤维细胞
Inositol trisphosphate receptor
DNA methylation
Beryllium sulfate
Human embryonic lung fibroblast
