镉对人胚胎神经干细胞增殖抑制及对金属硫蛋白各亚型的诱导表达
发布时间：2019-02-18 07:51
【摘要】：[目的]探讨镉对人胚胎神经干细胞增殖能力的影响和对金属硫蛋白(MT)各亚型的诱导表达。[方法]以0、1.25、2.5、5μmol/L氯化镉(Cd Cl2)染毒人胚胎神经干细胞24 h后,以四甲基偶氮唑盐(MTT)法测定细胞活力,5-乙炔基-2′脱氧尿苷(Ed U)法测定细胞增殖能力,分光光度法测定培养上清中乳酸脱氢酶活性(LDH),应用逆转录多聚酶链反应(RT-PCR)检测镉对人胚胎神经干细胞MT 10个亚型的m RNA表达的影响。[结果]与对照组相比,浓度为2.5、5μmol/L Cd Cl2染毒组细胞活力明显降低,差异具有统计学意义(P0.01);且细胞相对存活率与Cd Cl2浓度呈负相关(r=-0.98,P0.01)。1.25μmol/L及以上镉可明显抑制DNA的复制(P0.05),且细胞增殖与Cd Cl2浓度对数值呈负相关(r=-0.91,P0.01)。与对照组相比,各染毒组细胞培养上清中LDH水平均高于对照组,差异有统计学意义(P0.01),且细胞培养上清中LDH水平与Cd Cl2浓度呈正相关(r=0.97,P0.01)。与对照组相比,MT基因各亚型的m RNA表达水平随着Cd Cl2染毒浓度的升高而升高,差异均有统计学意义(均P0.05)。[结论]镉可引起人神经胚胎干细胞自我更新功能的抑制,表现为细胞活力和增殖能力的下降,细胞培养液中LDH含量的上升,细胞MT基因各亚型m RNA表达水平的明显升高。
[Abstract]:[objective] to investigate the effect of cadmium on the proliferation of human embryonic neural stem cells and the expression of metallothionein (MT) subtypes. [methods] Human embryonic neural stem cells were exposed to 2.5 渭 mol/L cadmium chloride (Cd Cl2) for 24 h, and the cell viability was determined by tetramethyl azolium salt (MTT) assay. Determination of cell proliferation by 5-acetyl-2 'deoxyuridine (Ed U) assay and determination of lactate dehydrogenase activity (LDH), in culture supernatant by spectrophotometry Reverse transcriptase polymerase chain reaction (RT-PCR) was used to detect the effect of cadmium on the expression of m RNA in 10 MT subtypes of human embryonic neural stem cells. [results] compared with the control group, the cell viability in the 2.5 渭 mol/L Cd Cl2 group was significantly lower than that in the control group (P0.01). The relative survival rate of cells was negatively correlated with the concentration of Cd Cl2 (r-0.98U, P0.01). Cadmium (1.25 渭 mol/L or above) significantly inhibited the replication of DNA (P0.05), and the cell proliferation was negatively correlated with the concentration of Cd Cl2 (r-0.91). P0.01) Compared with the control group, the level of LDH in the supernatant of cell culture in each exposed group was higher than that in the control group (P0.01), and the level of LDH in the supernatant of cell culture was positively correlated with the concentration of Cd Cl2 (r 0.97% P0.01). Compared with the control group, the expression of m RNA in each subtype of MT gene increased with the increase of Cd Cl2 concentration, and the difference was statistically significant (P0.05). [conclusion] cadmium can inhibit the self-renewal function of human neural embryonic stem cells, such as the decrease of cell viability and proliferation ability, the increase of LDH content in cell culture medium, and the increase of m RNA expression of MT gene subtypes.
【作者单位】： 复旦大学公共卫生学院公共卫生安全国家重点实验室;
【基金】：国家自然科学基金项目(编号:81472996) 上海自然科学基金项目(编号:10ZR1401700)
【分类号】：R114
[Abstract]:[objective] to investigate the effect of cadmium on the proliferation of human embryonic neural stem cells and the expression of metallothionein (MT) subtypes. [methods] Human embryonic neural stem cells were exposed to 2.5 渭 mol/L cadmium chloride (Cd Cl2) for 24 h, and the cell viability was determined by tetramethyl azolium salt (MTT) assay. Determination of cell proliferation by 5-acetyl-2 'deoxyuridine (Ed U) assay and determination of lactate dehydrogenase activity (LDH), in culture supernatant by spectrophotometry Reverse transcriptase polymerase chain reaction (RT-PCR) was used to detect the effect of cadmium on the expression of m RNA in 10 MT subtypes of human embryonic neural stem cells. [results] compared with the control group, the cell viability in the 2.5 渭 mol/L Cd Cl2 group was significantly lower than that in the control group (P0.01). The relative survival rate of cells was negatively correlated with the concentration of Cd Cl2 (r-0.98U, P0.01). Cadmium (1.25 渭 mol/L or above) significantly inhibited the replication of DNA (P0.05), and the cell proliferation was negatively correlated with the concentration of Cd Cl2 (r-0.91). P0.01) Compared with the control group, the level of LDH in the supernatant of cell culture in each exposed group was higher than that in the control group (P0.01), and the level of LDH in the supernatant of cell culture was positively correlated with the concentration of Cd Cl2 (r 0.97% P0.01). Compared with the control group, the expression of m RNA in each subtype of MT gene increased with the increase of Cd Cl2 concentration, and the difference was statistically significant (P0.05). [conclusion] cadmium can inhibit the self-renewal function of human neural embryonic stem cells, such as the decrease of cell viability and proliferation ability, the increase of LDH content in cell culture medium, and the increase of m RNA expression of MT gene subtypes.
【作者单位】： 复旦大学公共卫生学院公共卫生安全国家重点实验室;
【基金】：国家自然科学基金项目(编号:81472996) 上海自然科学基金项目(编号:10ZR1401700)
【分类号】：R114
【参考文献】
相关期刊论文 前4条
1 安红敏;郑伟;高扬;;镉的健康危害及干预治疗研究进展[J];环境与健康杂志;2007年09期
2 常秀丽;金泰^，
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