發(fā)布時(shí)間：2019-02-18 07:51

【摘要】：[目的]探討鎘對(duì)人胚胎神經(jīng)干細(xì)胞增殖能力的影響和對(duì)金屬硫蛋白(MT)各亞型的誘導(dǎo)表達(dá)。[方法]以0、1.25、2.5、5μmol/L氯化鎘(Cd Cl2)染毒人胚胎神經(jīng)干細(xì)胞24 h后,以四甲基偶氮唑鹽(MTT)法測(cè)定細(xì)胞活力,5-乙炔基-2′脫氧尿苷(Ed U)法測(cè)定細(xì)胞增殖能力,分光光度法測(cè)定培養(yǎng)上清中乳酸脫氫酶活性(LDH),應(yīng)用逆轉(zhuǎn)錄多聚酶鏈反應(yīng)(RT-PCR)檢測(cè)鎘對(duì)人胚胎神經(jīng)干細(xì)胞MT 10個(gè)亞型的m RNA表達(dá)的影響。[結(jié)果]與對(duì)照組相比,濃度為2.5、5μmol/L Cd Cl2染毒組細(xì)胞活力明顯降低,差異具有統(tǒng)計(jì)學(xué)意義(P0.01);且細(xì)胞相對(duì)存活率與Cd Cl2濃度呈負(fù)相關(guān)(r=-0.98,P0.01)。1.25μmol/L及以上鎘可明顯抑制DNA的復(fù)制(P0.05),且細(xì)胞增殖與Cd Cl2濃度對(duì)數(shù)值呈負(fù)相關(guān)(r=-0.91,P0.01)。與對(duì)照組相比,各染毒組細(xì)胞培養(yǎng)上清中LDH水平均高于對(duì)照組,差異有統(tǒng)計(jì)學(xué)意義(P0.01),且細(xì)胞培養(yǎng)上清中LDH水平與Cd Cl2濃度呈正相關(guān)(r=0.97,P0.01)。與對(duì)照組相比,MT基因各亞型的m RNA表達(dá)水平隨著Cd Cl2染毒濃度的升高而升高,差異均有統(tǒng)計(jì)學(xué)意義(均P0.05)。[結(jié)論]鎘可引起人神經(jīng)胚胎干細(xì)胞自我更新功能的抑制,表現(xiàn)為細(xì)胞活力和增殖能力的下降,細(xì)胞培養(yǎng)液中LDH含量的上升,細(xì)胞MT基因各亞型m RNA表達(dá)水平的明顯升高。
[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.
【作者單位】： 復(fù)旦大學(xué)公共衛(wèi)生學(xué)院公共衛(wèi)生安全國(guó)家重點(diǎn)實(shí)驗(yàn)室;
【基金】：國(guó)家自然科學(xué)基金項(xiàng)目(編號(hào):81472996) 上海自然科學(xué)基金項(xiàng)目(編號(hào):10ZR1401700)
【分類號(hào)】：R114

【參考文獻(xiàn)】

相關(guān)期刊論文 前4條

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