赵越, 王林平, 牛侨. 麦芽酚铝暴露对大鼠脑皮质神经元微小RNA-29a和BACE1表达的影响[J]. 环境与职业医学, 2016, 33(5): 438-443. DOI: 10.13213/j.cnki.jeom.2016.15604
引用本文: 赵越, 王林平, 牛侨. 麦芽酚铝暴露对大鼠脑皮质神经元微小RNA-29a和BACE1表达的影响[J]. 环境与职业医学, 2016, 33(5): 438-443. DOI: 10.13213/j.cnki.jeom.2016.15604
ZHAO Yue, WANG Lin-ping, NIU Qiao. Influence on MicroRNA-29a and BACE1 Expression in Rat Cortical Neurons Exposed to Aluminum Maltolate[J]. Journal of Environmental and Occupational Medicine, 2016, 33(5): 438-443. DOI: 10.13213/j.cnki.jeom.2016.15604
Citation: ZHAO Yue, WANG Lin-ping, NIU Qiao. Influence on MicroRNA-29a and BACE1 Expression in Rat Cortical Neurons Exposed to Aluminum Maltolate[J]. Journal of Environmental and Occupational Medicine, 2016, 33(5): 438-443. DOI: 10.13213/j.cnki.jeom.2016.15604

麦芽酚铝暴露对大鼠脑皮质神经元微小RNA-29a和BACE1表达的影响

Influence on MicroRNA-29a and BACE1 Expression in Rat Cortical Neurons Exposed to Aluminum Maltolate

  • 摘要: 目的

    探讨麦芽酚铝暴露对大鼠脑皮质神经元微小RNA(miR)-29a和BACE1表达的影响。

    方法

    取新生24 h内的SD大鼠乳鼠的脑皮质进行原代神经元培养,培养至第3天时加入10 μmol/L的阿糖胞苷,应用免疫组化法检测神经元特异性表达蛋白的表达来进行神经元纯度检测,应用不同剂量麦芽酚铝染毒原代神经元,剂量分别为空白对照组(0 μmol/L)、低剂量组(20 μmol/L)、中剂量组(40 μmol/L)、高剂量组(80 μmol/L)4组,继续培养24 h收集细胞,荧光实时定量PCR检测miR-29aBACE1 mRNA表达水平,ELISA法检测BACE1蛋白含量。

    结果

    免疫组化结果显示:神经元纯度达到95%。荧光实时定量PCR结果显示:对照组、低剂量组、中剂量组、高剂量组BACE1 mRNA的相对表达量分别为1.00±0.00、1.24±0.32、1.64±0.12、1.87±0.06,与对照组相比,中、高剂量组的表达水平升高(P < 0.05);而miR-29a的相对表达量分别为1.00±0.00、0.98±0.17、0.47±0.03、0.34±0.08,与对照组相比,中、高剂量组的表达降低(P < 0.05);ELISA结果显示:对照组、低剂量组、中剂量组、高剂量组BACE1表达量分别为(406.89±57.47)、(419.52±49.38)、(497.10±4.79)、(508.83±44.59)ng/L,与对照组相比,中、高剂量组的表达水平升高(P < 0.05)。

    结论

    铝可能通过抑制miR-29a的表达,从而负向调控BACE1表达的增多,导致β淀粉样蛋白沉积。

     

    Abstract: Objective

    To assess the influence of aluminum maltolateAl(mal)3 exposure on microRNA(miR)-29a and BACE1 expression in rat cortical neurons.

    Methods

    The cerebral cortex of newborn SD rats (≤24 h old) was used for primary neuronal cultures, and 10 μmol/L of cytarabine was added on the third day. Neuron-specific class Ⅲ β-tublin was used to detect neuron purity by immunohistochemical assay. Primary neurons were exposed to different doses of Al(mal)3 and divided into control (0 μmol/L), low dose (20 μmol/L), middle dose (40 μmol/L), and high dose (80 μmol/L) groups. After 24 h Al(mal)3 exposure, the cells were collected for determining miR-29a and BACE1 mRNA expressions by real-time fluorescence quantitative PCR, as well as BACE1 protein expression by ELISA.

    Results

    The results of immunohistochemical assay showed the purity of neurons reaching 95%. The real-time fluorescence quantitative PCR results showed that the relative expressions of BACE1 mRNA in the control, low dose, middle dose, and high dose groups were 1.00±0.00, 1.24±0.32, 1.64±0.12, and 1.87±0.06, respectively; compared with the control group, the expression of BACE1 mRNA were increased in the middle dose group and the high dose group (P < 0.05). However, the relative expressions of miR-29a were 1.00±0.00, 0.98±0.17, 0.47±0.03, and 0.34±0.08, respectively; compared with the control group, the expressions of miR-29a were decreased in the middle dose group and the high dose group (P < 0.05). The ELISA results showed that the expressions of BACE1 in the four groups were (406.89±57.47), (419.52±49.38), (497.10±4.79), and (508.83±44.59) ng/L, respectively; compared with the control group, the levels were increased in the middle and high dose groups (P < 0.05).

    Conclusion

    Aluminum might regulate the expression of BACE1 negatively and result in amyloid β-protein deposition through inhibiting the expression of miR-29a.

     

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