Ren Pei, Kang Pan, Li Zhaoyang, Zhang Huifang, Niu Qiao
School of Public Health, Shanxi Medical University, Taiyuan 030001, China.
Wei Sheng Yan Jiu. 2017 Jan;46(1):15-20.
To study the impact of the chronic aluminum exposure on Nmethyl-D-aspartate receptor 1( NMDAR1) in the cortex and peripheral blood, and to explore the possibility that whether NMDAR1 of peripheral blood lymphocytes could be taken as a biomarker of aluminum exposure.
Thirty-two cleaning degree, healthy and male SD rats were randomly divided into four groups by weight, i. e. controlgroup, low-dose group, mid-dose group, and high-dose group separately. Different doses of Al Cl3( 20, 120 and 720 mg /kg) were added into the rats' drinking water, and control group was given tap water, each rat approximately drink 10 m L /100 g, the experiment lasted 360 days. Then, plasma aluminum and cortex aluminum were measured by atom absorption spectrometry with graphite furnace( GFAAS), relative expression of NMDAR1 gene was assayed by real-time fluorescence quantitative polymerase chain reaction( RTPCR) and NMDAR1 protein both in the cortex and peripheral blood lymphocytes were assayed by enzyme-linked immunosorbent( ELISA).
Plasma aluminum increased with the increase of Al exposure dose( P < 0. 05), the result of plasma aluminum were69. 88, 83. 10, 87. 06 and 134. 60 μg / L, respectively, plasma aluminum in low-dose, mid-dose and high-dose group were significant increased, compared to the control group. Cortex aluminum increased with the increasing of aluminum dose( P < 0. 05), the result of cortex aluminum were 0. 065, 0. 102, 0. 139 and 0. 228 μg / mg, respectively. The differences among groups were significant( P < 0. 05). The expression of NMDAR1 gene both in the cortex and peripheral blood lymphocytes were reduced with the increasing of aluminum dose( P < 0. 05). The differences of gene expression in the cortex among the three groups were significant statistically( P < 0. 05). The differences of gene expression in peripheral blood lymphocytes among the three groups were significant statistically( P <0. 05), but there was no significantly statistical difference between mid- and high-dose group( P = 0. 167). The protein level of NMDAR1 in the cortex was obviously reduced in high-dose group, compared to that in the control and low-dose group( P < 0. 05). The protein level of NMDAR1 in the peripheral blood lymphocytes were significantly reduced, compared to that in the control and low-dose group( P < 0. 05), the protein level of NMDAR1 in the peripheral blood lymphocytes in the mid- and high-dose group was not different from one another( P = 0. 159).
Chronic aluminum exposure could severely impaire the gene expression and protein expression of NMDAR1 both in the cortex and peripheral blood lymphocytes. With the increasing of the aluminum dose, gene expression and protein expression of NMDAR1 are decreased. The NMDAR1 could be taken as a peripheral biomarker of aluminum exposure for further research.
研究慢性铝暴露对大鼠皮质及外周血中N-甲基-D-天冬氨酸受体1（NMDAR1）的影响，探讨外周血淋巴细胞NMDAR1能否作为铝暴露生物标志物的可能性。
将32只清洁级健康雄性SD大鼠按体重随机分为4组，即对照组、低剂量组、中剂量组和高剂量组。分别在大鼠饮水中加入不同剂量的AlCl₃（20、120和720mg/kg），对照组给予自来水，每只大鼠约饮10mL/100g，实验持续360天。然后，采用石墨炉原子吸收光谱法（GFAAS）测定血浆铝和皮质铝，采用实时荧光定量聚合酶链反应（RT-PCR）检测NMDAR1基因相对表达量，采用酶联免疫吸附法（ELISA）检测皮质及外周血淋巴细胞中NMDAR1蛋白。
血浆铝随铝暴露剂量增加而升高（P<0.05），低剂量组、中剂量组和高剂量组血浆铝分别为69.88、83.10、87.06和134.60μg/L，与对照组相比，低、中、高剂量组血浆铝均显著升高。皮质铝随铝剂量增加而升高（P<0.05），皮质铝分别为0.065、0.102、0.139和0.228μg/mg，组间差异有统计学意义（P<0.05）。皮质及外周血淋巴细胞中NMDAR1基因表达均随铝剂量增加而降低（P<0.05）。皮质中三组基因表达差异有统计学意义（P<0.05）。外周血淋巴细胞中三组基因表达差异有统计学意义（P<0.05），但中剂量组与高剂量组间差异无统计学意义（P=0.167）。高剂量组皮质中NMDAR1蛋白水平与对照组和低剂量组相比明显降低（P<0.05）。外周血淋巴细胞中NMDAR1蛋白水平与对照组和低剂量组相比显著降低（P<0.05），中剂量组与高剂量组外周血淋巴细胞中NMDAR1蛋白水平差异无统计学意义（P=0.159）。
慢性铝暴露可严重损害大鼠皮质及外周血淋巴细胞中NMDAR1的基因表达和蛋白表达。随着铝剂量增加，NMDAR1的基因表达和蛋白表达降低。NMDAR1可作为铝暴露的外周生物标志物进一步研究。
