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地塞米松对培养的胎鼠大脑皮质神经元细胞质中动力蛋白重链和动力蛋白激活蛋白表达的影响

[Effect of dexamethasone on the expression of Dynein heavy chain and Dynactin in the cytoplasm of fetal rat cerebral cortical neurons cultured ].

作者信息

Cheng Lin, Xie Zi-Yun, Li Jian, Bo Tao

机构信息

Department of Pediatrics, Third Xiangya Hosptial, Central South University, Changsha 410011, China.

出版信息

Zhongguo Dang Dai Er Ke Za Zhi. 2021 Jun;23(6):639-644. doi: 10.7499/j.issn.1008-8830.2103151.

Abstract

OBJECTIVE

To study the effect of dexamethasone (DEX) on the expression of Dynein heavy chain (DHC) and Dynactin in the cytoplasm of fetal rat cerebral cortical neurons cultured .

METHODS

Primary cerebral cortical neurons of fetal rats were cultured and were used to establish a cellular model of DEX intervention. According to the final concentration of DEX, the neurons were divided into three groups:control (without DEX), 0.1 μmol/L DEX, and 1.0 μmol/L DEX. On days 1, 3, and 7 after intervention, the quantitative PCR was used to observe the effect of DEX on the mRNA expression of DHC and Dynactin. The Western blot was used to observe the effect of DEX on the protein expression of DHC and Dynactin.

RESULTS

There was no significant difference in the mRNA expression levels of DHC and Dynactin among the three groups at all time points ( > 0.05). On day 7 after DEX intervention, the protein expression of DHC in the 1.0 μmol/L DEX group gradually increased and reached the peak over time, which was significantly higher than that in the control and 0.1 μmol/L DEX groups ( < 0.05). The control and 0.1 μmol/L DEX groups had a significant increase in the protein expression of Dynactin from day 1 to days 3 and 7 after DEX intervention ( < 0.05). The control group had a significant increase in the protein expression of Dynactin from day 3 to day 7 after intervention ( < 0.05), while the 0.1 μmol/L DEX group had a significant reduction in the protein expression of Dynactin from day 3 to day 7 after intervention ( < 0.05). On days 3 and 7 after DEX intervention, the 0.1 μmol/L DEX and 1.0 μmol/L DEX groups had a significantly lower protein expression level of Dynactin in the cerebral cortical neurons than the control group ( < 0.05). On day 7 after DEX intervention, the 1.0 μmol/L DEX group had a significantly lower protein expression level of Dynactin than the 0.1 μmol/L DEX group ( < 0.05).

CONCLUSIONS

DEX affects the protein expression of DHC and Dynactin in the fetal rat cerebral cortical neurons cultured , possibly in a concentration- and time-dependent manner.

摘要

目的

研究地塞米松(DEX)对体外培养的胎鼠大脑皮质神经元胞质中动力蛋白重链(DHC)和动力蛋白激活蛋白表达的影响。

方法

培养胎鼠原代大脑皮质神经元,建立DEX干预细胞模型。根据DEX终浓度将神经元分为三组:对照组(无DEX)、0.1 μmol/L DEX组和1.0 μmol/L DEX组。干预后第1、3、7天,采用定量PCR观察DEX对DHC和动力蛋白激活蛋白mRNA表达的影响;采用蛋白质印迹法观察DEX对DHC和动力蛋白激活蛋白蛋白表达的影响。

结果

各时间点三组间DHC和动力蛋白激活蛋白的mRNA表达水平差异均无统计学意义(P>0.05)。DEX干预后第7天,1.0 μmol/L DEX组DHC蛋白表达随时间逐渐升高并达到峰值,显著高于对照组和0.1 μmol/L DEX组(P<0.05)。对照组和0.1 μmol/L DEX组在DEX干预后第1天至第3、7天动力蛋白激活蛋白的蛋白表达显著增加(P<0.05)。对照组干预后第3天至第7天动力蛋白激活蛋白的蛋白表达显著增加(P<0.05),而0.1 μmol/L DEX组干预后第3天至第7天动力蛋白激活蛋白的蛋白表达显著降低(P<0.05)。DEX干预后第3、7天,0.1 μmol/L DEX组和1.0 μmol/L DEX组大脑皮质神经元中动力蛋白激活蛋白的蛋白表达水平显著低于对照组(P<0.05)。DEX干预后第7天,1.0 μmol/L DEX组动力蛋白激活蛋白的蛋白表达水平显著低于0.1 μmol/L DEX组(P<0.05)。

结论

DEX可能以浓度和时间依赖的方式影响体外培养的胎鼠大脑皮质神经元中DHC和动力蛋白激活蛋白的蛋白表达。

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