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目的研究不同浓度锰对大鼠神经细胞凋亡及内质网应激信号分子葡萄糖调节蛋白78(GRP78)和CCAAT增强子结合蛋白同源蛋白(CHOP)mRNA与蛋白以及天冬氨酸特异性半胱氨酸蛋白酶-12(caspase-12)蛋白表达的影响。方法以出生4~7 d的SPF级Wistar大鼠脑片为模型,分别用0(对照)、25、100、400μmol/L锰暴露24 h。检测培养液中乳酸脱氢酶(LDH)的释放量,神经细胞凋亡率以及神经细胞内GRP78、CHOP mRNA和蛋白及caspase-12蛋白的表达。结果与对照组比较,100和400μmol/L锰暴露组培养液中LDH的释放量均较高,而各浓度锰暴露组大鼠神经细胞凋亡率均较高,差异有统计学意义(P<0.01);且随着锰暴露浓度的升高,培养液中LDH的释放量和大鼠神经细胞凋亡率均呈逐渐升高的趋势。与对照组比较,100和400μmol/L锰暴露组神经细胞内GRP78 mRNA及各浓度锰暴露组大鼠神经细胞内CHOP mRNA的表达均较高,而100、400μmol/L锰暴露组神经细胞内GRP78和CHOP蛋白及各浓度锰暴露组大鼠神经细胞内caspase-12蛋白的表达也均较高,差异有统计学意义(P<0.01);且随着锰暴露浓度的升高,大鼠神经细胞内GRP78、CHOP mRNA和蛋白及caspase-12蛋白的表达均呈逐渐升高的趋势。结论锰可促使大鼠神经细胞凋亡及GRP78和CHOP表达升高。
Objective To investigate the effect of different concentrations of manganese on neuronal apoptosis and the expression of endoplasmic reticulum stress signaling molecule glucose regulated protein 78 (GRP78) and CCAAT enhancer binding protein homologous protein (CHOP) mRNA and protein aspartate specific half Effect of caspase-12 protein expression. Methods SPF Wistar rat brain slices from 4 to 7 days old were used as models and exposed to 0 (control) and 25, 100 and 400 μmol / L manganese for 24 h. The release of lactate dehydrogenase (LDH), the apoptosis rate of neurons and the expressions of GRP78, CHOP mRNA and protein and caspase-12 protein in nerve cells were detected. Results Compared with the control group, the release of LDH in the medium of 100 and 400 μmol / L manganese exposure was higher than that of the control group, while the apoptosis rate of nerve cells in each concentration of manganese exposed group was higher, the difference was statistically significant (P < 0.01). With the increase of manganese exposure, the release of LDH and the apoptosis rate of neurons in the culture medium tended to increase gradually. Compared with the control group, the expression of GRP78 mRNA in the neurons of 100 and 400μmol / L manganese exposure groups and the expression of CHOP mRNA in the neurons in each concentration of manganese exposure group were higher than that in the control group, while GRP78 And CHOP protein and the concentration of manganese exposed rats neuronal caspase-12 protein expression were also higher, the difference was statistically significant (P <0.01); and with the increase of manganese exposure, rat nerve cells The expression of GRP78, CHOP mRNA and protein and caspase-12 protein showed a gradually increasing trend. Conclusion Manganese can promote the apoptosis of nerve cells and the expression of GRP78 and CHOP in rats.