丹参酮ⅡA通过Nrf2调节ALDOC减轻海马神经元放射性损伤的机制

Tanshinone ⅡA regulates ALDOC to alleviate radiation injury in hippocampal neurons

目的探索丹参酮ⅡA通过Nrf2调控ALDOC的表达,从而促进海马神经元放射性损伤保护作用的分子机制。方法选择海马神经元细胞株HT 22为研究对象,予分组处理:对照组(Control)/单纯丹参酮ⅡA处理组(Tan)/单纯照射组(RT)/照射+丹参酮ⅡA处理组(RT+Tan),检测各组细胞ALDOCmRNA及蛋白水平的表达情况。构建ALDOC稳定过表达及干扰的细胞株,检测ALDOC的表达水平对糖酵解相关酶及自噬相关蛋白表达的影响。丹参酮ⅡA处理HT 22后检测Nrf2及ALDOC的表达水平,siRNA干扰细胞株Nrf2表达,再次检测其ALDOC表达水平。利用免疫荧光检测丹参酮ⅡA联合放射线照射后HT 22细胞Nrf2的变化。结果与对照组相比,RT+Tan组HT-22细胞ALDOC的mRNA及蛋白表达水平均出现明显上调。过表达ALDOC的细胞株,射线照射后的细胞存活率显著提高(F=126.13,P<0.05),凋亡比例降低(F=98.677,P<0.05);干扰ALDOC的表达,细胞则出现了相反的变化。ALDOC能增加经放射线照射后的HT 22细胞糖酵解,且促进自噬相关蛋白的表达。丹参酮ⅡA处理后,Nrf2及ALDOC表达明显上调,而抑制Nrf2后,ALDOC表达下降,免疫荧光显示丹参酮ⅡA促进Nrf2由细胞浆转移至细胞核。结论丹参酮ⅡA可能是通过Nrf2调节糖酵解酶ALDOC的表达改变从而调控糖酵解及自噬,对海马神经元放射性损伤起到保护作用。

Objective To explore the molecular mechanism of the protective effect of tanshinone ⅡA on radiation injury of hippocampal neurons HT-22 cells.Methods The hippocampal neuron cell line HT-22 was cultured in vitro,and the cells were irradiated with radiation and treated with tanshinone ⅡA.The survival fraction of the cells was detected by MTT assay,apoptosis was detected by flow cytometry,and ALDOC gene expression was detected by qRT-PCR.Western Blot was used to detect glycolysis-related enzymes and autophagy-related proteins.Results The expression of ALDOC,a cell glycolysis-related enzyme,was also significantly up-regulated at mRNA and protein levels.Overexpression of ALDOC significantly increased the survival rate of hippocampal neurons HT-22 cells after radiation exposure(F = 126.13,P < 0.05),and decreased the proportion of apoptosis(F = 98.677,P<0.05).Conversely,interference with ALDOC could significantly inhibit the cell viability of HT-22 cells after radiation exposure.ALDOC can increase the glycolysis of HT-22 cells after irradiation and promote the expression of autophagy-related proteins.Conclusion Tanshinone ⅡA may regulate the glycolysis and autophagy by regulating the expression change of the glycolytic enzyme ALDOC,and protect the radioactive injury of hippocampal neurons.