丹参酮ⅡA减轻海马神经元细胞的放射性损伤

Tanshinone ⅡA protects hippocampal neuronal cells from radiation injury

目的探讨丹参酮ⅡA在体外对海马神经元细胞放射性损伤的影响,初步探讨其在放射性损伤保护过程中的相关机制。方法体外培养海马神经元细胞株HT-22,实验分为单纯对照组(Control)、单纯丹参酮ⅡA处理组(Tan)、照射组(RT)、照射+丹参酮ⅡA处理组(RT+Tan)以及照射+丹参酮ⅡA+糖酵解抑制剂2-脱氧-D-葡萄糖(2-deoxy-D-glucose,2-DG)处理组(RT+Tan+2-DG)。MTT法检测各组细胞的存活分数,流式细胞技术检测细胞凋亡,酶法检测上清液中乳酸含量,Western blot检测糖酵解相关酶LDHA的表达水平。结果照射后海马神经元细胞存活分数下降,而丹参酮ⅡA能提高放射线照射后海马神经元细胞的存活分数,并减少细胞凋亡。相对于单纯照射组,照射+丹参酮ⅡA处理组的海马神经元细胞HT-22的糖酵解相关酶LDHA表达上升,上清液乳酸含量明显增加。而加入糖酵解抑制剂2-DG后,放射处理后的海马神经元细胞LDHA表达下调,培养液上清液乳酸含量下降,且存活分数明显下降。结论丹参酮ⅡA能明显减轻放射线在体外对海马神经元细胞的放射损伤作用,其保护机制可能与调节放疗过程中糖酵解相关。

Objective To investigate the effects of tanshinone ⅡA on the radiation injury of hippocampal neurons in vitro and its mechanism in radiation injury. Methods The hippocampal neuron cell line HT - 22 was cultured in vitro. The experiment included four groups, control group （ Control）, irradiation group （ RT）, irradiation ＋ Tanshinone ⅡA treatment group （ RT ＋ Tan）, and irradiation ＋ Tanshinone ⅡA ＋ glycolysis inhibitor 2 - deoxy - D - glucose （2 - DG） treatment group （ RT ＋ Tan ＋ 2 - DG）. The cell survival fraction was detected by MTT assay. Apoptosis was detected by flow cytometry. The content of lactic acid in the supernatant of culture medium was determined by enzymatic method. The expression of glycolysis - related enzyme LDHA was assessed by Western blot. Results The survival fraction of hipp- ocampal neurons decreased after irradiation, while tanshinone ⅡA increased the survival fraction of hippocampal neurons and reduced the apoptosis rate after irradiation. The expression of glycolysis - related enzyme LDHA in hippocampal neu- ron HT - 22 cells treated with tanshinone ⅡA and irradiation treatment increased significantly compared with that of irradiated group. After adding the glycolysis inhibitor 2 - DG, the expression of LDHA in the hippoeampal neuron HT - 22 cells, the content of lactic acid in the supernatant and the survival fraction were significantly reduced. Conclusion Tanshinone ⅡA can significantly attenuate the radiation injury of hippocampal neurons in vitro, and its protective mechanism may be related to glycolysis during radiotherapy.