Tenuifoliside A基于ERK通路保护皮质酮损伤C6细胞作用研究

Tenuifoliside A inhibits corticosterone-induced neurotoxicity of rat glioma cells via ERK pathway

目的探讨远志寡糖酯成分Tenuifoliside A(TFSA)对皮质酮诱导大鼠神经胶质瘤细胞(C6)损伤的保护作用及其分子机制。方法建立以0.8 mmol.L-1的皮质酮作用于C6细胞48h的损伤模型;用皮质酮预处理C6细胞30 min后加入30、10和3μmol.L-1的TFSA,共同作用48 h,CCK试剂盒检测TFSA对皮质酮诱导C6细胞损伤的存活率;荧光显微镜观察C6细胞Hoechst 33258染核后凋亡形态变化;Western blot检测C6细胞内ERK及凋亡相关蛋白Bcl-2和Bax的表达;Caspase检测试剂盒检测C6细胞内Caspase激酶活力的变化。结果与正常对照组比,0.8 mmol.L-1的皮质酮对C6细胞有明显的损伤作用,其细胞存活率降低至69.58%(与正常组相比,P<0.01);细胞核形态改变,出现凋亡小体;胞内ERK蛋白和抗凋亡蛋白Bcl-2表达量明显降低(与正常组相比,P<0.01),Caspase-3激酶的活力增加(P<0.01)。与模型组相比,各剂量组的TFSA均有不同程度对抗皮质酮损伤的作用,细胞存活率明显提高至84.48%(与模型组相比,P<0.01),同时细胞核形态有明显改善;细胞内的ERK蛋白和抗凋亡蛋白Bcl-2表达量明显增加(P<0.01);Caspase-3激酶活力明显降低。结论 TFSA对皮质酮诱导损伤的C6细胞具有保护作用,其机制可能与TFSA能激活C6细胞内ERK通路,提高抗凋亡蛋白Bcl-2表达,降低Caspase-3激酶活性有关。

Aim To investigate the neuroprotective effects of Tenuifoliside A（TFSA） on corticosterone-induced injury in rat glioma cells and explore the potential mechanism.Methods The injury model was established by treating C6 cells with 0.8 mmol·L-1 of corticosterone（CORT） by 48h;C6 cells were pretreated with CORT for 30 min followed by co-treated with 30,10 and 3 μmol·L-1 of TFSA for 48 h.Cell viability was detemined by Cell Counting Kit（CCK） assay;Hoechst 33258 staining was used to identify the neuroprotective effects of TFSA against neurotoxicity induced by CORT;Western blot was used to detect the expression of ERK,Bcl-2 and Bax protein.Caspase-3 activity was as a specific apoptotic marker in C6 cells exposed to CORT alone or co-treated with TFSA was also assayed.Results Compared with normal control group,CORT significantly decreased the viablity of C6 cells to 69.58%（P＆lt;0.01）.Western blot results showed that the protein levels of ERK and Bcl-2 decreased significantly.In contrast,the expression of Bax and the activity of Caspase-3 increased markedly（P＆lt;0.01）.However,TFSA could increase the viability of C6 cells damaged by CORT up to 84.48%（P＆lt;0.01）.Moreover,TFSA increased the protein level of ERK,Bcl-2 and decreased the protein level of Bax and the activity of Caspase-3 in C6 cells exposed to CORT（P＆lt;0.01）.Conclusion TFSA protects C6 cells from CORT-induced damage,and the neuroprotective mechanism may be associated with the activation of Bcl-2 and suppressing Bax and the activity of Caspase-3 via ERK pathway.