PI3K/GSK-3在LPA诱导PC12细胞凋亡中的作用

The role of PI3K/GSK-3 in Lysophosphatidic acid induced apoptosis of PC12 cells

目的探讨PI3K/GSK-3通路在溶血磷脂酸(LPA)诱导PC12细胞凋亡中的作用。方法培养PC12细胞,CCK8测定不同水平LPA处理下PC12细胞的细胞活力,Tunel法测定不同水平LPA处理下PC12细胞的细胞凋亡,测定PI3K、GSK-3抑制剂预处理对LPA诱导的PC12细胞活力和细胞凋亡的影响;Western Blot测定不同水平LPA处理下PC12细胞裂解液的凋亡蛋白酶Caspase 3水平,测定PI3K、GSK-3抑制剂预处理对LPA介导的PC12细胞裂解液Caspase 3水平变化的影响。结果 LPA剂量依赖导致PC12细胞活力下降,PI3K、GSK3α抑制剂能够减轻LPA介导的PC12细胞活力下降,GSK3β抑制剂能够促进LPA介导的PC12细胞活性损失;LPA以剂量依赖的方式诱导PC12细胞凋亡,PI3K、GSK3α抑制剂可减轻LPA介导的PC12细胞凋亡,GSK3β抑制剂可促进LPA介导的PC12细胞凋亡;LPA以剂量依赖的方式增加PC12细胞Caspase 3表达,PI3K、GSK3α抑制剂可抑制LPA介导的PC12细胞Caspase 3表达增加,GSK3β抑制剂可促进LPA介导的PC12细胞Caspase 3表达增加。结论 LPA/PI3K/GSK-3通路在神经细胞死亡的病理生理过程中起重要作用,干预LPA/PI3K/GSK-3通路将是一种潜在的创伤后治疗措施。

Objective To investigate the role of PI3 K/GSK-3 pathway in LPA-induced apoptosis of PC12 cells. Methods PC12 cells were exposed to different levels of Lysophosphatidic acid(LPA),the cell viability was determined with CCK8. Tunel assay was used to determine the apoptosis of PC12 cells treated with different levels of LPA. The cell viability and apoptosis of PC12 cells were assessed in the present of PI3 K and GSK-3 inhibitors. Western Blot was used to determine the levels of Caspase 3 in PC12 cells treated with different levels of LPA, and the change of Caspase 3 levels in PC12 cells was assessed in the present of PI3 K and GSK-3 inhibitors. Results LPA dose-dependently induced cell activity loss of PC12 cells. PI3 K and GSK3 alpha inhibitors could reduce the loss of cell viability induced by LPA. GSK3 beta inhibitor promoted the activity loss of PC12 cells induced by LPA. LPA induced PC12 cell apoptosis in a dose-dependent manner. PI3 K and GSK3 alpha inhibitors reduced LPA-mediated PC12 cell apoptosis. GSK3 beta inhibitor promoted LPA-mediated PC12 cell apoptosis.LPA increased the expression level of Caspase 3 in PC12 cells in a dose-dependent manner,PI3 K and GSK3 alpha inhibitors reduced the LPA-mediated increase of Caspase 3 expression level in PC12 cells, and GSK3 beta inhibitors promoted the LPA-mediated increase of Caspase 3 expression level in PC12 cells. Conclusion The LPA/PI3 K/GSK-3 pathway played an important role in the pathophysiological process of nerve cell death. Intervention of the LPA/PI3 K/GSK-3 pathway would be a potential post-traumatic therapy.