AMPK/STAT1通路参与土木香内酯改善H2O2引起的神经细胞PC12凋亡

doi:10.3969/j.issn.2095-9400.2019.11.006

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摘要目的：研究土木香内酯（ALA）对H2O2诱导神经细胞PC12凋亡的影响。方法：用不同浓度（1、5、10、20、50和100 μmol/L）的ALA处理细胞后进行MTT检测细胞增殖；用不同浓度（1和5 μmol/L）的ALA预先处理细胞后加入H2O2刺激细胞，进行流式和MTT检测细胞增殖情况；设立对照组、H2O2（100 μmol/L）组、ALA（1和5 μmol/L）组，ALA干预H2O2处理的细胞进行细胞流式检测细胞凋亡情况，MTT检测细胞增殖情况，Western blot检测AMPK和STAT1的磷酸化；利用siRNA沉默AMPK表达后再用ALA处理，Western blot检测STAT1的磷酸化的表达。结果：高浓度ALA抑制PC12细胞增殖；与对照组比，经H2O2刺激后PC12细胞凋亡增高；与H2O2组比较，ALA组细胞凋亡明显降低，并呈剂量依赖性；沉默AMPK表达不影响H2O2诱导PC12细胞STAT1的磷酸化表达；在沉默AMPK后再用ALA处理，与单沉默AMPK比较，STAT1的磷酸化的表达差异无统计学意义。结论：ALA可通过AMPK的激活调控H2O2诱导PC12细胞的STAT1的磷酸化表达，从而影响细胞凋亡。

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关键词 ：土木香内酯, PC12细胞, H2O2, AMPK, STAT1

Abstract：Objective: To study the effect of alantolactone (ALA) on apoptosis of PC12 induced by hydrogen peroxide (H2O2). Methods: Cell proliferation was detected by MTT after treatment with different concentrations of ALA (1, 5, 10, 20, 50, and 100 μmol/L). After cells were pretreated with different concentrations of ALA (1 and 5 μmol/L) and then stimulated by H2O2, cell proliferation was detected by flow cytometry and MTT. The control group, H2O2 (100 μmol/L) group and H2O2 (100 μmol/L)+ALA (1 and 5 μmol/L) were established. Cell apoptosis was detected by flow cytometry, cell proliferation by MTT and phosphorylation of AMPK and STAT1 by Western blot. Phosphorylation of STAT1 was detected by Western blot after treatment with small interfering RNA (siRNA) of AMPK and ALA in PC12. Results: High concentration of ALA inhibited the proliferation of PC12 cells. Compared with the control group, the apoptosis of PC12 cells increased after stimulation by H2O2. Compared with H2O2 group, the apoptosis of PC12 cells in H2O2+ALA group decreased significantly in a dose-dependent manner. Silencing AMPK expression did not affect H2O2-induced phosphorylation of STAT1 in PC12 cells induced by H2O2. ALA was used to treat PC12 cells after silencing AMPK. Compared with siAMPK+H2O2 group, phosphorylation of STAT1 in ALA+siAMPK+H2O2 group showed no significant difference. Conclusion: ALA could regulate the phosphorylation of STAT1 in PC12 cells induced by H2O2 via activating AMPK and thus affect cell apoptosis.

Key words： alantolactone PC12 cells H2O2 AMPK STAT1

收稿日期: 2019-02-12

基金资助:浙江省医药卫生科技计划项目（2019RC264）。

作者简介: 徐建飞（1986-），男，江西横峰人，主治医师，硕士。

链接本文:

https://xb.wmu.edu.cn/CN/10.3969/j.issn.2095-9400.2019.11.006 或 https://xb.wmu.edu.cn/CN/Y2019/V49/I11/814

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