冬凌草甲素调节Hippo/YAP信号通路对氧糖剥夺复氧导致神经元损伤的影响*

作者：孔 明1，尚 颖1，李晓伟2，赵晓宝2，安 辉2

单位：1.保定市第二医院，河北 保定 071200；2.保定市第一中心医院，河北 保定 071200

引用：引用：孔明，尚颖，李晓伟，赵晓宝，安辉.冬凌草甲素调节Hippo/YAP信号通路对氧糖剥夺复氧导致神经元损伤的影响[J].中医药导报,2025,31(7):40-45.

DOI：10.13862/j.cn43-1446/r.2025.07.006

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摘要：目的：探讨冬凌草甲素（Ori）调节Hippo/YAP信号通路对氧糖剥夺复氧（OGD/R）导致的神经元损伤的影响。方法：使用不同浓度的Ori处理OGD/R诱导的HT22细胞。实验分为对照组（Control组）、OGD/R组、Ori低浓度（2 μmol/L）组（Ori-L组）、Ori中浓度（4 μmol/L）组（Ori-M组）、Ori高浓度（8 μmol/L）组（Ori-H组）和Ori高浓度（8 μmol/L）+YAP激活剂PY-60（10 μmol/L）组（Ori-H+PY-60组）。显微镜下观察细胞形态变化；Edu检测细胞增殖水平；流式细胞术检测细胞凋亡情况；采用酶联免疫吸附试验（ELISA）试剂盒测定乳酸脱氢酶（LDH）、过氧化氢酶（CAT）、超氧化物歧化酶（SOD）水平；活性氧（ROS）检测试剂盒测定ROS水平；免疫印迹法测定B细胞淋巴瘤-2（Bcl-2）、Bcl-2关联X蛋白（Bax）、Yes关联蛋白（YAP）、含PDZ结合基序的转录共激活因子（TAZ）蛋白表达。结果：与0 μmol/L Ori比较，2、4、8 μmol/L Ori处理后细胞活力显著增加（P<0.05），选择2、4、8 μmol/L的Ori进行后续实验。与Control组比较，OGD/R组细胞分布稀疏，细胞损伤严重，细胞出现皱缩肿胀，可见大量细胞死亡，Edu阳性率、CAT和SOD水平及Bcl-2水平显著降低（P<0.05），细胞凋亡率、LDH、ROS水平、Bax、YAP和TAZ水平显著增加（P<0.05）。与OGD/R组比较，Ori-L组、Ori-M组、Ori-H组细胞表现出较为正常的状态、细胞体完整，细胞损伤及皱缩肿胀程度减轻，死亡数量减少，Edu阳性率、CAT和SOD水平及Bcl-2水平显著增加（P<0.05），细胞凋亡率、LDH和ROS水平及Bax、YAP和TAZ水平显著降低（P<0.05），且呈浓度依赖性。YAP激活剂PY-60逆转了Ori对OGD/R诱导的神经元损伤的保护作用。结论：Ori可能通过调节Hippo/YAP信号通路改善OGD/R导致的神经元损伤。

关键词：神经元损伤；氧糖剥夺复氧；冬凌草甲素；Hippo/YAP信号通路

Abstract：

Objective: To investigate the effect of Oridonin (Ori) on neuronal damage induced by oxygen glucose deprivation/reoxygenation (OGD/R) by regulating the Hippo/YAP signaling pathway. Methods: OGD/R induced HT22 cells were treated with different concentrations of Ori. The experiment was separated into control group, OGD/R group, low concentration (2 μmol/L) Ori group (Ori-L group), medium concentration (4 μmol/L) Ori group (Ori-M group), high concentration (8 μmol/L) Ori group (Ori-H group) and Ori-H (10 μmol/L)+PY-60(10 μmol/L) group (Ori-H+PY-60 group). The morphological changes of cells was observed under microscope. Edu was applied to detect cell proliferation level. Flow cytometry was applied to detect cell apoptosis. Enzyme-linked immunosorbent assay (ELISA) were applied to measure lactate dehydrogenase (LDH), catalase (CAT) and superoxide dismutase (SOD) levels. Reactive oxygen (ROS) detection kit was applied to measure ROS level. Immunoblotting was applied to determine the expression of B cell lymphoma 2 (Bcl-2), Bcl-2 related X protein (Bax), Yes associated protein (YAP) and TAZ proteins. Results: Compared with 0 μmol/L Ori, the cell viability was significantly increased after treatment with 2, 4, and 8 μmol/L Ori (P<0.05), therefore, 2, 4, and 8 μmol/L Ori was selected for subsequent experiments. Compared with the control group, the OGD/R group showed sparse cell distribution, severe cell damage, shrinkage and swelling of cells, and obvious cell death, and the positive rate of Edu, the levels of CAT and SOD, and the level of Bcl-2 were obviously reduced in OGD/R group (P<0.05). The apoptosis rate, the levels of LDH and ROS, and the levels of Bax, YAP and TAZ were significantly increased in OGD/R group (P<0.05). Compared with the OGD/R group, the cells in the Ori-L group, Ori-M group and Ori-H group showed more normal state, intact cell bodies, reduced degree of cell damage and swelling, and reduced number of deaths, and the positive rate of Edu, the levels of CAT and SOD, and the level of Bcl-2 were significantly increased in Ori-L group, Ori-M group and Ori-H group(P<0.05). The apoptosis rate, the levels of LDH and ROS, and the levels of Bax, YAP, and TAZ were obviously reduced in Ori-L group, Ori-M group and Ori-H group (P<0.05). It was concentration dependent. The YAP activator, PY-60, reversed the protective effect of Ori on OGD/R induced neuronal damage. Conclusion: Ori may improve neuronal damage caused by OGD/R by regulating the Hippo/YAP signaling pathway.

Key words：neuron damage; oxygen glucose deprivation and reoxygenation; Oridonin; hippo/YAP signaling pathway

发布时间：2026-01-06

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