黄芪总苷通过抑制P38通路缓解氧糖剥夺/复氧后的星形胶质细胞水肿并降低AQP4表达

doi:10.12122/j.issn.1673-4254.2026.02.06

摘要/Abstract

摘要：

目的探讨黄芪总苷（AST）对氧糖剥夺/复氧（OGD/R）诱导的星形胶质细胞肿胀的影响及其潜在分子机制。方法取2 d龄SD大鼠的乳鼠，共48只，随机分为4组，12只/组。从乳鼠大脑中提取原代星形胶质细胞，经OGD/R诱导损伤后，分别给予AST或AST联合P38激动剂U-46619处理。实验设置对照组、OGD/R组、AST组以及AST+U-46619组。其中，AST组在OGD/R处理前及过程中用AST处理细胞；AST+U-46619组在OGD/R处理前及过程中用AST和U-46619共同处理细胞。通过细胞体积检测、形态观察、活性氧物质（ROS）检测和乳酸脱氢酶（LDH）活性测定评估各组细胞的肿胀、ROS水平和损伤程度；采用免疫荧光共聚焦显微镜显示细胞膜上水通道蛋白4（AQP4）的蛋白表达情况；Western blotting检测P38信号通路的磷酸化水平及细胞膜上AQP4蛋白的表达水平。结果与OGD/R组相比，AST处理降低了OGD/R损伤引起的星形胶质细胞体积增加（P<0.05）、ROS水平（P<0.05）和LDH释放量（P<0.05），改善了细胞形态变化，抑制了细胞膜上AQP4的过度聚集，降低了P38的磷酸化水平（P<0.05）和细胞膜上AQP4蛋白表达（P<0.05）。然而，U-46619与AST联合处理可逆转AST对细胞体积、形态、LDH水平、P38磷酸化和AQP4表达水平的影响（均P<0.05）。结论 AST缓解OGD/R后星形胶质细胞水肿，其分子机制可能为抑制细胞中ROS产生和抑制P38信号通路激活，从而降低了AQP4在细胞膜上过度表达。

关键词: 黄芪总苷, 星形胶质细胞肿胀, 水通道蛋白4, P38, 氧糖剥夺/复氧

Abstract:

Objective To investigate the effect of total Astragalus saponins (AST) on astrocyte swelling induced by glucose-oxygen deprivation and reoxygenation (OGD/R) and its molecular mechanism. Methods Primary astrocytes isolated from the brain of post-natal day 2 SD rats were treated with solvent vehicle, AST, or AST combined with U-46619 (a p38 activator) 24 h prior to and during exposure to OGD/R. The changes in cell volume and morphology were observed, and intracellular reactive oxygen species (ROS) levels and lactate dehydrogenase (LDH) activity were detected. The expression of aquaporin 4 (AQP4) on cell membrane was observed using immunofluorescence confocal microscopy, and Western blotting was performed to detect the changes in p38 signaling pathway phosphorylation levels and AQP4 expression level. Results AST treatment significantly reduced OGD/R-induced increase in astrocyte volume, ROS levels and LDH release. The AST-treated cells also exhibited improved cell morphology, reduced aggregation of AQP4 on the cell membrane, and decreased p38 phosphorylation and AQP4 expression levels. The protective effects of AST against OGD/R-induced cell injuries were significantly attenuated by combined treatment of the cells with U-46619. Conclusion AST alleviate astrocyte swelling induced by OGD/R injury by inhibiting the activation of the p38 signaling pathway and AQP4 overexpression on the cell membrane.

Key words: total Astragalus glycosides, astrocyte swelling, aquaporin 4, p38 signaling pathway, oxygen-glucose deprivation/reoxygenation

尹钦, 雷雨逍, 王晓澍, 杨刚, 唐兆华. 黄芪总苷通过抑制P38通路缓解氧糖剥夺/复氧后的星形胶质细胞水肿并降低AQP4表达[J]. 南方医科大学学报, 2026, 46(2): 286-292.

Qin YIN, Yuxiao LEI, Xiaoshu WANG, Gang YANG, Zhaohua TANG. Total Astragalus saponins reduce astrocyte swelling and AQP4 expression following oxygen-glucose deprivation and reoxygenation by inhibiting the p38 pathway[J]. Journal of Southern Medical University, 2026, 46(2): 286-292.

图/表 5

图1 AST和U-46619对OGD/R后星形胶质细胞肿胀的影响

Fig.1 Effects of total Astragalus saponins (AST) and U-46619 on astrocyte swelling following glucose-oxygen deprivation and reoxygenation (OGD/R). A: Effect of AST on astrocyte volume following OGD/R (n=5). B: Effect of AST on morphology of astrocytes following OGD/R (Original magnification: ×200). #P<0.05 vs control group, *P<0.05 vs OGD/R group, &P<0.05 vs AST group.

图2 AST和U-46619对OGD/R后星形胶质细胞中ROS水平的影响

Fig.2 Effects of AST and U-46619 on intracellular ROS level in astrocytes following OGD/R (n=3, ×200). #P<0.05 vs control group; *P<0.05 vs OGD/R group.

图3 AST和U-46619对OGD/R后星形胶质细胞损伤的影响

Fig.3 Effects of AST and U-46619 on astrocyte injury following OGD/R (n=5). #P<0.05 vs control group; *P<0.05 vs OGD/R group; &P<0.05 vs AST group.

图4 AST和U-46619对OGD/R后星形胶质细胞P38通路的磷酸化的影响

Fig.4 Effects of AST and U-46619 on phosphorylation of the p38 pathway in rat astrocytes following OGD/R (n=5). p-P38: Phosphorylated P38. #P<0.05 vs control group; *P<0.05 vs OGD/R group; &P<0.05 vs AST group.

图5 AST和U-46619对OGD/R后星形胶质细胞膜上AQP4蛋白的聚集的影响。

Fig.5 Effects of AST and U-46619 on AQP4 protein aggregation on astrocyte membrane following OGD/R. A: Aggregation of AQP4 protein on astrocyte membrane in each group assessed by immunofluorescence staining (n=5, ×200). B: Expression of AQP4 protein on astrocyte membrane (m-AQP4) in each group assessed by Western blotting (n=5). #P<0.05 vs control group; *P<0.05 vs OGD/R group; &P<0.05 vs AST group.

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