BCL2/BAX/ROS 通路参与黄芪甲苷对人脐静脉内皮细胞细胞焦亡的抑制作用。
The BCL2/BAX/ROS pathway is involved in the inhibitory effect of astragaloside IV on pyroptosis in human umbilical vein endothelial cells.
机构信息
Department of Critical Care Medicine, Foshan Hospital of Traditional Chinese Medicine, Guangzhou University of Chinese Medicine, Foshan, China.
The Second Clinical College of Guangzhou University of Chinese Medicine, Guangzhou, China.
出版信息
Pharm Biol. 2022 Dec;60(1):1812-1818. doi: 10.1080/13880209.2022.2101668.
CONTEXT
Astragaloside IV (AS-IV) is extracted from (Fisch.) Bunge (Fabaceae). However, its effects on endothelial cell injury remain unclear.
OBJECTIVE
To investigate the mechanisms underlying the effects of AS-IV on lipopolysaccharide (LPS)-induced endothelial injury .
MATERIALS AND METHODS
Human umbilical vein endothelial cells (HUVECs) were pre-treated with AS-IV (100 µmol/mL), 4-hydroxy-3-methoxyacetophenone (APO, 10 µmol/mL), -acetylcysteine (NAC, 50 µmol/mL) and Ac-YVAD-cmk (AC, 5 µmol/mL) for 2 h before 1 μg/mL LPS 24 h exposure. Untreated cells cultured without any exposure were used as controls. Cell viability, reactive oxygen species (ROS) and pyroptosis assays were performed. The pyroptosis related proteins were detected by western blot.
RESULTS
The rate in late pyroptosis (Q2-2) of AS-IV (13.65 ± 0.74%), APO (13.69 ± 0.67%) and NAC (15.87 ± 0.46%) groups was lower than the LPS group (21.89 ± 0.66%, < 0.05), while the rate in early pyroptosis (Q2-4) of AS-IV group (12.00 ± 0.26%) was lower than other groups ( < 0.05). The expression of NOX4, GSDMD, NLRP3, ASC and caspase-1 decreased after AS-IV, NAC or AC intervention ( < 0.05). The ROS production in AS-IV (4664 ± 153.20), APO (4094 ± 78.37), NAC (5103 ± 131.10) and AC (3994 ± 102.50) groups was lower than the LPS (5986 ± 127.30) group, while the mitochondrial BCL2/BAX protein expression ratio increased in AS-IV, APO and NAC groups ( < 0.05).
DISCUSSION AND CONCLUSIONS
AS-IV suppressed pyroptosis in LPS-activated HUVECs by inducing ROS/NLRP3-mediated inhibition of the inflammatory response, providing a scientific basis for clinical applications of AS-IV.
背景
黄芪甲苷（AS-IV）从蒙古黄芪（Fisch.）Bunge（豆科）中提取。然而，其对内皮细胞损伤的作用尚不清楚。
目的
探讨 AS-IV 对脂多糖（LPS）诱导的内皮细胞损伤的作用机制。
材料和方法
用人脐带静脉内皮细胞（HUVEC）进行实验，在 LPS 暴露 24 小时前，先用 AS-IV（100 μmol/mL）、4-羟基-3-甲氧基苯乙酮（APO，10 μmol/mL）、N-乙酰半胱氨酸（NAC，50 μmol/mL）和 Ac-YVAD-cmk（AC，5 μmol/mL）预处理细胞 2 小时。未暴露于任何物质的细胞作为对照。进行细胞活力、活性氧（ROS）和细胞焦亡检测。用 Western blot 检测细胞焦亡相关蛋白。
结果
AS-IV（13.65±0.74%）、APO（13.69±0.67%）和 NAC（15.87±0.46%）组的晚期细胞焦亡率（Q2-2）低于 LPS 组（21.89±0.66%， <0.05），而 AS-IV 组的早期细胞焦亡率（Q2-4）（12.00±0.26%）低于其他组（ <0.05）。AS-IV、NAC 或 AC 干预后，NOX4、GSDMD、NLRP3、ASC 和 caspase-1 的表达下降（ <0.05）。AS-IV（4664±153.20）、APO（4094±78.37）、NAC（5103±131.10）和 AC（3994±102.50）组的 ROS 生成量低于 LPS（5986±127.30）组，而 AS-IV、APO 和 NAC 组的线粒体 BCL2/BAX 蛋白表达比值增加（ <0.05）。
讨论与结论
AS-IV 通过诱导 ROS/NLRP3 介导的炎症反应抑制，抑制 LPS 激活的 HUVEC 中的细胞焦亡，为 AS-IV 的临床应用提供了科学依据。
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