夏枯草中的豆甾醇通过抑制炎症和铁死亡减轻脂多糖诱导的乳腺损伤。
Stigmasterol from Prunella vulgaris L. Alleviates LPS-induced mammary gland injury by inhibiting inflammation and ferroptosis.
作者信息
Sun Mingyang, Xu Dianwen, Liu Dianfeng, Ran Xin, Li Feng, Wang Jiaxin, Ge Yusong, Liu Yuhao, Guo Wenjin, Liu Juxiong, Cao Yu, Fu Shoupeng
机构信息
State Key Laboratory for Diagnosis and Treatment of Severe Zoonotic Infectious Diseases, Key Laboratory for Zoonosis Research of the Ministry of Education, Institute of Zoonosis, and College of Veterinary Medicine, Jilin University, Changchun 130062, China.
College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, Gansu, China.
出版信息
Phytomedicine. 2025 Feb;137:156362. doi: 10.1016/j.phymed.2025.156362. Epub 2025 Jan 3.
BACKGROUND
Dairy mastitis, a prevalent condition affecting dairy cattle, represents a significant challenge to both animal welfare and the quality of dairy products. However, current treatment options remain limited. Stigmasterol (ST) is a bioactive component of Prunella vulgaris L. (PV) with various pharmacological functions such as anti-inflammatory and anti-oxidation. At present, the specific effects and underlying mechanisms of PV and ST on dairy mastitis are still not fully understood.
PURPOSE
The aim of this research was to evaluate the pharmacological effects of PV and its active component ST on lipopolysaccharide (LPS) -stimulated bovine mammary epithelial cells (BMECs) and a mouse mastitis model, and to elucidate the possible mechanisms of action.
METHODS
UPLC-Q-TOF-MS/MS was employed to identify the constituents of PV. BMECs and mice were used to establish in vitro and in vivo models of mastitis. Western Blotting, RT-qPCR, immunofluorescence and other techniques were used to explore the effects of PV and ST on inflammatory factors, blood-milk barrier integrity, ferroptosis related indicators and their potential molecular mechanisms.
RESULTS
PV significantly attenuated the production of inflammatory mediators by LPS-stimulated BMECs. Subsequently, ST was found to be a potent anti-inflammatory agent in PV by inhibiting TLR4/NF-κB signaling pathway. This inhibition inhibits the myosin light chain (MLC)/MLC kinase signaling cascade and alleviates blood-milk barrier (BMB) disruption in BMECs. In addition, ferroptosis occurred in BMECs after LPS stimulation, and ST inhibited ferroptosis by stimulating Nrf2/GPX4 signaling pathway. Treatment of BMECs with the Nrf2 inhibitor ML385 significantly attenuated the therapeutic effect of ST. In vivo experiments further confirmed that both PV and ST attenuated LPS-induced breast tissue damage while reducing ferroptosis levels and restoring BMB.
CONCLUSION
ST from PV exhibits substantial anti-inflammatory properties and is a promising candidate for the treatment of dairy mastitis.
背景
奶牛乳腺炎是一种影响奶牛的常见病症，对动物福利和乳制品质量都构成了重大挑战。然而，目前的治疗选择仍然有限。豆甾醇（ST）是夏枯草（PV）的一种生物活性成分，具有多种药理功能，如抗炎和抗氧化作用。目前，PV和ST对奶牛乳腺炎的具体作用及潜在机制仍未完全明确。
目的
本研究旨在评估PV及其活性成分ST对脂多糖（LPS）刺激的牛乳腺上皮细胞（BMECs）和小鼠乳腺炎模型的药理作用，并阐明其可能的作用机制。
方法
采用超高效液相色谱-四极杆飞行时间串联质谱（UPLC-Q-TOF-MS/MS）鉴定PV的成分。利用BMECs和小鼠建立乳腺炎的体外和体内模型。采用蛋白质免疫印迹法、逆转录-定量聚合酶链反应（RT-qPCR）、免疫荧光等技术，探讨PV和ST对炎症因子、血乳屏障完整性、铁死亡相关指标的影响及其潜在分子机制。
结果
PV显著减轻了LPS刺激的BMECs中炎症介质的产生。随后发现，ST通过抑制TLR4/NF-κB信号通路，是PV中的一种强效抗炎剂。这种抑制作用抑制了肌球蛋白轻链（MLC）/MLC激酶信号级联反应，减轻了BMECs中的血乳屏障（BMB）破坏。此外，LPS刺激后BMECs发生铁死亡，而ST通过刺激Nrf2/GPX4信号通路抑制铁死亡。用Nrf2抑制剂ML385处理BMECs可显著减弱ST的治疗效果。体内实验进一步证实，PV和ST均减轻了LPS诱导的乳腺组织损伤，同时降低了铁死亡水平并恢复了BMB。
结论
PV中的ST具有显著的抗炎特性，是治疗奶牛乳腺炎的一个有前景的候选药物。
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