Meng Zhaoping, Zhang Wei, Guo Zixian, Wang Liyang, Liu Wenfei, Cao Ling, Zhang Yuhua, Meng Xiangcai
Department of Pharmacognosy, Heilongjiang University of Chinese Medicine, Harbin, Heilongjiang, China.
Heilongjiang Greater Hinggan Mountains Region Agriculture Forestry Research Institute, Jagdaqi, Heilongjiang, China.
PLoS One. 2025 Jul 18;20(7):e0327497. doi: 10.1371/journal.pone.0327497. eCollection 2025.
The fruits of Schisandra chinensis (Turcz.) Baill. (Schisandrae chinensis fructus) are a well-known herbal medicine, known for its hepatoprotective, antidepressant, antioxidant, and sedative-hypnotic properties. Over-exploitation of wild resources led to the rise of cultivation, along with a decrease in quality. Exposure of plants to adversity must generate substantial quantities of reactive oxygen species (ROS) and result in cellular damage. In response, secondary metabolites are produced to neutralize ROS; these secondary metabolites are usually the active ingredient of herbal medicine, so the quality of herbal medicine is closely related to the environment and ROS. The interplay of exogenous Nitric Oxide (NO, supplied as sodium nitroprusside) and high-temperature stress can simulate adversity and improve the quality of Schisandrae chinensis fructus; neverless, the underlying mechanism remains largely unexplored. In this study, we examined the changes in intracellular ROS levels as well as phenylalanine deaminase activities after stress and analyzed the metabolic changes using ultra performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MSE). The results showed that the level of superoxide anion (O2.-) and H2O2 increased by 25.8% and 331%, respectively, the activity of phenylalanine aminotransferase (PAL) by 69.3% on the 2nd day, indicating that combination of exogenous NO with high-temperature stress could lead to physiological states of adversity stress. Twenty-two differential metabolites (VIP ≥ 1) were identified using untargeted metabolomics. 3 primary metabolites, namely mannose, pyruvate, and shikimic acid, together with 2 secondary metabolites, phenylalanine and mevalonic acid, were observed to decrease. 17 secondary metabolites, including Schisandrin A, Schisandrin B, Schisandrin C, Gomisin D, Gomisin G, Gomisin H, Benzoylgomisin H, Benzoylgomisin O,Angeloylgomisin P, Catechin, Isorhamnetin, Luteolin, Cinnamic acid, Hydroxycinnamic acid, Hexahydrocurcumin, Coniferyl alcohol, Phenylalanine, Terpinolene and Mevalonic acid, exhibited increases in their levels by 10.64, 1.84, 1.40, 1.64, 4.46, 8.18, 1.72, 10.20, 2.08, 1.27, 1.57, 1.18, 2.01, 1.12, 1.88, 1.15, and 3.17-fold, respectively. Under stress conditions, intracellular ROS levels increased, and a significant portion of primary metabolites were used for the biosynthesis of secondary metabolites with higher antioxidant activity. This redistribution of metabolic flows from basal metabolism to secondary metabolism to defend against ROS. The combination of exogenous NO with high-temperature enhances secondary metabolism of Schisandra chinensis fruit, which opens new avenues for production of high-quality Schisandra chinensis fructus.
五味子果实是一种著名的草药，以其保肝、抗抑郁、抗氧化和镇静催眠特性而闻名。野生资源的过度开发导致了人工栽培的兴起，同时质量有所下降。植物暴露于逆境中必然会产生大量活性氧（ROS）并导致细胞损伤。作为响应，植物会产生次生代谢产物来中和ROS；这些次生代谢产物通常是草药的活性成分，因此草药的质量与环境和ROS密切相关。外源性一氧化氮（NO，以硝普钠形式提供）与高温胁迫的相互作用可以模拟逆境并提高五味子果实的质量；然而，其潜在机制在很大程度上仍未得到探索。在本研究中，我们检测了胁迫后细胞内ROS水平以及苯丙氨酸脱氨酶活性的变化，并使用超高效液相色谱-四极杆飞行时间串联质谱（UPLC-Q-TOF-MSE）分析了代谢变化。结果表明，超氧阴离子（O2.-）和H2O2水平分别增加了25.8%和331%，苯丙氨酸转氨酶（PAL）活性在第2天增加了69.3%，表明外源性NO与高温胁迫的组合可导致逆境胁迫的生理状态。使用非靶向代谢组学鉴定出22种差异代谢物（VIP≥1）。观察到3种初级代谢物，即甘露糖、丙酮酸和莽草酸，以及2种次生代谢物，苯丙氨酸和甲羟戊酸，含量降低。17种次生代谢物，包括五味子醇甲、五味子醇乙、五味子醇丙、戈米辛D、戈米辛G、戈米辛H、苯甲酰戈米辛H、苯甲酰戈米辛O、当归酰戈米辛P、儿茶素、异鼠李素、木犀草素、肉桂酸、羟基肉桂酸、六氢姜黄素、松柏醇、苯丙氨酸、萜品油烯和甲羟戊酸，其含量分别增加了10.64、1.84、1.40、1.64、4.46、8.18、1.72、10.20、2.08、1.27、1.57、1.18、2.01、1.12、1.88、1.15和3.17倍。在胁迫条件下，细胞内ROS水平升高，相当一部分初级代谢物被用于生物合成具有更高抗氧化活性的次生代谢物。这种代谢流从基础代谢向次生代谢的重新分配是为了抵御ROS。外源性NO与高温的组合增强了五味子果实的次生代谢，为高质量五味子果实的生产开辟了新途径。
