Dong Jiaqi, Ma Qian, Yang Rong, Zhang Xiaosong, Hua Yongli, Ji Peng, Yao Wanling, Yuan Ziwen, Wei Yanming
College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China.
College of Veterinary Medicine, China Agricultural University, Beijing 100091, China.
Antioxidants (Basel). 2025 Sep 22;14(9):1140. doi: 10.3390/antiox14091140.
Heat stress (HS), a pervasive environmental stressor, significantly disrupts systemic physiological homeostasis, posing substantial threats to human and animal health. Sheng Mai San (SMS), a classic Traditional Chinese Medicine (TCM) formula, exerts its therapeutic effects by replenishing qi (the vital energy governing physiological functions) and nourishing yin (the material basis responsible for moistening and cooling actions). This formula demonstrates significant efficacy in astringing sweating and preventing collapse. However, its precise molecular mechanisms against HS-induced myocardial injury remain incompletely elucidated. This study initially employed physicochemical analytical methods to determine the contents of total polysaccharides, saponins, and flavonoids in SMS and evaluated its antioxidant activity. Subsequently, both in vitro and in vivo rat models of HS were established to systematically assess the alterations in reactive oxygen species (ROS), antioxidant enzymes (GSH, SOD, CAT), and heat shock proteins (HSP70, HSP90) following SMS intervention, thereby investigating HS-induced myocardial injury and the protective effects of SMS. Furthermore, Western blot, immunofluorescence, and qRT-PCR techniques were utilized to quantitatively analyze key molecules in the Keap1-Nrf2-HO-1 and Stub1-HSF1 signaling pathways. The results demonstrated that total polysaccharides were the most concentrated in SMS, followed by total saponins. This formula exhibited potent free radical scavenging capacity against DPPH, ABTS, and OH, along with significant reducing activity. HS-induced myocardial injury reached its peak severity at 6-12 h post-stress exposure. SMS intervention effectively suppressed excessive ROS generation, enhanced the activities of antioxidant enzymes (GSH, SOD, and CAT), and downregulated HSP70 and HSP90 mRNA expression levels, thereby significantly mitigating cardiomyocyte damage. Mechanistic investigations revealed that SMS conferred cardioprotection through dual modulation of the Keap1-Nrf2-HO-1 and Stub1-HSF1 signaling pathways. This study not only provides a novel TCM-based therapeutic strategy for preventing and treating HS-related cardiovascular disorders but also establishes a crucial theoretical foundation for further exploration of SMS's pharmacological mechanisms and clinical applications.
热应激（HS）是一种普遍存在的环境应激源，会严重破坏全身生理稳态，对人类和动物健康构成重大威胁。生脉散（SMS）是一种经典的中药方剂，通过补气（主导生理功能的 vital energy）和滋阴（负责滋润和清凉作用的物质基础）发挥治疗作用。该方剂在敛汗和防止虚脱方面具有显著疗效。然而，其对抗 HS 诱导的心肌损伤的确切分子机制仍未完全阐明。本研究首先采用物理化学分析方法测定 SMS 中总多糖、皂苷和黄酮类化合物的含量，并评估其抗氧化活性。随后，建立体外和体内 HS 大鼠模型，系统评估 SMS 干预后活性氧（ROS）、抗氧化酶（GSH、SOD、CAT）和热休克蛋白（HSP70、HSP90）的变化，从而研究 HS 诱导的心肌损伤及 SMS 的保护作用。此外，利用蛋白质免疫印迹、免疫荧光和 qRT-PCR 技术定量分析 Keap1-Nrf2-HO-1 和 Stub1-HSF1 信号通路中的关键分子。结果表明，总多糖在 SMS 中含量最高，其次是总皂苷。该方剂对 DPPH、ABTS 和 OH 具有强大的自由基清除能力，同时具有显著的还原活性。HS 诱导的心肌损伤在应激暴露后 6 - 12 小时达到最严重程度。SMS 干预有效抑制了过量 ROS 的产生，增强了抗氧化酶（GSH、SOD 和 CAT）的活性，并下调了 HSP70 和 HSP90 mRNA 的表达水平，从而显著减轻心肌细胞损伤。机制研究表明，SMS 通过对 Keap1-Nrf2-HO-1 和 Stub1-HSF1 信号通路的双重调节发挥心脏保护作用。本研究不仅为预防和治疗与 HS 相关的心血管疾病提供了一种基于中药的新型治疗策略，也为进一步探索 SMS 的药理机制和临床应用奠定了关键的理论基础。
