Table 1_Comprehensive safety assessment of Qiwei Tiexie Pill: integrating histopathological, biochemical, and metabolomic analyses in a rat model.docx

IntroductionQiwei Tiexie Pill (QWTX) is a Tibetan medicine formulation containing processed iron powder that requires systematic safety evaluation. This study aimed to assess the acute toxicological mechanisms of QWTX and its key mineral component, processed iron powder.
MethodsAn integrative approach combining histopathological examination, serum biochemistry, and multi-platform metabolomics (1H NMR and LC-MS) was employed to evaluate toxicological responses in Sprague-Dawley rats following 7-day oral administration.
ResultsWhile both treatments preserved hepatic structural integrity without inducing hepatotoxicity, significant renal effects were observed in a dose- and formulation-dependent manner. High-dose processed iron powder caused moderate renal histopathological alterations, primarily vascular changes and hemorrhage, alongside metabolic disruptions in both liver and kidney. QWTX, despite inducing significant metabolic perturbations at high dose, maintained normal renal architecture, indicating a protective effect conferred by its herbal components. Metabolomic and biochemical analyses revealed systemic metabolic reprogramming across four interdependent physiological domains: energy metabolism characterized by TCA cycle impairment and enhanced BCAA catabolism; oxidative stress evidenced by glutathione depletion and lipid peroxidation; nitrogen metabolism showing a “hepato-renal disconnect”; and neuroendocrine regulation with widespread hormone pathway dysregulation.
DiscussionQWTX demonstrated a superior safety profile compared to processed iron powder alone, particularly in preserving renal structure and mitigating iron‐associated nephrotoxicity. However, high‐dose QWTX still triggered significant oxidative and metabolic stress, underscoring the importance of dose optimization in clinical use. These findings provide a systems‐level understanding of the acute toxicological profile of QWTX and processed iron powder, supporting the traditional principles of herbal‐metal synergy in Tibetan medicine while highlighting the need for long-term safety studies to evaluate cumulative mineral exposure and chronic metabolic effects.

**引言** 七味铁屑丸（Qiwei Tiexie Pill, QWTX）是一款含有炮制铁粉的藏药制剂，需开展系统性安全性评价。本研究旨在阐明七味铁屑丸及其核心矿物组分——炮制铁粉的急性毒理作用机制。

**方法** 本研究采用组织病理学检查、血清生化检测及多平台代谢组学（1H NMR、LC-MS）联用的整合研究策略，对灌胃给药7天后的Sprague-Dawley大鼠的毒理应答进行评估。

**结果** 两种受试物均未破坏肝脏结构完整性，未诱发肝毒性，但均呈现出剂量与制剂依赖性的显著肾脏毒性效应。高剂量炮制铁粉可引发中度肾脏组织病理学改变，主要表现为血管病变与出血，并伴随肝脏与肾脏的代谢紊乱。尽管高剂量七味铁屑丸可引发显著代谢扰动，但其肾脏结构仍维持正常，提示其组方中的草药组分可发挥肾脏保护作用。代谢组学与生化分析结果显示，受试物可引发四个相互关联的生理通路层面的系统性代谢重编程：以三羧酸循环（TCA cycle）损伤、支链氨基酸（BCAA）分解代谢增强为特征的能量代谢紊乱；以谷胱甘肽耗竭、脂质过氧化为标志的氧化应激；表现为“肝肾代谢脱节”的氮代谢异常；以及伴随广泛激素通路失调的神经内分泌调控紊乱。

**讨论** 相较于单独使用炮制铁粉，七味铁屑丸展现出更优的安全性特征，尤其在维持肾脏结构完整、减轻铁相关性肾毒性方面效果显著。但高剂量七味铁屑丸仍可引发显著氧化应激与代谢应激，凸显了临床使用中优化给药剂量的重要性。本研究结果从系统层面阐明了七味铁屑丸与炮制铁粉的急性毒理学特征，既验证了藏药中“草药-矿物配伍协同”的传统理论，同时也提示需开展长期安全性研究，以评估矿物成分的累积暴露风险与慢性代谢效应。