School of Public Health, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, China.
State Key Laboratory of Traditional Chinese Medicine Resources in Southwest China, Chengdu 611137, China.
Phytomedicine. 2024 Sep;132:155849. doi: 10.1016/j.phymed.2024.155849. Epub 2024 Jun 28.
With the increasing awareness of the safety of traditional Chinese medicine and food, as well as in-depth studies on the pharmacological activity and toxicity of Zanthoxylum armatum DC. (ZADC), it has been found that ZADC is hepatotoxic. However, the toxic substance basis and mechanism of action have not been fully elucidated. Hydroxy-α-sanshool (HAS) belongs to an amide compound in the fruits of ZADC, which may be hepatotoxic. However, the specific effects of HAS, including liver toxicity, are unclear.
The objectives of this research was to determine how HAS affects hepatic lipid metabolism, identify the mechanism underlying the accumulation of liver lipids by HAS, and offer assurances on the safe administration of HAS.
An in vivo experiment was performed by gavaging C57 BL/6 J mice with various dosages of HAS (5, 10, and 20 mg/kg). Biochemical indexes were measured, and histological analysis was performed to evaluate HAS hepatotoxicity. Hepatic lipid levels were determined using lipid indices and oil red O (ORO) staining. Intracellular lipid content were determined by biochemical analyses and ORO staining after treating HepG2 cells with different concentrations of HAS in vitro. Mitochondrial membrane potential, respiratory chain complex enzymes, and ATP levels were assessed by fluorescence labeling of mitochondria. The levels of proteins involved in lipogenesis and catabolism were determined using Western blotting.
Mice in the HAS group had elevated alanine and aspartate aminotransferase blood levels as well as increased liver index compared with the controls. The pathological findings showed hepatocellular necrosis. Serum and liver levels of triglycerides, total cholesterol, and low-density lipoprotein cholesterol levels were increased, whereas high-density lipoprotein cholesterol levels decreased. The ORO staining findings demonstrated elevated liver lipid levels. In vitro experiments demonstrated a notable elevation in triglyceride and total cholesterol levels in the HAS group. ATP, respiratory chain complex enzyme gene expression, mitochondrial membrane potential, and mitochondrial number were reduced in the HAS group. The levels of lipid synthesis-associated proteins (ACC, FASN, and SREBP-1c) were increased, and lipid catabolism-associated protein levels (PPARα and CPT1) and the p-AMPK/AMPK ratio were decreased in vivo and in vitro.
HAS has hepatotoxic effects, which can induce fatty acid synthesis and mitochondrial function damage by inhibiting the AMPK signaling pathway, resulting in aberrant lipid increases.
随着人们对中药和食品安全性的认识不断提高，以及对花椒（Zanthoxylum armatum DC.，ZADC）的药理活性和毒性的深入研究，发现 ZADC 具有肝毒性。然而，其毒性物质基础和作用机制尚未完全阐明。羟基-α-山椒素（HAS）属于花椒果实中的酰胺类化合物，可能具有肝毒性。然而，HAS 的具体作用，包括肝毒性，尚不清楚。
本研究旨在探讨 HAS 如何影响肝脂质代谢，阐明 HAS 引起肝脂质堆积的作用机制，为 HAS 的安全使用提供依据。
通过灌胃给予 C57BL/6 J 小鼠不同剂量 HAS（5、10 和 20mg/kg）进行体内实验。测定生化指标，进行组织学分析，评估 HAS 的肝毒性。通过脂质指数和油红 O（ORO）染色测定肝脂质水平。通过不同浓度 HAS 体外处理 HepG2 细胞，测定细胞内脂质含量和 ORO 染色。通过荧光标记线粒体测定线粒体膜电位、呼吸链复合体酶和 ATP 水平。通过 Western blot 测定参与脂生成和分解的蛋白质水平。
与对照组相比，HAS 组小鼠的丙氨酸氨基转移酶和天冬氨酸氨基转移酶血液水平升高，肝指数增加。病理检查显示肝细胞坏死。血清和肝脏中甘油三酯、总胆固醇和低密度脂蛋白胆固醇水平升高，而高密度脂蛋白胆固醇水平降低。ORO 染色结果显示肝脂质水平升高。体外实验显示 HAS 组甘油三酯和总胆固醇水平显著升高。HAS 组的 ATP、呼吸链复合体酶基因表达、线粒体膜电位和线粒体数量减少。HAS 组脂质合成相关蛋白（ACC、FASN 和 SREBP-1c）水平升高，体内和体外的脂质分解相关蛋白（PPARα 和 CPT1）和 p-AMPK/AMPK 比值降低。
HAS 具有肝毒性作用，可通过抑制 AMPK 信号通路诱导脂肪酸合成和线粒体功能损伤，导致脂质异常增加。
