大黄肝毒性的精确亚细胞器靶向分析

Precise subcellular organelle-targeted analyses of the hepatotoxicity of rhubarb

[目的]药物性肝损伤(DILI)是急性肝功能衰竭的主要原因,对人类健康构成重大挑战。大黄(Rheumofficinale Baill,Da Huang)因其具有清热利尿的作用已应用于临床。然而,它对肝脏不同细胞器的毒性作用需要进一步验证。[方法]分析大黄肝毒性的潜在靶点及其与微粒体、线粒体、内质网(ER)、高尔基体(GA)和溶酶体等五种主要细胞器的潜在损伤关系。通过整合传统中药(ITCM)数据库、HERB数据库和网络药理学,我们分离和纯化了不同的细胞器,将它们与大黄的不同组分和单体在三磷酸腺苷(ATP)培养系统中孵育,并使用粒度分析和分子生物学实验检查了细胞器的结构和功能变化,以评估大黄是否会对肝脏主要细胞器造成损伤。[结果]通过虚拟预测和实验验证相结合,我们的研究证实,从大黄蒽醌中分离的大黄素、大黄鞣质中的儿茶素和大黄有机酸中的棕榈酸对代表性细胞器造成了最显著的功能和结构损伤。在所有单体化合物中,大黄素对微粒体、线粒体、内质网和溶酶体的损伤最大;儿茶素诱导微粒体和GA损伤;棕榈酸对肝脏微粒体和GA的损伤最大。表明大黄成分可能通过多细胞器损伤发挥肝毒性。[结论]我们的研究结果表明,大黄对不同的细胞器具有不同程度的损伤作用,进而通过损害细胞器的形态和功能来影响细胞的生命活动。本研究为大黄毒性成分和靶点的精细分析提供了理论依据和技术支持。

Objective:Drug-induced liver injury(DILI)is the leading cause of acute liver failure and poses a significant challenge to human health.Rhubarb(Rheum officinale Baill.DaHuang)has been clinically used for its heat-clearing and diuresis-promoting effects.However,its toxic effects on different organelles in the liver require further validation.Methods:We analyzed the potential targets affecting hepatotoxicity in rhubarb and the potential damage relationship with five major organelles,including microsomes,mitochondria,endoplasmic reticulum(ER),Golgi apparatus(GA),and lysosomes through Integrated Traditional Chinese Medicine(ITCM)/HERB databases and network pharmacology.We isolated and purified different organelles,incubated them with different fractions and monomers of rhubarb in an adenosine triphosphate(ATP)culture system and examined the structural and functional changes in the organelles using particle size analysis and molecular biological experiments to investigate whether rhubarb affects the damage and rupture of major organelles in the liver.Results:By combining virtual predictions and experimental verification,our research confirmed that emodin isolated from the anthraquinone of rhubarb,catechin in the tannins of rhubarb,and palmitic acid in the organic acids of rhubarb caused the most significant functional and structural damage to the representative organelles.Among all the monomeric compounds,emodin caused the most damage to the microsome,mitochondria,ER,and lysosome;catechin induced microsome and GA damage;and palmitic acid caused the most damage to microsomes and GA in the liver,suggesting that rhubarb components may exert hepatotoxicity through multi-organelle injury.Conclusions:Our findings revealed that rhubarb has varying degrees of damaging effects on different organelles,which in turn affects cellular life activities by impairing organelle morphology and function.This study provides a theoretical basis and technical support for a refined analysis of the toxic components and targets of rhubarb.