Development of an in vitro screening method for idiosyncratic hepatotoxic components in traditional Chinese medicine: a case study with Epimedii Folium and Psoraleae Fructus / 药学学报

Idiosyncratic drug-induced liver injury (IDILI) has long posed a challenging and pivotal concern in pharmaceutical research. The complex composition of traditional Chinese medicine (TCM) has introduced a bottleneck in current research, hindering the elucidation of the component basis associated with IDILI in TCM. Using Epimedii Folium (EF) and Psoraleae Fructus (PF) as illustrative examples, this study endeavors to establish an in vitro evaluation model, providing a high-throughput and preliminary assessment method for screening components related to TCM-induced IDILI. A TNF-α-mediated HepG2 susceptible model was first established in this study, with the focus on the index components present in EF and PF. The release of lactate dehydrogenase (LDH) in the cell supernatant served as the detection index. A concentration-toxicity response curve was constructed, and the hepatotoxic components of EF and PF were identified utilizing the synergistic toxicity index. The LDH results unveiled the hepatotoxic effects of bavachin, backuchiol, isobavachin, neobavaisoflavone, psoralidin, isobavachalcone, icarisid I, and icarisid II on both normal and susceptible cells, categorizing these 8 components as both direct hepatotoxicity components and idiosyncratic hepatotoxicity components. Bavachin and neobavaisoflavone exhibited no hepatotoxicity on normal cells but demonstrated significant effects on susceptible cells, designating them as potential idiosyncratic susceptible hepatotoxicity components. The study further delineated that 10 EF components and 3 PF components were direct immune-promoting hepatotoxicity components. Additionally, 14 idiosyncratic immune-promoting hepatotoxicity components were identified, encompassing 10 EF components and 4 PF components, with neobavaisoflavone, bavachinin, and isobavachin being potential idiosyncratic susceptible immune-promoting hepatotoxicity components. Synergistic toxicity index results indicated that 13 idiosyncratic immune-promoting hepatotoxicity components (except anhydroicaritin) combined with bavachin demonstrated synergistic hepatotoxicity on susceptible cells. Notably, 3 idiosyncratic susceptible immune-promoting hepatotoxicity components combined with bavachin exhibited synergistic hepatotoxicity, with neobavaisoflavone displaying the highest synergistic toxicity index and bavachinin the lowest. In summary, this methodology successfully screens hepatotoxic and immune-promoting hepatotoxic components in EF and PF, distinguishing the types of components inducing hepatotoxicity, evaluating the hepatotoxicity degree of each component, and elucidating the synergistic relationships among them. Importantly, these findings align with the characteristics of IDILI. The method provides an effective model tool for the fundamental research of TCM-related IDILI components.

Effects of fructus psoraleae on specific hepatotoxicity in a state of immune stress / 中国药理学通报

Aim To investigate the hepatotoxic effect of aqueous extract of fructus psoraleae (WEFP) on lipopolysaccharide (LPS)-induced hepatotoxicity in SD rats under immune stress and its mechanism. Methods SD rats were divided into control (CON), LPS, WEFP, LPS+WEFP group. The LPS and LPS+WEFP groups were injected with 4 mg·kg-1 LPS via tail vein; 2 h later, the rats in WEFP group and LPS+WEFP group received the WEFP (1.1 g·kg-1·d-1) by oral gavage for seven consecutive days. Different endpoints such as body weight, liver index, bile flow rate, serum biochemical, histopathological changes, inflammatory cytokines, protein and mRNA expression levels were determined to clarify the liver toxicity and mechanism of WEFP. Results Compared with the CON group, rats in the LPS group had no significant changes in body weight, liver coefficient, serum ALT, AST, and ALP liver injury indicators; mild steatosis in the liver of the rats in the WEFP group did not cause liver damage; for rats in the LPS+WEFP group, body weight and bile excretion decreased, liver coefficient, serum ALT, AST, ALP, TBA levels significantly increased, and IL-1 and TNF-α secretion in the liver increased; at the same time, the pathological changes such as inflammatory reaction, cholestasis, and steatosis appeared in liver, RhoA mRNA and protein expression increased, and TLR4 and ICAM-1 pro-inflammatory gene expression increased, leading to acute liver injury. Conclusions The non-hepatotoxic dose of LPS can cause the same dose of psoralen to show more obvious liver toxicity, leading to the body's immunospecific response. Psoralen can cause immune stress rats to activate the expression of RhoA and other pro-inflammatory genes, further aggravate the release of inflammatory factors,and promote inflammatory reaction damage to liver cells and intrahepatic bile duct tissues,leading to obstruction of bile acid efflux and causing special effects such as heterogeneous liver injury.

Research progress in mechanisms of idiosyncratic drug-induced liver injury mediated by inflammatory stress / 药学学报

Idiosyncratic drug-induced liver injury (IDILI) is an unpredictable serious adverse drug reaction, which only occurs in a minority of special susceptible individuals. Although the mechanism of IDILI has not been fully understood, several hypotheses have been proposed to explain the action mode and specific mechanism of IDILI. Of these hypotheses, inflammatory stress hypothesis is one of the most important theories. Under the condition of inflammatory stress, drugs interact with inflammation and mediate the occurrence of IDILI through a variety of mechanisms, which can induce the production of inflammatory cytokines, activate coagulation system, affect the activity of metabolites, induce cholestasis, affect mitochondrial damage, and others. This review will summarize the main mechanisms and influencing factors of IDILI mediated by inflammatory stress, in order to provide a reference for preclinical drug development and basic research on drug-induced liver injury.

Bavachin enhances NLRP3 inflammasome activation induced by ATP or nigericin and causes idiosyncratic hepatotoxicity / 医学前沿

Psoraleae Fructus (PF) is a well-known traditional herbal medicine in China, and it is widely used for osteoporosis, vitiligo, and other diseases in clinical settings. However, liver injury caused by PF and its preparations has been frequently reported in recent years. Our previous studies have demonstrated that PF could cause idiosyncratic drug-induced liver injury (IDILI), but the mechanism underlying its hepatotoxicity remains unclear. This paper reports that bavachin isolated from PF enhances the specific stimuli-induced activation of the NLRP3 inflammasome and leads to hepatotoxicity. Bavachin boosts the secretion of IL-1β and caspase-1 caused by ATP or nigericin but not those induced by poly(I:C), monosodium urate crystal, or intracellular lipopolysaccharide. Bavachin does not affect AIM2 or NLRC4 inflammasome activation. Mechanistically, bavachin specifically increases the production of nigericin-induced mitochondrial reactive oxygen species among the most important upstream events in the activation of the NLRP3 inflammasome. Bavachin increases the levels of aspartate transaminase and alanine aminotransferase in serum and hepatocyte injury accompanied by the secretion of IL-1β via a mouse model of lipopolysaccharide-mediated susceptibility to IDILI. These results suggest that bavachin specifically enhances the ATP- or nigericin-induced activation of the NLRP3 inflammasome. Bavachin also potentially contributes to PF-induced idiosyncratic hepatotoxicity. Moreover, bavachin and PF should be evaded among patients with diseases linked to the ATP- or nigericin-mediated activation of the NLRP3 inflammasome, which may be a dangerous factor for liver injury.

The compound cell model-based evaluation for idiosyncratic liver injury of Cis-SG and Trans-SG / 药学学报

In this study, a composite cell model for evaluation of idiosyncratic drug-induced liver injury (IDILI) was established in vitro from the perspective of immune inflammation. And this model was used to evaluate the risk of IDILI for 2,3,5,4'-tetrahydroxy-cis-stilbene-2-O-β-glucoside (Cis-SG) and 2,3,5,4'-tetrahydroxy-trans-stilbene-2-O-β-glucoside (Trans-SG). To determine the low, medium, and high dosage of Cis-SG and Trans-SG, CellTiter-Glo® 3D Cell Viability Assay was used to detect the effects of Cis-SG and Trans-SG on cell viability of HepG2 cells in three dimensional (3D) culture, and MTT assay was used to detect the effects of Cis-SG and Trans-SG on cell viability of THP-1 derived macrophages. THP-1 derived macrophages were incubated by Cis-SG and Trans-SG directly or supernatants from HepG2 cells incubated with them. Enzyme linked immunosorbent assay (ELISA) was used to detect the levels of interleukin-1β (IL-1β) in the supernatants of the THP-1 derived macrophages. Western blot and reverse transcription-polymerase chain reaction (RT-PCR) were used to determine the expression of apoptosis-associated speck-like protein (ASC), Nod-like receptor protein 3 (NLRP3), cysteinyl aspartate specific proteinase-1 (caspase-1), and IL-1β in THP-1 derived macrophages. The results showed that there was no effect on the secretion of IL-1β in THP-1 derived macrophages incubated by Cis-SG and Trans-SG directly. However, the secretion of IL-1β, the protein and mRNA expression of ASC, NLRP3, caspase-1, and IL-1β significantly increased in THP-1 derived macrophages incubated by supernatants from HepG2 cells incubated with 1, 5, and 25 μmol·L-1 Cis-SG or 25 μmol·L-1 Trans-SG. In summary, the composite cell model for evaluation of IDILI established in vitro has been successfully applied in testing Cis-SG and Trans-SG. This composite cell model is helpful to evaluate and screen drugs with IDILI risk in vitro preliminarily, which provides methods for predicting and solving the idiosyncratic liver toxicity of drugs.

Discussion on influence factors, mechanism and traditional Chinese medicine pathogenesis of idiosyncratic drug-induced liver injury / 中国中药杂志

Idiosyncratic drug-induced liver injury (IDILI) is a kind of unique adverse drug reaction with relative high morbidity compared with other idiosyncratic diseases. Its occurrence, however, has nothing to do with pharmacological effects and clinical dosage of drugs administered, and only a small number of susceptible individuals will suffer from it. Especially to deserve to be mentioned, the proportion of TCM-induced IDILI showed an ascending trend year by year. So in this article, the author has reviewed some facts related with TCM-induced IDILI, including the predisposing causes and occurrence mechanism, and tries to provide reference for the prevention, diagnosis and treatment of TCM-induced IDILI through the analysis of characteristics and research status of TCM-induced IDILI and exploration of the internal relationship between Chinese medicine constitution type and IDILI.

Progress in research on the pathogenesis of immune regulation and idiosyncratic drug-induced liver injury / 药学学报

Idiosyncratic drug-induced liver injury(IDILI)is an adverse drug reaction that occurs only in a minority of the population. IDILI also has many characteristics such as unpredictable and low morbidity, its occurrence has often not been clearly correlated with the dose, route, or duration of drug administration. Several studies have shown that IDILI is a synergistic effect which was caused by body diathesis, environment and drugs. In addition, evidence also suggests that most IDILIs are mediated by immunity. Chemical medicines-related IDILIs have been extensively studied, and a variety of immunological mechanism hypotheses have also emerged to explain the pathogenesis and characteristics of chemical medicines-related IDILIs. However, the traditional Chinese medicine(TCM)-related IDILI has always been neglected due to the complexity and specificity of TCM. In recent years, TCM-related IDILI has been gradually confirmed by researchers, and formed a new hypothesis, a immunological stress-mediated tri-elements synergetic mechanism hypothesis, which can reveal the pathogenesis and clinical characteristics of TCM-related IDILI. This paper is prepared to summarize the immunological mechanism hypotheses of chemical medicine-related IDILI and TCM-related IDILI to provide a scientific basis for guiding IDILI research and establishing its clinical risk prevention and control measures.

Hepatotoxicidad por isoniazida en un paciente con enfermedad de Crohn: Reporte de caso y revisión de la literatura / Hepatotoxicity due to Isoniazide in a Patient with Crohn’s Disease: Case Report and Literature Review

La isoniazida se utiliza para el tratamiento o profilaxis de la tuberculosis; sin embargo, su uso puede asociarse con reacciones hepáticas adversas. La hepatitis clínicamente manifiesta sucede en 0,5%-1% de los pacientes que reciben isoniazida como monoterapia. En este artículo se describe el caso de un paciente con enfermedad de Crohn que cursó con hepatotoxicidad grave por isoniazida, y se hace una revisión de la literatura al respecto

Isoniazid is used for treatment or prophylaxis of tuberculosis but may be associated with adverse hepatic reactions. Clinically manifest hepatitis occurs in 0.5%-1% of patients who receive isoniazid as monotherapy. This article describes the case of a patient with Crohn’s disease who experienced severe hepatotoxicity due to isoniazid. It also reviews the literature.

Understanding of ACG Clinical Guideline:Diagnosis and Management of Idiosyncratic Drug-induced Liver Injury / 胃肠病学

The American College of Gastroenterology( ACG)released the first clinical guideline for drug-induced liver injury( DILI)in June,2014:the diagnosis and management of idiosyncratic drug-induced liver injury( IDILI), which includes all aspects of IDILI,such as risk factors,diagnosis,differential diagnosis and treatment. The guideline discusses especially the issues on herbal and dietary supplements( HDS)induced liver injury and DILI in patients with chronic liver disease. The guideline is of great value for standardizing the clinical diagnosis and treatment,providing the best recommendations for management of the patients with DILI. Since the evidence of research in the field of DILI is relatively weak at this moment,the clinicians should consider the possible individual difference of the patients under the frame of guideline for making the best clinical decision.