Icariside I specifically facilitates ATP or nigericin-induced NLRP3 inflammasome activation and causes idiosyncratic hepatotoxicity

Abstract

Background Epimedii Folium (EF) is commonly used for treating bone fractures and joint diseases, but the potential hepatotoxicity of EF limits its clinical application. Our previous study confirms that EF could lead to idiosyncratic drug-induced liver injury (IDILI) and hepatocyte apoptosis, but the mechanism remains unknown. Studies have shown that NLRP3 inflammasome plays an important role in the development of various inflammatory diseases such as IDILI. Specific stimulus-induced NLRP3 inflammasome activation may has been a key strategy for lead to liver injury. Therefore, main compounds derived from EF were chosen to test whether the ingredients in EF could activate the NLRP3 inflammasome and to induce IDILI. Methods Bone-marrow-derived macrophages (BMDMs) were treated with Icariside I, and then stimulated with inflammasome stimuli and assayed for the production of caspase-1 and interleukin 1β (IL-1β) and the release of lactate dehydrogenase (LDH). Determination of intracellular potassium, ASC oligomerization as well as reactive oxygen species (ROS) production were used to evaluate the stimulative mechanism of Icariside I on inflammasome activation. Mouse models of NLRP3 diseases were used to test whether Icariside I has hepatocyte apoptosis effects and promoted NLRP3 inflammasome activation in vivo. Results Icariside I specifically enhances NLRP3 inflammasome activation triggered by ATP or nigericin but not SiO2, poly(I:C) or cytosolic LPS. Additionally, Icariside I does not alter the activation of NLRC4 and AIM2 inflammasomes. Mechanically, Icariside I alone does not induce mitochondrial reactive oxygen species (mtROS), which is one of the critical upstream events of NLRP3 inflammasome activation; however, Icariside I increases mtROS production induced by ATP or nigericin but not SiO2. Importantly, Icariside I leads to liver injury and NLRP3 inflammasome activation in an LPS-mediated susceptibility mouse model of IDILI, but the effect of Icariside I is absent in the LPS-mediated mouse model pretreated with MCC950, which is used to mimic knockdown of NLRP3 inflammasome activation. Conclusions Our study reveals that Icariside I specifically facilitates ATP or nigericin-induced NLRP3 inflammasome activation and causes idiosyncratic hepatotoxicity. The findings suggest that Icariside I or EF should be avoided in patients with diseases related to ATP or nigericin-induced NLRP3 inflammasome activation, which may be risk factors for IDILI.

Astragaloside IV alleviates oxidative stress‑related damage via inhibiting NLRP3 inflammasome in a MAPK signaling dependent pathway in human lens epithelial cells

“…MtROS is one critical upstream event for regulating the activation of NLRP3 inflammasome, and mtROS also contributes to enhancing the activation of NLRP3 inflammasome through molecules such as Icariside I (Gao et al, 2021;. The oxidative status can be indicated by several markers, which are substrates for the reaction between ROS and biomolecules.…”

Epimedium koreanum Nakai–Induced Liver Injury—A Mechanistic Study Using Untargeted Metabolomics

“…In our study, it was also found that GPX4 and GSH decreased after EEE administration, which could not only induce ferroptosis in liver cells to produce direct toxicity, but also lead to non-specific liver injury by activating the NLRP3 inflammasome. According to literature reports, not only EEE can cause direct liver injury, but its main components such as icariside I and icariside II can also induce idiosyncratic liver injury (IDILI) by activating the NLRP3 inflammasome ( Wang Z et al, 2020 ; Gao et al, 2021 ). In addition, in the KEGG pathway analysis, we found that metabolic disturbance caused by EEE might also cause necroptosis in hepatocytes.…”

“…At present, the research on the hepatotoxicity of Epimedii Folium has been gradually carried out. Some researchers have found that Icariside I and Icariside II (the main components of Epimedii Folium) could induce specific hepatotoxicity by enhancing the activation of nlrp3 inflammasome ( Wang Z et al, 2020 ; Gao et al, 2021 ). Other researchers alsohave found that both 2″-O-rhamnosylicariside II and Sagittatoside B caused severe hepatocyte vacuolation and hepatocyte degeneration in adult zebrafish after 15 consecutive days of treatment ( Zhong et al, 2019 ).…”

An Integrative Pharmacology Based Analysis of Refined Liuweiwuling Against Liver Injury: A Novel Component Combination and Hepaprotective Mechanism

“…Protein extraction and western blotting assays on cell culture supernatant and whole cell lysis were performed as described previously ( Gao et al, 2021 ).…”