L-F001, a Multifunctional Fasudil-Lipoic Acid Dimer Prevents RSL3-Induced Ferroptosis via Maintaining Iron Homeostasis and Inhibiting JNK in HT22 Cells

Abstract

Ferroptosis, an iron-dependent form of non-apoptotic cell death, plays important roles in cerebral ischemia. Previously we have found that L-F001, a novel fasudil-lipoic acid dimer with good pharmacokinetic characters has good neuroprotection against toxin-induced cell death in vitro and in vivo. Here, we investigated the protective effects of L-F001 against a Glutathione peroxidase 4 (GPX4) inhibitor Ras-selective lethality 3 (RSL3) -induced ferroptosis in HT22 cells. We performed MTT, Transmission Electron Microscope (TEM), Western blot, and immunofluorescence analyses to determine the protective effects of L-F001 treatment. RSL3 treatment significantly reduced HT22 cell viability and L-F001 significantly protected RSL3-induced cell death in a concentration-dependent manner and significantly attenuated Mitochondrial shrinkage observed by TEM. Meanwhile, L-F001 significantly decreased RSL3-induced ROS and lipid peroxidation levels in HT22 cells. Moreover L-F001could restore GPX4 and glutamate-cysteine ligase modifier subunit (GCLM) levels, and significantly deceased Cyclooxygenase (COX-2) levels to rescue the lipid peroxidation imbalance. In addition, FerroOrange fluorescent probe and Western blot analysis revealed that L-F001 treatment decreased the total number of intracellular Fe2+ and restore Ferritin heavy chain 1 (FTH1) level in RSL3-induced HT22 cells. Finally, L-F001 could reduce RSL3-induced c-Jun N-terminal kinase (JNK) activation, which might be a potential drug target for LF-001. Considering that L-F001 has a good anti-ferroptosis effect, our results showed that L-F001 might be a multi-target agent for the therapy of ferroptosis-related diseases, such as cerebral ischemia.

SIRT1 activated by AROS sensitizes glioma cells to ferroptosis via induction of NAD+ depletion-dependent activation of ATF3

“…As the activators of SIRT1, both AMPK and JNK are not only activated by intracellular ROS, but also by RSL3. Previous reports showed that RSL3 triggered AMPK activation in human glioma U87 cells and activation JNK in mouse hippocampal HT22 Cells [ 23 , 55 ]. Thus, RSL3 might induce SIRT1 activation by influencing the activation of AMPK or JNK.…”

L‐F001, a multifunctional fasudil–lipoic acid dimer, antagonizes hypoxic–ischemic brain damage by inhibiting the TLR4/MyD88 signaling pathway

“…It has been shown to have a protective effect on a variety of neurodegenerative diseases (Bowerman et al., 2012 ). L‐F001, the combination of fasudil and the antioxidant lipoic acid, has been proved in vitro to be superior to fasudil and lipoic acid alone in the anti‐inflammatory and antioxidant effects (Chen et al., 2017 ; Luo et al., 2017 ; Peng et al., 2022 ; Shen et al., 2015 ). In this study, we further demonstrated in vivo that L‐F001 can reduce the expression of pro‐inflammatory cytokines and improve the structure and function of brain tissue by inhibiting the TLR4 pathway.…”

Study on the mechanism of zinc finger proteins ROCK1 and DMD in zinc-mediated cognitive impairment caused by aluminum exposure

“…However, it might have caused side effects such as intracranial hemorrhage, leukopenia, and damage to liver and kidney functions. The ROCK inhibitor L-F001 also exhibited effects of resisting oxidative stress, scavenging reactive oxygen species, and reducing intracellular glutathione levels, and it had the potential to emerge as a new drug for the treatment of Parkinson's disease and other neurodegenerative diseases (Peng et al, 2022). Nonetheless, its speci c effect still needed veri cation through large-scale data.…”