Novel Application of Eupatilin for Effectively Attenuating Cisplatin-Induced Auditory Hair Cell Death via Mitochondrial Apoptosis Pathway

Abstract

Eupatilin (5,7-dihydroxy-3 ′ ,4 ′ ,6-trimethoxyflavone) is a pharmacologically active flavone that has been isolated from a variety of medicinal plants and possesses a number of pharmacological properties. This study evaluates the antioxidant and antiapoptotic effects of eupatilin on cisplatin-induced ototoxicity using in vitro and in vivo models including HEI-OC1 cells, cochlear hair cells, and zebrafish. Employing a CCK8 assay and Annexin V-FITC/PI double staining, we found that eupatilin significantly alleviated cisplatin-induced apoptosis and increased hair cell viability. The level of reactive oxygen species (ROS) was evaluated by CellROX green and MitoSOX Red staining. The results showed that eupatilin possesses antioxidant activity. MitoTracker Red staining indicated that eupatilin remarkably decreased mitochondrial damage. Furthermore, we demonstrated that eupatilin protects hair cells from cisplatin-induced damage. Mechanistic studies in cisplatin-induced HEI-OC1 cells revealed that eupatilin promoted Bcl-2 expression, downregulated Bax expression, reversed the increase in caspase-3 and PARP activity, and reduced the expression of phosphorylated p38 and JNK. Our data suggest a novel role for eupatilin as a protective agent against ototoxic drug-induced hair cell apoptosis by inhibiting ROS generation and modulating mitochondrial-related apoptosis.

Pretreatment with Eupatilin Attenuates Inflammation and Coagulation in Sepsis by Suppressing JAK2/STAT3 Signaling Pathway

“…19 It has antioxidant and anti-apoptotic effects against cisplatin-induced ototoxicity, as well as inhibiting inflammatory responses and aging in nucleus pulposus (NP) cells. 20,21 In addition, a derivative of Eupatilin to treat dry eye disease is being developed and is undergoing Phase I study. 22 Eupatilin can also reduce the inflammatory response in LPS-induced rats, decreasing the level of surfactant protein (SP)-A, SP-D, and inflammatory factors.…”

A genetically encoded tool to increase cellular NADH/NAD+ ratio in living cells

“…S3A-K ). In these experiments, superoxide was monitored using CellROX Green (localizes primarily to the nucleus and mitochondria) and mitoSOX (localizes primarily to the mitochondria); H 2 O 2 levels were probed with Amplex Red (27,28). We observed that expression of Ec STH and mito Ec STH did not impact the total cellular GSH/GSSG ratio ( Fig.…”

Cisplatin exposure acutely disrupts mitochondrial bioenergetics in the zebrafish lateral-line organ

“…The relationship between cisplatin and oxidative stress has been well established ( Gentilin et al, 2019 ; Kros and Steyger, 2019 ; Sheth et al, 2017 ), and prior studies demonstrate that mitochondria play a key role in cisplatin-induced oxidative stress in cancer and non-cancer cell lines ( Marullo et al, 2013 ) as well as hair cells ( Li et al, 2021 ; Lu et al, 2022 ). Here, we build on this literature and demonstrate that oxidative stress occurs shortly after cisplatin exposure.…”