Gallic Acid Inhibits Mesaconitine-Activated TRPV1-Channel-Induced Cardiotoxicity

Abstract

Aconiti Kusnezoffii Radix (Caowu) is often combined or processed with Chebulae Fructus (Hezi) to achieve attenuation purposes in Mongolian medicine. Mesaconitine (MA), a main bioactive ingredient of Caowu, is also famous for its high cardiotoxicity while exerting good anti-inflammatory and analgesic properties. Gallic acid (GA), one of the leading chemical components in Hezi, possesses cardiac protection. This study aimed to clarify the detoxification effects of GA from Hezi on MA-induced cardiotoxicity and whether the detoxification mechanism is related to the TRPV1 channel. Cell viability was determined by methyl thiazol tetrazolium (MTT), and lactate dehydrogenase (LDH) leakage rate was determined by ELISA. Hoechst 33258, JC-1, DCFH-DA, and Fluo-3 AM staining were conducted to detect apoptosis, mitochondrial membrane potential, reactive oxygen species (ROS), and Ca2+ respectively; TRPV1 channel current was recorded by whole-cell patch-clamp technology to observe the effect of GA and MA alone or in combination on TRPV1 channel. The results showed that GA exhibited pronounced detoxification effects on MA-induced cardiotoxicity. GA significantly inhibited the MA-induced decrease in cell viability; suppressed the MA-induced LDH leakage rate, apoptosis, and the release of ROS and Ca2+; and alleviated the reduction of mitochondrial membrane potential. We found that MA-induced cardiotoxicity was significantly attenuated in H9c2 cells pretreated with the TRPV1 antagonist BCTC. In the whole-cell patch-clamp experiment, the TRPV1 channel current increase was caused by the GA and MA treatment, whereas it was reduced by the cotreatment of GA and MA. Our data demonstrate that GA in Hezi can reduce MA-induced cardiotoxicity by inhibiting intracellular Ca2+ influx, restoring mitochondrial membrane potential, and reducing apoptosis. The detoxification mechanism may be related to the desensitization of the TRPV1 channel by the combined application of MA and GA.

Integrated virtual screening and in vitro studies for exploring the mechanism of triterpenoids in Chebulae Fructus alleviating mesaconitine-induced cardiotoxicity via TRPV1 channel

“…We also demonstrated that chebulagic acid in CF has a protective effect on myocardial mitochondria, and can inhibit aconitine-induced Ca 2+ overload in myocardial cells, thereby reducing the occurrence of arrhythmia ( Han et al, 2020 ). In addition, we also found that the TRPV1 channel have an important role in exerting their attenuated effects, the gallic acid in CF may achieve the effect of detoxification by rapid desensitization of TRPV1 channel on the cells ( Han et al, 2022b ; Mi et al, 2023 ).…”

“…They reduce cardiotoxicity by involving numerous protein functions, signaling pathways, and biological processes ( Li et al, 2018 ). Our research group has verified that tannins and phenolic acids from CF can significantly reduce the mitochondrial membrane potential elevation and intracellular Ca 2+ overload caused by aconitine and MA in rat H9c2 cardiomyocytes through the TRPV1 channel, thereby improving mitochondrial function and inhibiting the cardiotoxicity of aconitine ( Han et al, 2020 ; Han et al, 2022b ; Han et al, 2022c ). When we were committed to the study of tannins, we accidentally found that triterpenoids in CF, while not as abundant as tannins, have a significant impact on antioxidant and cardioprotective properties.…”

“…The previous study found that the DDAs in aconitums caused intracellular calcium overload due to excessive activation of the TRPV1 channel in H9c2 cardiomyocytes. This led to damage to various organelles, resulting in a significant amount of apoptosis, which is one of the ways aconitum herbs produce cardiotoxicity ( Han et al, 2022b ; Han et al, 2022c ).…”

Deciphering the toxicity-effect relationship and action patterns of traditional Chinese medicines from a smart data perspective: a comprehensive review

“…In clinical applications, the combination of drugs can enhance the effect and mitigate aconite toxicity. Gallic acid can mitigate the toxicity of neoaconitine by inhibiting intracellular Ca 2+ influx, restoring mitochondrial membrane potential, and reducing apoptosis ( Han S. et al, 2022 ). In addition, berberine was found to prevent aconite-induced acute myocardial injury and arrhythmia, which may be related to the inhibition of aconitine-induced delayed depolarization and triggering activity ( Chen et al, 2018 ).…”