Caffeic Acid Phenethyl Ester Protects Kidney Mitochondria against Ischemia/Reperfusion Induced Injury in an In Vivo Rat Model

Abstract

To improve ischemia/reperfusion tolerance, a lot of attention has been focused on natural antioxidants. Caffeic acid phenethyl ester (CAPE), an active component of the resinous exudates of the buds and young leaves of Populus nigra L., Baccharis sarothroides A., etc., and of propolis, possesses unique biological activities such as anti-inflammatory, antioxidant, immunomodulating, and cardioprotective effects, among others. There is a lack of studies showing a link between the antioxidant potential of CAPE and the mechanism of protective action of CAPE at the level of mitochondria, which produces the main energy for the basic functions of the cell. In the kidney, ischemia/reperfusion injury contributes to rapid kidney dysfunction and high mortality rates, and the search for biologically active protective compounds remains very actual. Therefore, the aim of this study was to identify the antioxidant potential of CAPE and to investigate whether CAPE can protect rat kidney mitochondria from in vivo kidney ischemia/reperfusion induced injury. We found that CAPE (1) possesses antioxidant activity (the reducing properties of CAPE are more pronounced than its antiradical properties); CAPE effectively reduces cytochrome c; (2) protects glutamate/malate oxidation and Complex I activity; (3) preserves the mitochondrial outer membrane from damage and from the release of cytochrome c; (4) inhibits reactive oxygen species (ROS) generation in the Complex II (SDH) F site; (5) diminishes ischemia/reperfusion-induced LDH release and protects from necrotic cell death; and (6) has no protective effects on succinate oxidation and on Complex II +III activity, but partially protects Complex II (SDH) from ischemia/reperfusion-induced damage. In summary, our study shows that caffeic acid phenethyl ester protects kidney mitochondrial oxidative phosphorylation and decreases ROS generation at Complex II in an in vivo ischemia/reperfusion model, and shows potential as a therapeutic agent for the development of pharmaceutical preparations against oxidative stress-related diseases.

Antioxidant and Mitochondria-Targeted Activity of Caffeoylquinic-Acid-Rich Fractions of Wormwood (Artemisia absinthium L.) and Silver Wormwood (Artemisia ludoviciana Nutt.)

“…Moreover, our in vitro results using the FRAP method revealed that both fractions from both Artemisia species' herb acetone extracts possess reducing properties. Therefore, we assume that Artemisia species fractions rich in caffeoylquinic acids can be suitable candidates for the modulation of mitochondrial function and regulation of mitochondrial redox state in the cells by suppressing ROS production and reducing cytochrome c. This finding is in agreement with our previous study showing that the ester of caffeic acid (caffeic acid phenethyl ester), which has a similar chemical structure to caffeoylquinic acids, also effectively reduces cytochrome c [28]. Our results are supported by the results of other scientists who have also investigated the role of cytochrome c redox state in the occurrence of apoptosis.…”

“…For antioxidant activity evaluation, all tested fractions and acetone extracts were dissolved in 70% ethanol until complete dissolution, obtaining a concentration of 0.5 mg/mL. Spectrophotometric assays were implemented as described by Raudone et al [ 27 ] and in our previous report [ 28 ].…”

“…Preparation of isolated kidney mitochondria of Wistar rats was performed using a differential centrifugation method, as described in [ 28 ].…”

“…The mitochondrial functions were measured using an Oxygraph-2k high-resolution respirometry system (OROBOROS Instruments, Innsbruck, Austria) in the incubation medium, as described in [ 28 ]. The routine mitochondrial respiration (V 0 ) was recorded in the medium supplemented with mitochondria and 5 mM glutamate + 5 mM malate as respiratory substrates.…”

“…The reduction of cytochrome c was recorded spectrophotometrically using a Helios Gama spectrophotometer as described in our previous study [ 28 ]. Briefly, the absorption spectra at 500–600 nm of various fractions in 1 mL of incubation medium were measured, and cytochrome c (20 µM) was added to the same buffer.…”