Chlorogenic Acids, Acting via Calcineurin, Are the Main Compounds in Centella asiatica Extracts That Mediate Resilience to Chronic Stress in Drosophila melanogaster

Abstract

Common symptoms of depressive disorders include anhedonia, sleep problems, and reduced physical activity. Drugs used to treat depression mostly aim to increase serotonin signaling but these can have unwanted side effects. Depression has also been treated by traditional medicine using plants like Centella asiatica (CA) and this has been found to be well tolerated. However, very few controlled studies have addressed CA’s protective role in depression, nor have the active compounds or mechanisms that mediate this function been identified. To address this issue, we used Drosophila melanogaster to investigate whether CA can improve depression-associated symptoms like anhedonia and decreased climbing activity. We found that a water extract of CA provides resilience to stress induced phenotypes and that this effect is primarily due to mono-caffeoylquinic acids found in CA. Furthermore, we describe that the protective function of CA is due to a synergy between chlorogenic acid and one of its isomers also present in CA. However, increasing the concentration of chlorogenic acid can overcome the requirement for the second isomer. Lastly, we found that chlorogenic acid acts via calcineurin, a multifunctional phosphatase that can regulate synaptic transmission and plasticity and is also involved in neuronal maintenance.

Centella asiatica and its caffeoylquinic acid and triterpene constituents increase dendritic arborization of mouse primary hippocampal neurons and improve age-related locomotion deficits in Drosophila

“…First, we tested mixes of CQAs and TTs containing the compounds in concentrations equivalent to CAW (10 mg/mL). We previously showed that the levels of these compounds in CAW are not affected by the preparation of the food and are stable for the 7 days the flies are kept on a food vial ( Holvoet et al, 2023a ). The compounds and compound mixes were prepared in ethanol as 10x stock solutions and therefore the control flies also obtained 10% ethanol in yeast paste.…”

“…The preparation of the CA water extract used in this study (BEN-CAW-7) has been previously described in detail ( Holvoet et al, 2023a ; Yang et al, 2023a ). BEN-CA-6 plant material (4 kg) was extracted by boiling in deionized water (50 L) for 90 min.…”

“…Four purified triterpenes (TTs) and eight caffeoylquinic acids (CQAs) known to occur in CAW ( RdS et al, 2022 ) were purchased from commercial sources and their identity verified by us as previously described ( Holvoet et al, 2023b ). The content of these compounds in the CAW batch used in this study was determined by liquid chromatography coupled to multiple reaction monitoring mass spectrometry (LC-MRM-MS) as previously reported by us ( Holvoet et al, 2023b ; Yang et al, 2023b ).…”

“…Four purified triterpenes (TTs) and eight caffeoylquinic acids (CQAs) known to occur in CAW ( RdS et al, 2022 ) were purchased from commercial sources and their identity verified by us as previously described ( Holvoet et al, 2023b ). The content of these compounds in the CAW batch used in this study was determined by liquid chromatography coupled to multiple reaction monitoring mass spectrometry (LC-MRM-MS) as previously reported by us ( Holvoet et al, 2023b ; Yang et al, 2023b ). Content (% w/w) of the selected TT and CQA compounds in CAW were asiatic acid, 0.042%; madecassic acid, 0.075%; asiaticoside, 1.475%; madecassoside 3.589%; chlorogenic acid (3-CQA), 0.750%, cryptochlorogenic acid (4-CQA), 0.298%; neochlorogenic acid (5 CQA), 0.339%; 1,3-diCQA, 0.258%; 1,5- diCQA, 0.389%; Isochlorogenic acid A (3,5- diCQA), 0.177%; Isochlorogenic acid B (3,4- diCQA), 0.229%, and Isochlorogenic acid C (4,5- diCQA), 0.195%.…”

“…Application by yeast paste was also used when testing individual compounds and compound mixes. If not stated otherwise, the concentration of compounds was equivalent to their levels found in CAW as previously determined ( Yang et al, 2023a ; Holvoet et al, 2023b ). For treatment of Drosophila a 10x concentrated stock solution of the compound or compounds was prepared in ethanol and diluted 1:10 in yeast paste.…”