Cytoprotective properties of the ethyl acetate extracts from Anacyclus maroccanus Ball. e Anacyclus radiatus Loisel aerial parts

Anacyclus species (Fam. Asteraceae) are endemic plants from Morocco greatly exploited by the local population for primary health care, as inexpensive and available sources of drugs. They are traditionally used by Moroccan people to treat several ailments, among which digestive disorders, likely due to their antioxidant, anti-inflammatory, analgesic, and antimicrobial properties [1]. In our previous study we highlighted promising antioxidant, hypoglycemic, and antiglycative properties of the ethyl acetate extracts from the aerial parts of two Anacyclus ecotypes, namely A. maroccanus Ball. and A. radiatus Loisel, likely associated to their polyphenolic composition [1]. In line with this evidence, present study was aimed at evaluating the cytoprotective properties of the ethyl acetate extracts from A. maroccanus and A. radiatus towards the oxidative damage induced by the pro-oxidant agent tert-butyl hydroperoxide (tBOOH, 500 μM) and hyperglycemia (6 mg/ml glucose) in human cell lines [2]. To this aim, pancreatic adenocarcinoma (Bx-PC3) and cholangiocarcinoma (Mz-ChA-1) cells, and noncancerous intrahepatic cholangiocytes (H69) were exploited. The cells were exposed to the treatments for 24 h under both pre-treatment and co-treatment protocols, then the the cytotoxicity was evaluated by the MTT assay and the intracellular ROS levels by the 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) test, according to Di Giacomo et al. [3]. Preliminary cytotoxicity assays highlighted the extracts were nontoxic up to 100 μg/ml in all the tested cells. Under our experimental conditions, tBOOH induced about a 50-60% reduction of cell viability in Mz-ChA-1, Bx-PC3, and H69 cells. As expected, the extracts of both ecotypes were able to counteract the tBOOH cytotoxicity up to 70-80% in Mz-ChA-1. The extracts also decreased the intracellular ROS levels, almost doubled by tBOOH, with a greater efficacy of A. radiatus in Bx-PC3 with respect to A. maroccanus (60% vs 40% increase of cell viability respectively); both samples reduced the ROS levels by about 52% and 65%, respectively. At last, A. maroccanus completely restored cell viability and lowered the ROS basal levels, despite a weak effect of A. radiatus in H69 cells. Regarding the hyperglycemia model, glucose (6 mg/ml) induced a 45% and 60% lowering of cell viability in Bx-PC3 and H69 cells. The A. maroccanus and A. radiatus extracts induced only a slight cytoprotection, in terms of cell viability and ROS levels in H69 cells. By contrast, they showed protective properties against 1° Congresso intersocietà sui prodotti vegetali per la salute: Il ruolo delle piante medicinali nella medicina moderna PADOVA, 15-17 Giugno 2023 Dipartimento di Scienze del Farmaco, Via Marzolo, 5 hyperglycemia damage in H69 cells, by inducing a 24% increase of cell viability and a 32% lowering ROS. Altogether, present results highlight a possible interest in these species as natural sources of bioactive compounds and/or phytocomplexes and suggest further studies in order to characterize their possible application for nutraceutical purposes, with possible benefits for Moroccan economy. References 1. Sissi et al., Molecules 2022;27(3):692. 2. Di Sotto et al., Oxid Med Cell Longev. 2018;2018:5919237. 3. Di Giacomo et al., Pharmaceuticals 15(8), 987, 2022.