Ren (Armoracia rusticana, G. Gaertn, B. Mey. and Scherb.) je otporna višegodišnja biljka koja se uzgaja zbog specifičnog intenzivnog i ljutog ukusa. Cilj ovog rada bio je ispitivanje antitumorske i hepatoprotektivne aktivnosti ekstrakata i soka korena rena in vitro i in vivo. Primenom tečno-tečne ekstrakcije izolovane su i razdvojene polarne od nepolarnih komponenti, a HPLC metodom identifikovana su i kvantifikovana polifenolna jedinjenja u dobijenim ekstraktima i soku korena rena. Antiproliferativna aktivnost ekstrakata i soka korena rena ispitana je in vitro na tumorskim ćelijskim linijama: karcinoma grlića materice (HeLa), adenokarcinoma dojke (MCF7 i MDA-MB-231), adenokarcinoma debelog creva (HT-29), adenokarcinoma pluća (A549), adenokarcinoma prostate (PC-3), karcinoma kože (Hs 294T), karcinoma jetre (Hep G2), kao i na ćelijskim linijama karcinoma jetre pacova (H-4-II-E) i normalnim fetalnim ćelijskim linijama pluća (MPC-5) upotrebom Sulforodamin B testa. Mehanizam ćelijske smrti određen je detekcijom apoptoze i nekroze upotrebom Cell Death Detection ELISAPLUS kompleta. In vivo je ispitana antitumorska aktivnosti ekstrakata i soka korena rena na ćelijama Ehrlich-ovog ascitnog karcinoma (EAK) implantiranih NMRI miševima, kao i antioksidantna i hepatoprotektivna aktivnost kod intoksikacije jetre indukovane ugljen-tertrahloridom (CCL4) kod miševa. Ispitana je aktivnost osam ekstrakata korena rena i to iz pulpe: dihlormetanski (E1), hloroformski (E2), butanolni (E8) i vodeni (E7) i iz soka: dihlormetanski (E3), hloroformski (E4), butanolni (E6) i vodeni (E5) i sok korena rena (J9). Dihlormetanski ekstrakti korena rena imali su najveći sadržaj katehina, p-hidroksibenzoeve, siringinske i galne kiseline (pulpa, E1) i epikatehina (sok, E3). Hloroformski ekstrakt pulpe (E2) imao je visok sadržaj kvercetina i kemferola, dok je sok korena rena sadržao veće koncentracije katehina i galne kiseline. Rezultati su pokazali snažnu i neselektivnu antiproliferativnu aktivnost hloroformskih i dihlormetanskih ekstrakata i soka korena rena, sa najsnažnijim delovanjem na ćelijske linije jetre, dojke i pluća. Dobijene IC50 vrednosti bile su pri niskim koncentracijama (IC50 = 3,49-28,46 μg/mL) i visokim razblaženjima (IC50 = 418-1590). Sok (J9) i hloroformski ekstrakt soka rena (E4) pokazali su snažnu, nepoželjnu sposobnost indukcije nekroze. Ekstrakti i sok korena rena uticali su na povećanje oksidativnog stresa u ćelijama Ehrlich-ovog ascitnog karcinoma (EAK). Kod životinja koje su pretretirane sokom korena rena (J9) i posttretirane hloroformskim ekstraktom pulpe (E2) i dihlormetanskim ekstraktom soka (E3) postojalo je značajno povećanje aktivnosti superoksid dismutaze (SOD), ksantin oksidaze (XOD), glutation peroksidaze (GSHPx), glutation reduktaze (GR), intenziteta lipidne peroksidacije (LPx), kao i smanjenje aktivnosti katalaze (CAT) i nivoa glutationa (GSH) u ćelijama EAK. Efekti ekstrakata i soka korena rena na oksidativni stres kod hepatotoksičnog oštećenja jetre indukovanog ugljen-tetrahloridom bili su procenjeni merenjem parametara antioksidantne aktivnosti, kao i biohemijskih parametara funkcije jetre. Pretretmani sokom korena rena, hloroformskim ekstraktom pulpe (E2) i dihlormetanskim ekstraktom soka (E3), pre aplikacije CCl4, uticali su na značajno povećanje aktivnosti CAT, SOD, GR i nivoa GSH, kao i na značajno smanjenje aktivnosti XOD, GSHPx i intenziteta LPx, u odnosu na grupu životinja intoksikovanu CCl4-om. Takođe, pretretmani sa sokom korena rena, kao i hloroformskim i dihlormetanskim ekstraktima pre aplikacije CCl4 uticali su na značajno smanjenje aktivnosti ALT/AST i koncentracije hidroksiprolina, koje su bile povišene kod životinja nakon intoksikacije CCl4-om. Može se zaključiti da je detektovano snažno i neselektivno antiproliferativno dejstvo in vitro hloroformskih i dihlormetanskih ekstrakata i soka korena rena, sa nekrozom kao osnovnim mehanizmom indukovane ćelijske smrti. Rezultati dobijeni u in vivo ispitivanjima ukazali su da sok korena rena (J9), hloroformski ekstrakt pulpe (E2) i dihlormetanski ekstrakti soka (E3) ispoljavaju potencijalnu antitumorsku aktivnost prema ćelijama EAK, kao i potencijalnu antioksidantnu i hepatoprotektivnu aktivnost prevenirajući oštećenja jetre indukovana hepatotoksičnim CCl4-om., Horseradish (Armoracia rusticana, G. Gaertn, B. Mey. and Scherb.) is a hardy perennial herb, cultivated for its delicious, pungency and cooling taste. The aim of this study was to investigate antitumour and hepatoprptective activity of horseradish (Armoracia rusticana, Brasicaceae) root juice and extracts in vitro and in vivo. Liquid-liquid extraction of polar and non-polar compounds was used and polyphenolic compounds were identified and quantified by HPLC analysis. Antiproliferative activity was examinated in vitro on human cervix carcinoma (HeLa), breast adenocarcinoma (MCF7, MDA-MB-231), colon adenocarcinoma (HT-29), lung adenocarcinoma (A549), prostate adenocarcinoma (PC-3), melanocyte carcinoma (Hs 294T), hepatocyte carcinoma (Hep G2), as well as rat hepatocyte carcinoma (H-4-II-E), and normal human fetal lung (MRC-5) cell line using Sulforhodamine B assay. The mechanism of cell-death in cell line was determinated using Cell Death Detection ELISAPLUS kit. The antiproliferative activity of horseradish root juice and extracts against Ehrlich ascites carcinoma (EAC) in Hannover National Medical Institute (Hann:NMRI) mice, as well as antioxidant and hepatoprotective activity against carbon tetrachloride (CCl4)-induced liver injury in mice were examinated in vivo. This investigation was performed by use of dichloromethane (E1), chloroform (E2), butanol (E8) and aqueous (E7) extracts of horseradish pulp; dichloromethane (E3), chloroform (E4), butanol (E6) and aqueous (E5) extracts of horseradish juice, and unaltered horseradish juice (J9). Dichloromethane extracts had the highest content of catechin, p-hydroxybenzoic, syringic and gallic acid (pulp, E1), and epicatechin (juice, E3). Choroform pulp extract (E2) contained quercetin and kaempferol while gallic acid and catechin were mostly found in the juice (J9). The results showed strong and non-selective antiproliferative activity of chloroform and dichloromethane extracts and root juice - highest being towards liver, breast and lung tissue cells. IC50 values of extracts and juice were obtained in low range of concentrations (IC50 = 3,49-28,46 μg/mL) and high range of dilutions (IC50 = 418-1590). High and unfavorable potential of horseradish juice (J9) and chloroform juice extract (E4) to induce necrotic cell death was detected. Both the extracts and juice caused an increase of oxidative stress in EAC cells. Animals pre-treated with horseradish root juice (J9) and post-treated with chloroform pulp (E2) or dichloromethane juice (E3) extracts showed increased activities of superoxide dismutase (SOD), xanthine oxidase (XOD), glutathione peroxidase (GSHPx), glutathione reductase (GR) and the intensity of lipid peroxidation (LPx). On the other hand, catalase (CAT) activity and the amount of glutathione (GSH) significantly decreased in EAC cells. Effects of horseradish root juice and extracts on CCl4-induced oxidative stress were evaluated by measuring stress parameters and biochemical parameters of liver function. Pre-treatment with horseradish root juice, pulp chloroform (E2) and dichloromethane juice (E3) extracts, before application of CCl4, led to a significant increase in CAT, SOD, GR and GSH levels, and significantly decreased XOD, GSHPx and LPx levels, bringing them closer to the values of the EAC control group. Also, pre-treatment with horseradish root juice and chloroform and dichloromethane extracts, before application of CCl4 led to significant decrease in ALT/AST activities and hydroxyproline concentration, which were elevated in CCl4-intoxicated animals. It could be conclude that strong and non-selective in vitro antiproliferative activity of chloroform and dichloromethane extracts and root juice of horseradish was detected, with necrosis as a main mechanism of induced cell death. Obtained results suggest that the horseradish root juice (J9), as well as pulp chloroform (E2) and dichloromethane juice (E3) extracts exert antitumour effects on Ehrlich ascites carcinoma (EAC) and antioxidant and hepatoprotective effects against CCl4-induced liver toxicity in vivo.