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Dominique Pessayre - One of the best experts on this subject based on the ideXlab platform.
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human epoxide hydrolase is the target of Germander autoantibodies on the surface of human hepatocytes enzymatic implications
Advances in Experimental Medicine and Biology, 2001Co-Authors: Jacqueline Loeper, Dominique Pessayre, Veronique De Berardinis, Claude Moulis, Philippe Beaune, Denis PomponAbstract:Western countries observed an increased interest for the use of herbal medicines because of their supposed safety in contrast to chemical drugs. Wild Germander (Teuchrium chamaedrys L) was traditionally used as a folk medicine for its for and antiseptic properties. In 1991, Germander consumed to treat obesity, caused an epidemic of cytolytic hepatitis. Thirty cases of hepatotoxicity were first reported including cases with positive rechallengge. For these patients, an early recurrence was observed despite lack of other features of hypersensitivity (Castot and Larrey, 1992). In mice, Germander toxicity required CYP3A-dependent metabolism (Loeper et al 1994). More specifically, the metabolic activation of the furan ring of the diterpenoid teucrin A (TA) (Kouzi et al 1994). TA- toxicity was via CYP3A-generated electrophilic metabolites that were detoxified by glutathione conjugation, depleted cellular thiols and caused apoptosis in isolated rat hepatocytes atocytes (Lekehal et al 1996; Fau et al 1997). However, these death processes did not explain the immunologial cal features observed in several cases of Germander-induced hepatitis. To explain this immune process, patient’s sera consuming Germander tea in great quantity were Tested by Western blot analysis. They contained autoantibodies directed against human microsomal epoxide hydrolase (hmEH), that was located both, in the endoplasmic reticulum and the plasma membrane (PM) of human hepatocyte and hmEH- expressing yeast. Germander-induced autoantibodies (GIAA) were shown to recognize hmEH on the cell surface. To implicate hmEH in the metabolic activation of TA we used a “humanized” yeast strain expressing human P450-reductase and cytochrome b5 transformed with CYP3A4 and/or hmEH cDNAs. TA was metabolized by CYP3A4 into a and hmEH with TA inactivated hmEH in a time-dependent-manner, in agreement with formation of reactive teuchrin A-metabolite that could covalently alter and inhibit hmEH. This modified enzyme may bypass the immunologic tolerance that normally exists for hmEH.
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human microsomal epoxide hydrolase is the target of Germander induced autoantibodies on the surface of human hepatocytes
Molecular Pharmacology, 2000Co-Authors: Veronique De Berardinis, Dominique Pessayre, Claude Moulis, Philippe Beaune, Denis Pompon, Michele Maurice, Jacqueline LoeperAbstract:Germander, a plant used in folk medicine, caused an epidemic of cytolytic hepatitis in France. In about half of these patients, a rechallenge caused early recurrence, suggesting an immunoallergic type of hepatitis. Teucrin A (TA) was found responsible for the hepatotoxicity via metabolic activation by CYP3A. In this study, we describe the presence of anti-microsomal epoxide hydrolase (EH) autoantibodies in the sera of patients who drank Germander teas for a long period of time. By Western blotting and immunocytochemistry, human microsomal EH was shown to be present in purified plasma membranes of both human hepatocytes and transformed spheroplasts and to be exposed on the cell surface where affinity-purified Germander autoantibodies recognized it as their autoantigen. Immunoprecipitation of EH activity by Germander-induced autoantibodies confirmed this finding. These autoantibodies were not immunoinhibitory. The plasma membrane-located EH was catalytically competent and may act as target for reactive metabolites from TA. To test this hypothesis CYP3A4 and EH were expressed with human cytochrome P450 reductase and cytochrome b(5) in a "humanized" yeast strain. In the absence of EH only one metabolite was formed. In the presence of EH, two additional metabolites were formed, and a time-dependent inactivation of EH was detected, suggesting that a reactive oxide derived from TA could alkylate the enzyme and trigger an immune response. Antibodies were found to recognize TA-alkylated EH. Recognition of EH present at the surface of human hepatocytes could suggest an (auto)antibody participation in an immune cell destruction.
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diterpenoids from Germander an herbal medicine induce apoptosis in isolated rat hepatocytes
Gastroenterology, 1997Co-Authors: Daniel Fau, Mounia Lekehal, G Farrell, Amelie Moreau, C Moulis, Gerard Feldmann, D Haouzi, Dominique PessayreAbstract:Abstract BACKGROUND & AIMS: Germander was withdrawn from the market after its use for weight control caused an epidemic of hepatitis. Its toxicity was shown to be caused by diterpenoids and their cytochrome P4503A- mediated metabolic activation into electrophilic metabolites that deplete cellular thiols. The aim of the present study was to determine the mechanisms of cell death. METHODS: Isolated rat hepatocytes were incubated for 2 hours with Germander diterpenoids (100 micrograms/mL). RESULTS: Diterpenoids decreased cell glutathione, increased cytosolic [Ca2+], activated Ca(2+)-dependent tissue transglutaminase forming a cross-linked protein scaffold, and caused internucleosomal DNA fragmentation and the ultrastructural features of apoptosis. Cell death was prevented by decreasing metabolic activation (with troleandomycin), preventing depletion of glutathione (with cystine), blocking activation of Ca(2+)-modulated enzymes (with calmidazolium), or inhibiting internucleosomal DNA fragmentation (with aurintricarboxylic acid). Apoptosis was increased and diterpenoids caused overexpression of p53 and interleukin 1 beta-converting enzyme in rats treated with dexamethasone (cytochrome P4503A inducer). Apoptosis was also increased by a diet deficient in sulfur amino acids. CONCLUSIONS: The Germander furano diterpenoids cause apoptosis within 2 hours in isolated rat hepatocytes. Electrophilic metabolites may stimulate apoptosis by decreasing thiols, increasing [Ca2+], and activating Ca(2+)-dependent transglutaminase and endonucleases. (Gastroenterology 1997 Oct;113(4):1334-46)
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hepatotoxicity of the herbal medicine Germander metabolic activation of its furano diterpenoids by cytochrome p450 3a depletes cytoskeleton associated protein thiols and forms plasma membrane blebs in rat hepatocytes
Hepatology, 1996Co-Authors: Mounia Lekehal, Claire Moulis, J M Lereau, Dominique Pessayre, Isabelle FourastéAbstract:Abstract Several herbal remedies have produced hepatitis in humans. The medicinal plant, Germander, was recalled after its use as an adjuvant to slimming diets resulted in an epidemic of hepatitis in France. We studied the hepatotoxicity of Germander in isolated rat hepatocytes. A crude fraction containing the diverse furano diterpenoids of Germander, or the purified main constituents of this fraction, teucrin A and teuchamaedryn A, were hepatotoxic (correction for hepatototoxic), but not fractions containing more polar or lipophilic constituents. [ 3 H]Teucrin A covalently bound to hepatocyte proteins. The furano diterpenoid fraction decreased cell glutathione and cytoskeleton- associated protein thiols, and led to formation of plasma membrane blebs and cell demise. Pretreatment of male rats with troleandomycin, an inhibitor of cytochrome P450 3A (CYP3A), slowed the depletion of glutathione and decreased toxicity, whereas dexamethasone, an inducer of CYP3A, had opposite effects. Female rat hepatocytes, which poorly express CYP3A, exhibited little toxicity, unless the animals were treated with dexamethasone. Feeding male rats with a sulfur amino acid- deficient diet decreased cell glutathione and enhanced toxicity, whereas supplementation of the standard diet with cystine had opposite effects. We conclude that the furano diterpenoids of Germander are activated by CYP3A into electrophilic metabolites that deplete glutathione and cytoskeleton-associated protein thiols and form plasma membrane blebs. We suggest that studies in isolated hepatocytes be included in the preclinical assessment of herbal remedies. (Hepatology 1996 Jul;24(1):212-8)
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hepatotoxicity of the herbal medicine Germander metabolic activation of its furano diterpenoids by cytochrome p450 3a depletes cytoskeleton associated protein thiols and forms plasma membrane blebs in rat hepatocytes
Hepatology, 1996Co-Authors: Mounia Lekehal, Claire Moulis, Isabelle Fourasté, J M Lereau, Dominique Pessayre, Daniel FauAbstract:Several herbal remedies have produced hepatitis in humans. The medicinal plant, Germander, was recalled after its use as an adjuvant to slimming diets resulted in an epidemic of hepatitis in France. We studied the hepatotoxicity of Germander in isolated rat hepatocytes. A crude fraction containing the diverse furano diterpenoids of Germander, or the purified main constituents of this fraction, teucrin A and teuchamaedryn A, were hepatotoxic (correction for hepatototoxic), but not fractions containing more polar or lipophilic constituents. [3H]Teucrin A covalently bound to hepatocyte proteins. The furano diterpenoid fraction decreased cell glutathione and cytoskeleton-associated protein thiols, and led to formation of plasma membrane blebs and cell demise. Pretreatment of male rats with troleandomycin, an inhibitor of cytochrome P450 3A (CYP3A), slowed the depletion of glutathione and decreased toxicity, whereas dexamethasone, an inducer of CYP3A, had opposite effects. Female rat hepatocytes, which poorly express CYP3A, exhibited little toxicity, unless the animals were treated with dexamethasone. Feeding male rats with a sulfur amino acid-deficient diet decreased cell glutathione and enhanced toxicity, whereas supplementation of the standard diet with cystine had opposite effects. We conclude that the furano diterpenoids of Germander are activated by CYP3A into electrophilic metabolites that deplete glutathione and cytoskeleton-associated protein thiols and form plasma membrane blebs. We suggest that studies in isolated hepatocytes be included in the preclinical assessment of herbal remedies.
Daniel Fau - One of the best experts on this subject based on the ideXlab platform.
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diterpenoids from Germander an herbal medicine induce apoptosis in isolated rat hepatocytes
Gastroenterology, 1997Co-Authors: Daniel Fau, Mounia Lekehal, G Farrell, Amelie Moreau, C Moulis, Gerard Feldmann, D Haouzi, Dominique PessayreAbstract:Abstract BACKGROUND & AIMS: Germander was withdrawn from the market after its use for weight control caused an epidemic of hepatitis. Its toxicity was shown to be caused by diterpenoids and their cytochrome P4503A- mediated metabolic activation into electrophilic metabolites that deplete cellular thiols. The aim of the present study was to determine the mechanisms of cell death. METHODS: Isolated rat hepatocytes were incubated for 2 hours with Germander diterpenoids (100 micrograms/mL). RESULTS: Diterpenoids decreased cell glutathione, increased cytosolic [Ca2+], activated Ca(2+)-dependent tissue transglutaminase forming a cross-linked protein scaffold, and caused internucleosomal DNA fragmentation and the ultrastructural features of apoptosis. Cell death was prevented by decreasing metabolic activation (with troleandomycin), preventing depletion of glutathione (with cystine), blocking activation of Ca(2+)-modulated enzymes (with calmidazolium), or inhibiting internucleosomal DNA fragmentation (with aurintricarboxylic acid). Apoptosis was increased and diterpenoids caused overexpression of p53 and interleukin 1 beta-converting enzyme in rats treated with dexamethasone (cytochrome P4503A inducer). Apoptosis was also increased by a diet deficient in sulfur amino acids. CONCLUSIONS: The Germander furano diterpenoids cause apoptosis within 2 hours in isolated rat hepatocytes. Electrophilic metabolites may stimulate apoptosis by decreasing thiols, increasing [Ca2+], and activating Ca(2+)-dependent transglutaminase and endonucleases. (Gastroenterology 1997 Oct;113(4):1334-46)
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hepatotoxicity of the herbal medicine Germander metabolic activation of its furano diterpenoids by cytochrome p450 3a depletes cytoskeleton associated protein thiols and forms plasma membrane blebs in rat hepatocytes
Hepatology, 1996Co-Authors: Mounia Lekehal, Claire Moulis, Isabelle Fourasté, J M Lereau, Dominique Pessayre, Daniel FauAbstract:Several herbal remedies have produced hepatitis in humans. The medicinal plant, Germander, was recalled after its use as an adjuvant to slimming diets resulted in an epidemic of hepatitis in France. We studied the hepatotoxicity of Germander in isolated rat hepatocytes. A crude fraction containing the diverse furano diterpenoids of Germander, or the purified main constituents of this fraction, teucrin A and teuchamaedryn A, were hepatotoxic (correction for hepatototoxic), but not fractions containing more polar or lipophilic constituents. [3H]Teucrin A covalently bound to hepatocyte proteins. The furano diterpenoid fraction decreased cell glutathione and cytoskeleton-associated protein thiols, and led to formation of plasma membrane blebs and cell demise. Pretreatment of male rats with troleandomycin, an inhibitor of cytochrome P450 3A (CYP3A), slowed the depletion of glutathione and decreased toxicity, whereas dexamethasone, an inducer of CYP3A, had opposite effects. Female rat hepatocytes, which poorly express CYP3A, exhibited little toxicity, unless the animals were treated with dexamethasone. Feeding male rats with a sulfur amino acid-deficient diet decreased cell glutathione and enhanced toxicity, whereas supplementation of the standard diet with cystine had opposite effects. We conclude that the furano diterpenoids of Germander are activated by CYP3A into electrophilic metabolites that deplete glutathione and cytoskeleton-associated protein thiols and form plasma membrane blebs. We suggest that studies in isolated hepatocytes be included in the preclinical assessment of herbal remedies.
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hepatotoxicity of Germander in mice
Gastroenterology, 1994Co-Authors: Jacqueline Loeper, Claude Moulis, Veronique Descatoire, Philippe Letteron, Claude Degott, Patrick M Dansette, Daniel Fau, Dominique PessayreAbstract:Abstract Background/Aims: An epidemic of hepatitis due to Germander teas or capsules recently occurred in France. The aim of the present study was to show the hepatotoxicity of Germander and determine its mechanism in mice. Methods: A Germander tea lyophilisate and a fraction that isolated and concentrated 10-fold the furano neo -clerodane diterpenoids of the lyophilisate were prepared. Results: (1) Intragastric administration of the lyophilisate (1.25 g/kg) or the furano neo -clerodane diterpenoid fraction (0.125 mg/kg) produced similar midzonal liver cell necrosis at 24 hours in mice. (2) Toxicity was prevented by pretreatment with a single dose of troleandomycin (a specific inhibitor of cytochromes P4503A) and enhanced by pretreatment with dexamethasone or clotrimazole (two inducers of cytochromes P4503A). (3) Toxicity was attenuated by pretreatment with butylated hydroxyanisole or clofibrate (two inducers of microsomal epoxide hydrolase) and markedly increased by phorone-induced glutathione depletion. Conclusions: We conclude that Germander constituents (probably its furano neo -clerodane diterpenoids) are transformed by cytochromes P450 (particularly P4503A) into hepatotoxic metabolites. The metabolites (probably epoxides) are partly inactivated by glutathione and probably epoxide hydrolase.
Mounia Lekehal - One of the best experts on this subject based on the ideXlab platform.
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diterpenoids from Germander an herbal medicine induce apoptosis in isolated rat hepatocytes
Gastroenterology, 1997Co-Authors: Daniel Fau, Mounia Lekehal, G Farrell, Amelie Moreau, C Moulis, Gerard Feldmann, D Haouzi, Dominique PessayreAbstract:Abstract BACKGROUND & AIMS: Germander was withdrawn from the market after its use for weight control caused an epidemic of hepatitis. Its toxicity was shown to be caused by diterpenoids and their cytochrome P4503A- mediated metabolic activation into electrophilic metabolites that deplete cellular thiols. The aim of the present study was to determine the mechanisms of cell death. METHODS: Isolated rat hepatocytes were incubated for 2 hours with Germander diterpenoids (100 micrograms/mL). RESULTS: Diterpenoids decreased cell glutathione, increased cytosolic [Ca2+], activated Ca(2+)-dependent tissue transglutaminase forming a cross-linked protein scaffold, and caused internucleosomal DNA fragmentation and the ultrastructural features of apoptosis. Cell death was prevented by decreasing metabolic activation (with troleandomycin), preventing depletion of glutathione (with cystine), blocking activation of Ca(2+)-modulated enzymes (with calmidazolium), or inhibiting internucleosomal DNA fragmentation (with aurintricarboxylic acid). Apoptosis was increased and diterpenoids caused overexpression of p53 and interleukin 1 beta-converting enzyme in rats treated with dexamethasone (cytochrome P4503A inducer). Apoptosis was also increased by a diet deficient in sulfur amino acids. CONCLUSIONS: The Germander furano diterpenoids cause apoptosis within 2 hours in isolated rat hepatocytes. Electrophilic metabolites may stimulate apoptosis by decreasing thiols, increasing [Ca2+], and activating Ca(2+)-dependent transglutaminase and endonucleases. (Gastroenterology 1997 Oct;113(4):1334-46)
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hepatotoxicity of the herbal medicine Germander metabolic activation of its furano diterpenoids by cytochrome p450 3a depletes cytoskeleton associated protein thiols and forms plasma membrane blebs in rat hepatocytes
Hepatology, 1996Co-Authors: Mounia Lekehal, Claire Moulis, J M Lereau, Dominique Pessayre, Isabelle FourastéAbstract:Abstract Several herbal remedies have produced hepatitis in humans. The medicinal plant, Germander, was recalled after its use as an adjuvant to slimming diets resulted in an epidemic of hepatitis in France. We studied the hepatotoxicity of Germander in isolated rat hepatocytes. A crude fraction containing the diverse furano diterpenoids of Germander, or the purified main constituents of this fraction, teucrin A and teuchamaedryn A, were hepatotoxic (correction for hepatototoxic), but not fractions containing more polar or lipophilic constituents. [ 3 H]Teucrin A covalently bound to hepatocyte proteins. The furano diterpenoid fraction decreased cell glutathione and cytoskeleton- associated protein thiols, and led to formation of plasma membrane blebs and cell demise. Pretreatment of male rats with troleandomycin, an inhibitor of cytochrome P450 3A (CYP3A), slowed the depletion of glutathione and decreased toxicity, whereas dexamethasone, an inducer of CYP3A, had opposite effects. Female rat hepatocytes, which poorly express CYP3A, exhibited little toxicity, unless the animals were treated with dexamethasone. Feeding male rats with a sulfur amino acid- deficient diet decreased cell glutathione and enhanced toxicity, whereas supplementation of the standard diet with cystine had opposite effects. We conclude that the furano diterpenoids of Germander are activated by CYP3A into electrophilic metabolites that deplete glutathione and cytoskeleton-associated protein thiols and form plasma membrane blebs. We suggest that studies in isolated hepatocytes be included in the preclinical assessment of herbal remedies. (Hepatology 1996 Jul;24(1):212-8)
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hepatotoxicity of the herbal medicine Germander metabolic activation of its furano diterpenoids by cytochrome p450 3a depletes cytoskeleton associated protein thiols and forms plasma membrane blebs in rat hepatocytes
Hepatology, 1996Co-Authors: Mounia Lekehal, Claire Moulis, Isabelle Fourasté, J M Lereau, Dominique Pessayre, Daniel FauAbstract:Several herbal remedies have produced hepatitis in humans. The medicinal plant, Germander, was recalled after its use as an adjuvant to slimming diets resulted in an epidemic of hepatitis in France. We studied the hepatotoxicity of Germander in isolated rat hepatocytes. A crude fraction containing the diverse furano diterpenoids of Germander, or the purified main constituents of this fraction, teucrin A and teuchamaedryn A, were hepatotoxic (correction for hepatototoxic), but not fractions containing more polar or lipophilic constituents. [3H]Teucrin A covalently bound to hepatocyte proteins. The furano diterpenoid fraction decreased cell glutathione and cytoskeleton-associated protein thiols, and led to formation of plasma membrane blebs and cell demise. Pretreatment of male rats with troleandomycin, an inhibitor of cytochrome P450 3A (CYP3A), slowed the depletion of glutathione and decreased toxicity, whereas dexamethasone, an inducer of CYP3A, had opposite effects. Female rat hepatocytes, which poorly express CYP3A, exhibited little toxicity, unless the animals were treated with dexamethasone. Feeding male rats with a sulfur amino acid-deficient diet decreased cell glutathione and enhanced toxicity, whereas supplementation of the standard diet with cystine had opposite effects. We conclude that the furano diterpenoids of Germander are activated by CYP3A into electrophilic metabolites that deplete glutathione and cytoskeleton-associated protein thiols and form plasma membrane blebs. We suggest that studies in isolated hepatocytes be included in the preclinical assessment of herbal remedies.
Natasa Nastic - One of the best experts on this subject based on the ideXlab platform.
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subcritical water extraction of antioxidants from mountain Germander teucrium montanum l
Journal of Supercritical Fluids, 2018Co-Authors: Natasa Nastic, Jaroslava Svarcgajic, Cristina Deleruematos, Simone Morais, Fatima M Barroso, Manuela M MoreiraAbstract:Abstract In the present work, antioxidant compounds from Teucrium montanum were extracted by subcritical water. The influence of extraction temperature and pressure on antioxidant activity of extracts has been investigated in terms of extraction yield (EY), total phenolic content (TPC), and DPPH-radical scavenging activity (DPPH-RSA) and ferric reducing antioxidant power (FRAP). Additionally, the compounds responsible for the antioxidant activity were identified and quantified by high performance liquid chromatography (HPLC). The highest EY (42.63%), TPC (174.61 ± 4.09 mg GAE/g DE) and antioxidant activity by DPPH-RSA (176.23 ± 8.76 mg TE/g DE) and FRAP (141.71 ± 5.21 mg AAE/g DE) were seen in extracts obtained at temperature of 160 °C and pressure of 10 bar. HPLC analysis revealed that naringin and gallic acid were the principle antioxidant compounds in subcritical extracts. According to the results, SWE has a great potential in exploitation of natural sources of bioactive compounds and production of pharmacologically-active fractions.
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subcritical water extraction as an environmentally friendly technique to recover bioactive compounds from traditional serbian medicinal plants
Industrial Crops and Products, 2018Co-Authors: Natasa Nastic, Jaroslava Svarcgajic, Cristina Deleruematos, Simone Morais, Fatima M Barroso, Manuela M Moreira, Cristina Soares, Pavle Maskovic, Visnja Gaurina Srcek, Igor SlivacAbstract:Abstract Subcritical water extraction (SWE) has become a popular green extraction technique for the isolation of different classes of compounds from natural matrices. Low price, safety and green character of water, good yields of target compounds and reduced energy consumption, make this technique favorable for potential industrial applications. The purpose of this study was to evaluate antioxidant, antimicrobial and cytotoxic activity of four medicinal plants traditionally used in folk medicine of Serbia. Black mulberry (Morus nigra L.), wall Germander (Teucrium chamaedrys L.), wild geranium (Geranium macrorrhizum L.) and comfrey (Symphytum officinale L.) were extracted by subcritical water at different temperatures. Antioxidant activity of the extracts was defined by conventional spectrophotometric methods, such as the total phenolic content (TPC), DPPH-radical scavenging activity (DPPH-RSA), ferric reducing antioxidant power (FRAP) and total antioxidant capacity (TAC) assessed by a DNA-based sensor. Additionally, the main phenolic compounds contributing to the antioxidant activity of the produced extracts were also identified and quantified by high performance liquid chromatography with diode array detection (HPLC-DAD). Antimicrobial properties of extracts were evaluated against eight microbial strains. Furthermore, the cytotoxic activity was observed for two human cancer cell lines and a cell line derived from murine fibroblast.
Manuela M Moreira - One of the best experts on this subject based on the ideXlab platform.
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subcritical water extraction of antioxidants from mountain Germander teucrium montanum l
Journal of Supercritical Fluids, 2018Co-Authors: Natasa Nastic, Jaroslava Svarcgajic, Cristina Deleruematos, Simone Morais, Fatima M Barroso, Manuela M MoreiraAbstract:Abstract In the present work, antioxidant compounds from Teucrium montanum were extracted by subcritical water. The influence of extraction temperature and pressure on antioxidant activity of extracts has been investigated in terms of extraction yield (EY), total phenolic content (TPC), and DPPH-radical scavenging activity (DPPH-RSA) and ferric reducing antioxidant power (FRAP). Additionally, the compounds responsible for the antioxidant activity were identified and quantified by high performance liquid chromatography (HPLC). The highest EY (42.63%), TPC (174.61 ± 4.09 mg GAE/g DE) and antioxidant activity by DPPH-RSA (176.23 ± 8.76 mg TE/g DE) and FRAP (141.71 ± 5.21 mg AAE/g DE) were seen in extracts obtained at temperature of 160 °C and pressure of 10 bar. HPLC analysis revealed that naringin and gallic acid were the principle antioxidant compounds in subcritical extracts. According to the results, SWE has a great potential in exploitation of natural sources of bioactive compounds and production of pharmacologically-active fractions.
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subcritical water extraction as an environmentally friendly technique to recover bioactive compounds from traditional serbian medicinal plants
Industrial Crops and Products, 2018Co-Authors: Natasa Nastic, Jaroslava Svarcgajic, Cristina Deleruematos, Simone Morais, Fatima M Barroso, Manuela M Moreira, Cristina Soares, Pavle Maskovic, Visnja Gaurina Srcek, Igor SlivacAbstract:Abstract Subcritical water extraction (SWE) has become a popular green extraction technique for the isolation of different classes of compounds from natural matrices. Low price, safety and green character of water, good yields of target compounds and reduced energy consumption, make this technique favorable for potential industrial applications. The purpose of this study was to evaluate antioxidant, antimicrobial and cytotoxic activity of four medicinal plants traditionally used in folk medicine of Serbia. Black mulberry (Morus nigra L.), wall Germander (Teucrium chamaedrys L.), wild geranium (Geranium macrorrhizum L.) and comfrey (Symphytum officinale L.) were extracted by subcritical water at different temperatures. Antioxidant activity of the extracts was defined by conventional spectrophotometric methods, such as the total phenolic content (TPC), DPPH-radical scavenging activity (DPPH-RSA), ferric reducing antioxidant power (FRAP) and total antioxidant capacity (TAC) assessed by a DNA-based sensor. Additionally, the main phenolic compounds contributing to the antioxidant activity of the produced extracts were also identified and quantified by high performance liquid chromatography with diode array detection (HPLC-DAD). Antimicrobial properties of extracts were evaluated against eight microbial strains. Furthermore, the cytotoxic activity was observed for two human cancer cell lines and a cell line derived from murine fibroblast.