C Caliceti1, P Rizzo2, R Ferrari3, F Fortini4, G Aquila4, E Leoncini5, L Zambonin6, B Rizzo5, D Calabria7, P Simoni8, M Mirasoli9, M Guardigli10, S Hrelia5, A Roda9, A F G Cicero8. 1. Department of Chemistry "Giacomo Ciamician" - Alma Mater Studiorum, University of Bologna, Bologna, Italy; Centro Interdipartimentale di Ricerca Industriale Energia e Ambiente (CIRI EA) - Alma Mater Studiorum, University of Bologna, Bologna, Italy; Istituto Nazionale Biostrutture e Biosistemi (INBB), Roma, Italy. Electronic address: cristiana.caliceti@unibo.it. 2. Department of Morphology, Surgery and Experimental Medicine, University of Ferrara, Ferrara, Italy; Maria Cecilia Hospital, GVM Care&Research, E.S: Health Science Foundation, Cotignola, Italy. 3. Department of Medical Sciences, Cardiology and Laboratory for Technologies of Advanced Therapies (LTTA), University of Ferrara, Ferrara, Italy; Maria Cecilia Hospital, GVM Care&Research, E.S: Health Science Foundation, Cotignola, Italy. 4. Department of Medical Sciences, University of Ferrara, Ferrara, Italy. 5. Department for Life Quality Studies - Alma Mater Studiorum, University of Bologna, Bologna, Italy. 6. Department of Pharmacy and Biotechnology - Alma Mater Studiorum, University of Bologna, Bologna, Italy. 7. Centro Interdipartimentale di Ricerca Industriale Energia e Ambiente (CIRI EA) - Alma Mater Studiorum, University of Bologna, Bologna, Italy; Istituto Nazionale Biostrutture e Biosistemi (INBB), Roma, Italy. 8. Department of Medical and Surgical Sciences-DIMEC, Sant'Orsola Malpighi Hospital, Alma Mater Studiorum, University of Bologna, Bologna, Italy. 9. Department of Chemistry "Giacomo Ciamician" - Alma Mater Studiorum, University of Bologna, Bologna, Italy; Centro Interdipartimentale di Ricerca Industriale Energia e Ambiente (CIRI EA) - Alma Mater Studiorum, University of Bologna, Bologna, Italy; Istituto Nazionale Biostrutture e Biosistemi (INBB), Roma, Italy. 10. Department of Chemistry "Giacomo Ciamician" - Alma Mater Studiorum, University of Bologna, Bologna, Italy; Istituto Nazionale Biostrutture e Biosistemi (INBB), Roma, Italy.
Abstract
BACKGROUND AND AIMS: Oxidized LDL (oxLDL) or pro-inflammatory stimuli lead to increased oxidative stress linked to endothelial dysfunction and atherosclerosis. The oxLDL receptor-1 (LOX1) is elevated within atheromas and cholesterol-lowering statins inhibit LOX1 expression. Berberine (BBR), an alkaloid extracted from plants of gender Berberis, has lipid-lowering and anti-inflammatory activity. However, its role in regulating LOX1-mediated signaling is still unknown. The aim of this study was to investigate the effect of BBR on oxLDL- and TNFα-induced endothelial dysfunction in human umbilical vein endothelial cells (HUVECs) and to compare it with that of lovastatin (LOVA). METHODS AND RESULTS: Cytotoxicity was determined by lactate dehydrogenase assay. Antioxidant capacity was measured with chemiluminescent and fluorescent method and intracellular ROS levels through a fluorescent dye. Gene and protein expression levels were assayed by qRT-PCR and western blot, respectively. HUVECs exposure to oxLDL (30 μg/ml) or TNFα (10 ng/ml) for 24 h led to a significant increase in LOX1 expression, effect abrogated by BBR (5 μM) and LOVA (5 μM). BBR but not LOVA treatment abolished the TNFα-induced cytotoxicity and restored the activation of Akt signaling. In spite of a low direct antioxidant capacity, both compounds reduced intracellular ROS levels generated by treatment of TNFα but only BBR inhibited NOX2 expression, MAPK/Erk1/2 signaling and subsequent NF-κB target genes VCAM and ICAM expression, induced by TNFα. CONCLUSIONS: These findings demonstrated for the first time that BBR could prevent the oxLDL and TNFα - induced LOX1 expression and oxidative stress, key events that lead to NOX, MAPK/Erk1/2 and NF-κB activation linked to endothelial dysfunction. CHEMICAL COMPOUNDS STUDIED IN THIS ARTICLE: Berberine (PubChem CID: 2353); Lovastatin (PubChem CID: 53232).
BACKGROUND AND AIMS: Oxidized LDL (oxLDL) or pro-inflammatory stimuli lead to increased oxidative stress linked to endothelial dysfunction and atherosclerosis. The oxLDL receptor-1 (LOX1) is elevated within atheromas and cholesterol-lowering statins inhibit LOX1 expression. Berberine (BBR), an alkaloid extracted from plants of gender Berberis, has lipid-lowering and anti-inflammatory activity. However, its role in regulating LOX1-mediated signaling is still unknown. The aim of this study was to investigate the effect of BBR on oxLDL- and TNFα-induced endothelial dysfunction in human umbilical vein endothelial cells (HUVECs) and to compare it with that of lovastatin (LOVA). METHODS AND RESULTS:Cytotoxicity was determined by lactate dehydrogenase assay. Antioxidant capacity was measured with chemiluminescent and fluorescent method and intracellular ROS levels through a fluorescent dye. Gene and protein expression levels were assayed by qRT-PCR and western blot, respectively. HUVECs exposure to oxLDL (30 μg/ml) or TNFα (10 ng/ml) for 24 h led to a significant increase in LOX1 expression, effect abrogated by BBR (5 μM) and LOVA (5 μM). BBR but not LOVA treatment abolished the TNFα-induced cytotoxicity and restored the activation of Akt signaling. In spite of a low direct antioxidant capacity, both compounds reduced intracellular ROS levels generated by treatment of TNFα but only BBR inhibited NOX2 expression, MAPK/Erk1/2 signaling and subsequent NF-κB target genes VCAM and ICAM expression, induced by TNFα. CONCLUSIONS: These findings demonstrated for the first time that BBR could prevent the oxLDL and TNFα - induced LOX1 expression and oxidative stress, key events that lead to NOX, MAPK/Erk1/2 and NF-κB activation linked to endothelial dysfunction. CHEMICAL COMPOUNDS STUDIED IN THIS ARTICLE: Berberine (PubChem CID: 2353); Lovastatin (PubChem CID: 53232).
Authors: Nur Aqilah Kamaruddin; Muhammad Nazrul Hakim Abdullah; Jun Jie Tan; Vuanghao Lim; Lai Yen Fong; Siti Aisyah Abd Ghafar; Yoke Keong Yong Journal: Evid Based Complement Alternat Med Date: 2022-04-16 Impact factor: 2.650