Hamid A Bakshi1, Faruck Lukmanul Hakkim, Smitha Sam. 1. Department of Food Science and Human Nutrition, College of Applied science, A'Sharqiyah University, Ibra, Oman E-mail : hamidbakshi@asu.edu.om, hamid.bakshi@gmail.com.
Abstract
BACKGROUND: Crocus sativus and its major constituent crocin are well established to have anti-cancer properties in breast cancer cells (MCF-7). However the role of C. sativus extract (CSE) and crocin on caspase signaling mediated MCF-7 cell death at molecular level is remains unclear. In this study, we tried to unravel role of CSE and crocin on caspase mediated MCF-7 cells death and their in vivo preclinical toxicity profiling and immune stimulatory effect. MATERIALS AND METHODS: CSE extract was fractionated by HPLC and crocin was isolated and characterized by NMR, IR, and MS. MCF-7 cells were treated with both CSE and crocin and expression of Bcl-2 and Bax was assessed after 24 and 36 hours. Furthermore, caspase 3, caspase 8 and caspase 9 expression was determined by Western blotting after 24 hours of treatment. DNA fragmentation analysis was performed for genotoxicity of CSE and crocin in MCF-7 cells. The in vivo toxicity profile of CSE (300 mg/kg of b.wt) was investigated in normal Swiss albino mice. In addition, peritoneal macrophages were collected from crocin (1, 1.5 and 2 mg/kg body weight) treated mice and analyzed for ex vivo yeast phagocytosis. RESULTS: Immunoblot analysis revealed that there was time dependent decline in anti-apoptotic Bcl-2 with simultaneous upregulation of Bax in CSE and crocin treated MCF-7 cells. Further CSE and crocin treatment downregulated caspase 8 and 9 and cleaved the caspase 3 after 24 hours. Both CSE and crocin elicited considerable DNA damage in MCF-7 cells at each concentration tested. In vivo toxicity profile by histological studies revealed no observable histopathologic differences in the liver, kidney, spleen, lungs and heart in CSE treated and untreated groups. Crocin treatment elicited significant dose and time dependent ex vivo yeast phagocytosis by peritoneal macrophages. CONCLUSIONS: Our study delineated involvement of pro-apoptotic and caspase mediated MCF-7 cell death by CSE and crocin at the molecular level accompanied with extensive DNA damage. Further we found that normal swiss albino mice can tolerate the maximum dose of CSE. Crocin enhanced ex vivo macrophage yeast phagocytic ability.
BACKGROUND:Crocus sativus and its major constituent crocin are well established to have anti-cancer properties in breast cancer cells (MCF-7). However the role of C. sativus extract (CSE) and crocin on caspase signaling mediated MCF-7 cell death at molecular level is remains unclear. In this study, we tried to unravel role of CSE and crocin on caspase mediated MCF-7 cells death and their in vivo preclinical toxicity profiling and immune stimulatory effect. MATERIALS AND METHODS: CSE extract was fractionated by HPLC and crocin was isolated and characterized by NMR, IR, and MS. MCF-7 cells were treated with both CSE and crocin and expression of Bcl-2 and Bax was assessed after 24 and 36 hours. Furthermore, caspase 3, caspase 8 and caspase 9 expression was determined by Western blotting after 24 hours of treatment. DNA fragmentation analysis was performed for genotoxicity of CSE and crocin in MCF-7 cells. The in vivo toxicity profile of CSE (300 mg/kg of b.wt) was investigated in normal Swiss albino mice. In addition, peritoneal macrophages were collected from crocin (1, 1.5 and 2 mg/kg body weight) treated mice and analyzed for ex vivo yeast phagocytosis. RESULTS: Immunoblot analysis revealed that there was time dependent decline in anti-apoptotic Bcl-2 with simultaneous upregulation of Bax in CSE and crocin treated MCF-7 cells. Further CSE and crocin treatment downregulated caspase 8 and 9 and cleaved the caspase 3 after 24 hours. Both CSE and crocin elicited considerable DNA damage in MCF-7 cells at each concentration tested. In vivo toxicity profile by histological studies revealed no observable histopathologic differences in the liver, kidney, spleen, lungs and heart in CSE treated and untreated groups. Crocin treatment elicited significant dose and time dependent ex vivo yeast phagocytosis by peritoneal macrophages. CONCLUSIONS: Our study delineated involvement of pro-apoptotic and caspase mediated MCF-7 cell death by CSE and crocin at the molecular level accompanied with extensive DNA damage. Further we found that normal swiss albino mice can tolerate the maximum dose of CSE. Crocin enhanced ex vivo macrophage yeast phagocytic ability.
Authors: Kyriaki Hatziagapiou; Olti Nikola; Sofia Marka; Eleni Koniari; Eleni Kakouri; Maria-Eleftheria Zografaki; Sophie S Mavrikou; Charalabos Kanakis; Emmanouil Flemetakis; George P Chrousos; Spyridon Kintzios; George I Lambrou; Christina Kanaka-Gantenbein; Petros A Tarantilis Journal: Antioxidants (Basel) Date: 2022-05-28
Authors: Hamid A Bakshi; Mazhar S Al Zoubi; Faruck L Hakkim; Alaa A A Aljabali; Firas A Rabi; Amin A Hafiz; Khalid M Al-Batanyeh; Bahaa Al-Trad; Prawej Ansari; Mohamed M Nasef; Nitin B Charbe; Saurabh Satija; Meenu Mehta; Vijay Mishra; Gaurav Gupta; Salem Abobaker; Poonam Negi; Ibrahim M Azzouz; Ashref Ali K Dardouri; Harish Dureja; Parteek Prasher; Dinesh K Chellappan; Kamal Dua; Mateus Webba da Silva; Mohamed El Tanani; Paul A McCarron; Murtaza M Tambuwala Journal: Nutrients Date: 2020-06-26 Impact factor: 5.717
Authors: Ali Veisi; Ghaidafeh Akbari; Seyyed Ali Mard; Gholamreza Badfar; Vahid Zarezade; Mohammad Ali Mirshekar Journal: Iran J Basic Med Sci Date: 2020-01 Impact factor: 2.699