Wpływ feniksyny na białe i brunatne adipocyty oraz metabolizm i tkankę tłuszczową w otyłości
Type
Project
Date Issued
2023
Author
Discipline
biological sciences
Abstract (PL)
Research project objectives/Research hypothesis
Phoenixin is a peptide hormone which modulates reproduction, appetite and pain sensation. There is evidence indicating that biological effects of phoenixin are mediated trough GPR173 activation. Human data showed that circulating phoenixin level is positively associated with body mass index. In addition, recently we found that phoenixin modulates white adipogenesis. These results suggest that phoenixin may be involved in controlling body weight, as well as adipose tissue formations. While we showed that phoenixin modulates proliferation and differentiation of rodents’ white preadipocytes, the role of this peptide in controlling brown adipogenesis, functions of mature white fat cells and obesity is unknown. Based on the literature data and our preliminary studies, we hypothesize that phoenixin modulates brown adipogenesis and functions of mature white adipocytes. Therefore, the aim of this project is characterization of the effects of phoenixin on brown adipogenesis, as well as its effects on browning, glucose and lipid metabolism and endocrine functions of mature rodent and human white adipocytes. Moreover, we will study whether phoenixin modulates inflammatory response in white adipocytes. In addition the project aims to characterize the role of GPR173 and molecular mechanisms conferring biological effects of phoenixin in brown adipogenesis and white adipocytes functions. Finally, we will characterize the effects of phoenixin administration on metabolic status and adipose tissue in mice with high fat-induced obesity.
Research project methodology
To study the role of phoenixin in brown adipogenesis we will utilize rat brown primary preadipocytes. The effects of phoenixin on mature white adipocytes will be tested using fully differentiated mouse white 3T3-L1 adipocytes, as well as white rat and human primary adipocytes. Primary white and brown preadipocytes will be isolated from male Wistar rats (100 g, 5-6 weeks old). Preadipocytes will be differentiated using standard protocols. Tissue specimens of white human visceral adipose tissue will be obtained during routing surgery from male patients free of metabolic or endocrine diseases. The effects of phoenixin on cell proliferation, viability and apoptosis will be investigated using specific colorimetric tests. The role of phoenixin in controlling brown adipogenesis will be investigated by evaluating cell morphology, lipids accumulation, mitochondrial biogenesis, oxygen consumption and expression of adipogenic markers. Gene expression (mRNA) will be studied using real time PCR and protein production will be evaluated using Western blot. LPS and macrophages will be used to induce in vitro model of inflammation. The influence of phoenixin on glucose uptake and lipolysis will be assessed using specific tests and labeled glucose. ELISA tests, siRNA and specific pharmacological inhibitors will be used in order to study the role of GPR173 receptor and intracellular mechanisms conferring the effects of phoenixin on adipocytes. To study the effects of phoenixin on metabolic status and adipose tissue functions in obese mice will be fed high fat diet to induce obesity and treated with or without phoenixin for 4 weeks. Next Generation Sequencing (NGS) technique will be utilized to study a complex description of transcriptome differences in adipose tissue of obese and normal mice treated with or without phoenixin.
Expected impact of the research project on the development of science, civilization and society
The results of this project should answer the questions about the role of phoenixin in controlling brown adipogenesis, browning of white fat cells as well as its effects on endocrine activity and metabolism of mature rodent and human adipocytes. In addition, the project should elucidate the question whether phoenixin is able to modulate proinflammatory response in white adipocytes. The project should characterize molecular mechanism conferring the effects of phoenixin on brown adipogenesis and mature white adipocytes. The project should answer the question about the effects of phoenixin on metabolic status and adipose tissue in mice with experimentally induced obesity. We assume that the results of this project will improve our understanding of phoenixin functions in human and rodent fat cells and its potential role in therapy of obesity and obesity-related diseases.
Phoenixin is a peptide hormone which modulates reproduction, appetite and pain sensation. There is evidence indicating that biological effects of phoenixin are mediated trough GPR173 activation. Human data showed that circulating phoenixin level is positively associated with body mass index. In addition, recently we found that phoenixin modulates white adipogenesis. These results suggest that phoenixin may be involved in controlling body weight, as well as adipose tissue formations. While we showed that phoenixin modulates proliferation and differentiation of rodents’ white preadipocytes, the role of this peptide in controlling brown adipogenesis, functions of mature white fat cells and obesity is unknown. Based on the literature data and our preliminary studies, we hypothesize that phoenixin modulates brown adipogenesis and functions of mature white adipocytes. Therefore, the aim of this project is characterization of the effects of phoenixin on brown adipogenesis, as well as its effects on browning, glucose and lipid metabolism and endocrine functions of mature rodent and human white adipocytes. Moreover, we will study whether phoenixin modulates inflammatory response in white adipocytes. In addition the project aims to characterize the role of GPR173 and molecular mechanisms conferring biological effects of phoenixin in brown adipogenesis and white adipocytes functions. Finally, we will characterize the effects of phoenixin administration on metabolic status and adipose tissue in mice with high fat-induced obesity.
Research project methodology
To study the role of phoenixin in brown adipogenesis we will utilize rat brown primary preadipocytes. The effects of phoenixin on mature white adipocytes will be tested using fully differentiated mouse white 3T3-L1 adipocytes, as well as white rat and human primary adipocytes. Primary white and brown preadipocytes will be isolated from male Wistar rats (100 g, 5-6 weeks old). Preadipocytes will be differentiated using standard protocols. Tissue specimens of white human visceral adipose tissue will be obtained during routing surgery from male patients free of metabolic or endocrine diseases. The effects of phoenixin on cell proliferation, viability and apoptosis will be investigated using specific colorimetric tests. The role of phoenixin in controlling brown adipogenesis will be investigated by evaluating cell morphology, lipids accumulation, mitochondrial biogenesis, oxygen consumption and expression of adipogenic markers. Gene expression (mRNA) will be studied using real time PCR and protein production will be evaluated using Western blot. LPS and macrophages will be used to induce in vitro model of inflammation. The influence of phoenixin on glucose uptake and lipolysis will be assessed using specific tests and labeled glucose. ELISA tests, siRNA and specific pharmacological inhibitors will be used in order to study the role of GPR173 receptor and intracellular mechanisms conferring the effects of phoenixin on adipocytes. To study the effects of phoenixin on metabolic status and adipose tissue functions in obese mice will be fed high fat diet to induce obesity and treated with or without phoenixin for 4 weeks. Next Generation Sequencing (NGS) technique will be utilized to study a complex description of transcriptome differences in adipose tissue of obese and normal mice treated with or without phoenixin.
Expected impact of the research project on the development of science, civilization and society
The results of this project should answer the questions about the role of phoenixin in controlling brown adipogenesis, browning of white fat cells as well as its effects on endocrine activity and metabolism of mature rodent and human adipocytes. In addition, the project should elucidate the question whether phoenixin is able to modulate proinflammatory response in white adipocytes. The project should characterize molecular mechanism conferring the effects of phoenixin on brown adipogenesis and mature white adipocytes. The project should answer the question about the effects of phoenixin on metabolic status and adipose tissue in mice with experimentally induced obesity. We assume that the results of this project will improve our understanding of phoenixin functions in human and rodent fat cells and its potential role in therapy of obesity and obesity-related diseases.
Abstract (EN)
Research project objectives/Research hypothesis
Phoenixin is a peptide hormone which modulates reproduction, appetite and pain sensation. There is evidence indicating that biological effects of phoenixin are mediated trough GPR173 activation. Human data showed that circulating phoenixin level is positively associated with body mass index. In addition, recently we found that phoenixin modulates white adipogenesis. These results suggest that phoenixin may be involved in controlling body weight, as well as adipose tissue formations. While we showed that phoenixin modulates proliferation and differentiation of rodents’ white preadipocytes, the role of this peptide in controlling brown adipogenesis, functions of mature white fat cells and obesity is unknown. Based on the literature data and our preliminary studies, we hypothesize that phoenixin modulates brown adipogenesis and functions of mature white adipocytes. Therefore, the aim of this project is characterization of the effects of phoenixin on brown adipogenesis, as well as its effects on browning, glucose and lipid metabolism and endocrine functions of mature rodent and human white adipocytes. Moreover, we will study whether phoenixin modulates inflammatory response in white adipocytes. In addition the project aims to characterize the role of GPR173 and molecular mechanisms conferring biological effects of phoenixin in brown adipogenesis and white adipocytes functions. Finally, we will characterize the effects of phoenixin administration on metabolic status and adipose tissue in mice with high fat-induced obesity.
Research project methodology
To study the role of phoenixin in brown adipogenesis we will utilize rat brown primary preadipocytes. The effects of phoenixin on mature white adipocytes will be tested using fully differentiated mouse white 3T3-L1 adipocytes, as well as white rat and human primary adipocytes. Primary white and brown preadipocytes will be isolated from male Wistar rats (100 g, 5-6 weeks old). Preadipocytes will be differentiated using standard protocols. Tissue specimens of white human visceral adipose tissue will be obtained during routing surgery from male patients free of metabolic or endocrine diseases. The effects of phoenixin on cell proliferation, viability and apoptosis will be investigated using specific colorimetric tests. The role of phoenixin in controlling brown adipogenesis will be investigated by evaluating cell morphology, lipids accumulation, mitochondrial biogenesis, oxygen consumption and expression of adipogenic markers. Gene expression (mRNA) will be studied using real time PCR and protein production will be evaluated using Western blot. LPS and macrophages will be used to induce in vitro model of inflammation. The influence of phoenixin on glucose uptake and lipolysis will be assessed using specific tests and labeled glucose. ELISA tests, siRNA and specific pharmacological inhibitors will be used in order to study the role of GPR173 receptor and intracellular mechanisms conferring the effects of phoenixin on adipocytes. To study the effects of phoenixin on metabolic status and adipose tissue functions in obese mice will be fed high fat diet to induce obesity and treated with or without phoenixin for 4 weeks. Next Generation Sequencing (NGS) technique will be utilized to study a complex description of transcriptome differences in adipose tissue of obese and normal mice treated with or without phoenixin.
Expected impact of the research project on the development of science, civilization and society
The results of this project should answer the questions about the role of phoenixin in controlling brown adipogenesis, browning of white fat cells as well as its effects on endocrine activity and metabolism of mature rodent and human adipocytes. In addition, the project should elucidate the question whether phoenixin is able to modulate proinflammatory response in white adipocytes. The project should characterize molecular mechanism conferring the effects of phoenixin on brown adipogenesis and mature white adipocytes. The project should answer the question about the effects of phoenixin on metabolic status and adipose tissue in mice with experimentally induced obesity. We assume that the results of this project will improve our understanding of phoenixin functions in human and rodent fat cells and its potential role in therapy of obesity and obesity-related diseases.
Phoenixin is a peptide hormone which modulates reproduction, appetite and pain sensation. There is evidence indicating that biological effects of phoenixin are mediated trough GPR173 activation. Human data showed that circulating phoenixin level is positively associated with body mass index. In addition, recently we found that phoenixin modulates white adipogenesis. These results suggest that phoenixin may be involved in controlling body weight, as well as adipose tissue formations. While we showed that phoenixin modulates proliferation and differentiation of rodents’ white preadipocytes, the role of this peptide in controlling brown adipogenesis, functions of mature white fat cells and obesity is unknown. Based on the literature data and our preliminary studies, we hypothesize that phoenixin modulates brown adipogenesis and functions of mature white adipocytes. Therefore, the aim of this project is characterization of the effects of phoenixin on brown adipogenesis, as well as its effects on browning, glucose and lipid metabolism and endocrine functions of mature rodent and human white adipocytes. Moreover, we will study whether phoenixin modulates inflammatory response in white adipocytes. In addition the project aims to characterize the role of GPR173 and molecular mechanisms conferring biological effects of phoenixin in brown adipogenesis and white adipocytes functions. Finally, we will characterize the effects of phoenixin administration on metabolic status and adipose tissue in mice with high fat-induced obesity.
Research project methodology
To study the role of phoenixin in brown adipogenesis we will utilize rat brown primary preadipocytes. The effects of phoenixin on mature white adipocytes will be tested using fully differentiated mouse white 3T3-L1 adipocytes, as well as white rat and human primary adipocytes. Primary white and brown preadipocytes will be isolated from male Wistar rats (100 g, 5-6 weeks old). Preadipocytes will be differentiated using standard protocols. Tissue specimens of white human visceral adipose tissue will be obtained during routing surgery from male patients free of metabolic or endocrine diseases. The effects of phoenixin on cell proliferation, viability and apoptosis will be investigated using specific colorimetric tests. The role of phoenixin in controlling brown adipogenesis will be investigated by evaluating cell morphology, lipids accumulation, mitochondrial biogenesis, oxygen consumption and expression of adipogenic markers. Gene expression (mRNA) will be studied using real time PCR and protein production will be evaluated using Western blot. LPS and macrophages will be used to induce in vitro model of inflammation. The influence of phoenixin on glucose uptake and lipolysis will be assessed using specific tests and labeled glucose. ELISA tests, siRNA and specific pharmacological inhibitors will be used in order to study the role of GPR173 receptor and intracellular mechanisms conferring the effects of phoenixin on adipocytes. To study the effects of phoenixin on metabolic status and adipose tissue functions in obese mice will be fed high fat diet to induce obesity and treated with or without phoenixin for 4 weeks. Next Generation Sequencing (NGS) technique will be utilized to study a complex description of transcriptome differences in adipose tissue of obese and normal mice treated with or without phoenixin.
Expected impact of the research project on the development of science, civilization and society
The results of this project should answer the questions about the role of phoenixin in controlling brown adipogenesis, browning of white fat cells as well as its effects on endocrine activity and metabolism of mature rodent and human adipocytes. In addition, the project should elucidate the question whether phoenixin is able to modulate proinflammatory response in white adipocytes. The project should characterize molecular mechanism conferring the effects of phoenixin on brown adipogenesis and mature white adipocytes. The project should answer the question about the effects of phoenixin on metabolic status and adipose tissue in mice with experimentally induced obesity. We assume that the results of this project will improve our understanding of phoenixin functions in human and rodent fat cells and its potential role in therapy of obesity and obesity-related diseases.