BACKGROUND M1 polarization of macrophages is an important pathological process in myocardial ischemia reperfusion injury, which is the major obstacle for the treatment of acute myocardial infarction. Currently, the st...BACKGROUND M1 polarization of macrophages is an important pathological process in myocardial ischemia reperfusion injury, which is the major obstacle for the treatment of acute myocardial infarction. Currently, the strategies and mechanisms of inhibiting M1 polarization are poorly explored. This study aims to investigate the role of soluble death receptor 5-Fc(s DR5-Fc) in regulating M1 polarization of macrophages under extreme conditions and explore the mechanisms from the aspect of glycolysis.METHODS Extreme conditions were induced in RAW264.7 cells. Real-time quantitative polymerase chain reaction and western blot were used to detect the expression of m RNA and proteins, respectively. Cell counting kit-8 was used to investigate the proliferation activity of cells. Expression levels of inflammatory cytokines were determined by enzyme-linked immunosorbent assay.RESULTS We found that s DR5-Fc rescues the proliferation of macrophages under extreme conditions, including nutrition deficiency, excessive peroxide, and ultraviolet irradiation. In addition, administration of s DR5-Fc inhibits the M1 polarization of macrophages induced by lipopolysaccharide(LPS) and interferon-gamma(IFN-γ), as the expression of M1 polarization markers CD86, CXC motif chemokine ligand 10, matrix metalloproteinase 9, and tumor necrosis factor-α, as well as the secretion of inflammatory factors interleukin(IL)-1β and IL-6, were significantly decreased. By further investigation of the mechanisms, the results showed that s DR5-Fc can recover the LPS and IFN-γ induced p H reduction, lactic acid elevation, and increased expression of hexokinase 2 and glucose transporter 1, which were markers of glycolysis in macrophages.CONCLUSIONS s DR5-Fc inhibits the M1 polarization of macrophages by blocking the glycolysis, which provides a new direction for the development of strategies in the treatment of myocardial ischemia reperfusion injury.展开更多
Objective: To investigate the therapeutic effect of a novel suicide gene system, yeast cytosine deaminase (YCD)/5-Fluorocytosine(5-FC), on P388/DBA murine leukemia model. Methods: P388-YCD-eGFP clone was selected afte...Objective: To investigate the therapeutic effect of a novel suicide gene system, yeast cytosine deaminase (YCD)/5-Fluorocytosine(5-FC), on P388/DBA murine leukemia model. Methods: P388-YCD-eGFP clone was selected after retrovirus transduction by limited dilution. P388-eGFP and wild type (wt) P388 cells were used as controls. (1) Tumorigenesis study: DBA mice were inoculated intravenously with P388-YCD-eGFP, P388-eGFP or wt P388 cells at a dosage of 5×10~6 per mouse (n=5). (2) Therapeutic effect study of 5-FC: After inoculation with P388-YCD-eGFP, P388-eGFP or wt P388 cells (n=5), 5-FC was administered at a dosage of 5 μmol/d in each mouse for 2 weeks. 20 μmol/d 5-FC were administered on the 8~ th day in case 5 μmol/d did not work. The percentage of eGFP^+ cells was detected by flow cytometry, frozen section or HE section. Results: Mice in YCD, eGFP and wt P388 group developed leukemia and survived (8.0±1.0) d, (7.6±0.89) d and (7.8±1.64) d (P>0.05), respectively. After being treated with 5-FC for 2 weeks, mice in YCD and eGFP^+ groups survived (13.2±1.79) d and (8.4±0.58) d (P<0.05). When 20 μmol/d 5-FC was administered, the percentage of eGFP^+ cells dropped from 21.9%±16.09% to 5.4%±5.6%, as well detected by FACS within 24h after treatment. Conclusion: 5-FC can eliminate YCD gene modified cells in vivo when cells are inoculated intravenously. YCD is an efficient suicide gene system in vivo.展开更多
基金supported by the National Natural Science Foundation of Beijing, China (No.7212027 & No.7214223)National Key Research and Development Program of China (2017YFC0908800)the Beijing Municipal Health Commission (PXM2020_026272_000002 & PXM2020_026272_000014)。
文摘BACKGROUND M1 polarization of macrophages is an important pathological process in myocardial ischemia reperfusion injury, which is the major obstacle for the treatment of acute myocardial infarction. Currently, the strategies and mechanisms of inhibiting M1 polarization are poorly explored. This study aims to investigate the role of soluble death receptor 5-Fc(s DR5-Fc) in regulating M1 polarization of macrophages under extreme conditions and explore the mechanisms from the aspect of glycolysis.METHODS Extreme conditions were induced in RAW264.7 cells. Real-time quantitative polymerase chain reaction and western blot were used to detect the expression of m RNA and proteins, respectively. Cell counting kit-8 was used to investigate the proliferation activity of cells. Expression levels of inflammatory cytokines were determined by enzyme-linked immunosorbent assay.RESULTS We found that s DR5-Fc rescues the proliferation of macrophages under extreme conditions, including nutrition deficiency, excessive peroxide, and ultraviolet irradiation. In addition, administration of s DR5-Fc inhibits the M1 polarization of macrophages induced by lipopolysaccharide(LPS) and interferon-gamma(IFN-γ), as the expression of M1 polarization markers CD86, CXC motif chemokine ligand 10, matrix metalloproteinase 9, and tumor necrosis factor-α, as well as the secretion of inflammatory factors interleukin(IL)-1β and IL-6, were significantly decreased. By further investigation of the mechanisms, the results showed that s DR5-Fc can recover the LPS and IFN-γ induced p H reduction, lactic acid elevation, and increased expression of hexokinase 2 and glucose transporter 1, which were markers of glycolysis in macrophages.CONCLUSIONS s DR5-Fc inhibits the M1 polarization of macrophages by blocking the glycolysis, which provides a new direction for the development of strategies in the treatment of myocardial ischemia reperfusion injury.
基金Supported in part by grants from"Outstanding physicianProgram" of public Health Department of ShanghaiMunicipal Government,China(98BR0 2 9),National NaturalScience Foundation of China(30 170 379
文摘Objective: To investigate the therapeutic effect of a novel suicide gene system, yeast cytosine deaminase (YCD)/5-Fluorocytosine(5-FC), on P388/DBA murine leukemia model. Methods: P388-YCD-eGFP clone was selected after retrovirus transduction by limited dilution. P388-eGFP and wild type (wt) P388 cells were used as controls. (1) Tumorigenesis study: DBA mice were inoculated intravenously with P388-YCD-eGFP, P388-eGFP or wt P388 cells at a dosage of 5×10~6 per mouse (n=5). (2) Therapeutic effect study of 5-FC: After inoculation with P388-YCD-eGFP, P388-eGFP or wt P388 cells (n=5), 5-FC was administered at a dosage of 5 μmol/d in each mouse for 2 weeks. 20 μmol/d 5-FC were administered on the 8~ th day in case 5 μmol/d did not work. The percentage of eGFP^+ cells was detected by flow cytometry, frozen section or HE section. Results: Mice in YCD, eGFP and wt P388 group developed leukemia and survived (8.0±1.0) d, (7.6±0.89) d and (7.8±1.64) d (P>0.05), respectively. After being treated with 5-FC for 2 weeks, mice in YCD and eGFP^+ groups survived (13.2±1.79) d and (8.4±0.58) d (P<0.05). When 20 μmol/d 5-FC was administered, the percentage of eGFP^+ cells dropped from 21.9%±16.09% to 5.4%±5.6%, as well detected by FACS within 24h after treatment. Conclusion: 5-FC can eliminate YCD gene modified cells in vivo when cells are inoculated intravenously. YCD is an efficient suicide gene system in vivo.
