BACKGROUND： Modern pharmacological studies have shown that Ginsenoside Rgl is one of the active components of ginseng that promote intelligence in the nervous system. Ginsenoside Rgl can improve memory and learning i...BACKGROUND： Modern pharmacological studies have shown that Ginsenoside Rgl is one of the active components of ginseng that promote intelligence in the nervous system. Ginsenoside Rgl can improve memory and learning in mouse models of β-amyloid protein （Aβ）-induced dementia. OBJECTIVE： To investigate whether effects of Ginsenoside Rgl against Aβ are associated with activity of nuclear factor-kappa B （NF-κB）. DESIGN, TIME AND SETTING： The randomized performed at the DME Center, Institute of Clinica controlled, cell biological experiment was Pharmacology, Guangzhou University of Chinese Medicine, China from July 2005 to May 2006. MATERIALS： Beta-amyloid fragment 25-35 （Aβ25-35） was supplied by the Neural Biochemical Laboratory, Xuanwu Hospital, Capital Medical University, China. Ginsenoside Rgl was obtained from National Institute for the Control of Pharmaceutical and Biological Products, China. Rabbit anti-rat NF-κB p65 antibody was purchased from Santa Cruz Biotechnology, USA. METHODS： Hippocampal neurons and cortical astrocytes of neonatal Sprague Dawley rats were harvested and treated with various concentrations （0, 5, 10, 20, and 40 μmol/L） of Aβ for 6, 12, and 24 hours to establish cellular models of Alzheimer＇s disease. Cellular models were pretreated with various concentrations of Ginsenoside Rgl （1,2, 4, 8, and 16 μmol/L）. According to cell morphology and activity, the following conditions were selected： 40 μmol/L Aβ for 24 hours, as well as 2, 4, and 8 μmol/L Ginsenoside Rg1. NF-κB activity was observed using immunofluorescence and cytochemical staining. MAIN OUTCOME MEASURES： Morphology and viability of hippocampal neurons and cortical astrocytes, and activities of NF-κB were measured. RESULTS： Hippocampal neuron activity was significantly greater in the normal and 2 and 4 μmol/L Ginsenoside Rgl groups compared with the model group （P 〈 0.05）. Astrocyte activity was significantly greater in the normal, 1,2, 4, 8, and 16 μmol/L Ginsenoside Rgl groups compared with the model group （P 〈 0.05）. NF-κB activity of hippocampal neurons was significantly greater in the normal, 2, 4, and 8 μmol/L Ginsenoside Rgl groups compared with the model group （P 〈 0.01）. NF-κB activity of astrocytes was significantly less in the normal, 2, 4, and 8 μmol/L Ginsenoside Rgl groups compared with the model group （P 〈 0.01 or P 〈 0.05）. No significant difference in NF-κB activity was determined between the 2 μmol/L Ginsenoside Rgl and normal groups （P 〉 0.05）. CONCLUSION： Ginsenoside Rgl protected neural cells by upregulating NF-κB activity in neurons and downregulating NF-κB activity in astrocytes. Ginsenoside Rgl （2 μmol/L） maintained cell activity and NF-κB activity at normal levels.展开更多
OBJECTIVE To investigate the CKLF1 mediated expression of microglia/macrophage phenotypes in vitro and in vivo,discussing the involved pathway.METHODS In vitro,primary microglia isolated from mice cortex were used to ...OBJECTIVE To investigate the CKLF1 mediated expression of microglia/macrophage phenotypes in vitro and in vivo,discussing the involved pathway.METHODS In vitro,primary microglia isolated from mice cortex were used to study the effects of CKLF1 by qPCR analysis and immunofluorescence staining.In vivo,WT C57 and CKLF1 deficient mice were used to explore the effects of CKLF1.TTC staining,MRI and Nissl staining were applied to examine the infarction or neuron loss.Zea longa test was used to evaluate the neurological deficit of mice.Western blotting was used to investigate the changes of specific protein and discuss the involved pathway.We also used qPCR analysis and immunofluorescence staining for polarization markers to determine the effects of CKLF1.RESULTS CKLF1 could drive primary microglia to M1 phenotype for 24 h stimulation in primary microglia.In mice transient ischemic stroke model,CKLF1 attenuated ischemic injury,and accompanied by promoting microglia/macrophage toward M1 polarization.Increased expression of pro-inflammatory cytokines and decreased expression of neurotropic factors and anti-inflammatory cytokines were observed in mice subjected to cerebral ischemia with C27.Moreover,NF-κB activation enhancement was detected in C27 modulated M1 polarization effects.CONCLUSION CKLF1 is an important mediator of driving M1 phenotype of microglia/macrophage at early stage of cerebral ischemic injury,contributing to aggravation of cerebral ischemia injury,which closely related to microglia/macrophage M1 polarization guided inflammatory response.Targeting CKLF1 has the potential to treat ischemic stroke.展开更多
基金the Natural Science Foundation of Guangdong Province,No. 031479
文摘BACKGROUND： Modern pharmacological studies have shown that Ginsenoside Rgl is one of the active components of ginseng that promote intelligence in the nervous system. Ginsenoside Rgl can improve memory and learning in mouse models of β-amyloid protein （Aβ）-induced dementia. OBJECTIVE： To investigate whether effects of Ginsenoside Rgl against Aβ are associated with activity of nuclear factor-kappa B （NF-κB）. DESIGN, TIME AND SETTING： The randomized performed at the DME Center, Institute of Clinica controlled, cell biological experiment was Pharmacology, Guangzhou University of Chinese Medicine, China from July 2005 to May 2006. MATERIALS： Beta-amyloid fragment 25-35 （Aβ25-35） was supplied by the Neural Biochemical Laboratory, Xuanwu Hospital, Capital Medical University, China. Ginsenoside Rgl was obtained from National Institute for the Control of Pharmaceutical and Biological Products, China. Rabbit anti-rat NF-κB p65 antibody was purchased from Santa Cruz Biotechnology, USA. METHODS： Hippocampal neurons and cortical astrocytes of neonatal Sprague Dawley rats were harvested and treated with various concentrations （0, 5, 10, 20, and 40 μmol/L） of Aβ for 6, 12, and 24 hours to establish cellular models of Alzheimer＇s disease. Cellular models were pretreated with various concentrations of Ginsenoside Rgl （1,2, 4, 8, and 16 μmol/L）. According to cell morphology and activity, the following conditions were selected： 40 μmol/L Aβ for 24 hours, as well as 2, 4, and 8 μmol/L Ginsenoside Rg1. NF-κB activity was observed using immunofluorescence and cytochemical staining. MAIN OUTCOME MEASURES： Morphology and viability of hippocampal neurons and cortical astrocytes, and activities of NF-κB were measured. RESULTS： Hippocampal neuron activity was significantly greater in the normal and 2 and 4 μmol/L Ginsenoside Rgl groups compared with the model group （P 〈 0.05）. Astrocyte activity was significantly greater in the normal, 1,2, 4, 8, and 16 μmol/L Ginsenoside Rgl groups compared with the model group （P 〈 0.05）. NF-κB activity of hippocampal neurons was significantly greater in the normal, 2, 4, and 8 μmol/L Ginsenoside Rgl groups compared with the model group （P 〈 0.01）. NF-κB activity of astrocytes was significantly less in the normal, 2, 4, and 8 μmol/L Ginsenoside Rgl groups compared with the model group （P 〈 0.01 or P 〈 0.05）. No significant difference in NF-κB activity was determined between the 2 μmol/L Ginsenoside Rgl and normal groups （P 〉 0.05）. CONCLUSION： Ginsenoside Rgl protected neural cells by upregulating NF-κB activity in neurons and downregulating NF-κB activity in astrocytes. Ginsenoside Rgl （2 μmol/L） maintained cell activity and NF-κB activity at normal levels.
基金National Natural Science Foundation of China(81730096U1402221)+8 种基金National Mega-projectfor Innovative Drugs(2018ZX09711001-002-0072018ZX09711001-003-0052018ZX09711001-009-013)CAMS Innovation Fund for MedicalSciences(CIFMS)(2016-I2M-1-004)Beijing Key Laboratory of New Drug Mechanisms and Pharmacological Evaluation Study(BZ0150)the State Key Laboratory Fund Open Project(GTZK201610)China Postdoctoral Science Foundation(2013M540066)Project of NDRC and State Administration of Traditional Chinese Medicine(60011000)and Hunan Provincial Key Laboratory for Standardization of Important Chinese Herbal Pieces(BG201701,4981-0901020)
文摘OBJECTIVE To investigate the CKLF1 mediated expression of microglia/macrophage phenotypes in vitro and in vivo,discussing the involved pathway.METHODS In vitro,primary microglia isolated from mice cortex were used to study the effects of CKLF1 by qPCR analysis and immunofluorescence staining.In vivo,WT C57 and CKLF1 deficient mice were used to explore the effects of CKLF1.TTC staining,MRI and Nissl staining were applied to examine the infarction or neuron loss.Zea longa test was used to evaluate the neurological deficit of mice.Western blotting was used to investigate the changes of specific protein and discuss the involved pathway.We also used qPCR analysis and immunofluorescence staining for polarization markers to determine the effects of CKLF1.RESULTS CKLF1 could drive primary microglia to M1 phenotype for 24 h stimulation in primary microglia.In mice transient ischemic stroke model,CKLF1 attenuated ischemic injury,and accompanied by promoting microglia/macrophage toward M1 polarization.Increased expression of pro-inflammatory cytokines and decreased expression of neurotropic factors and anti-inflammatory cytokines were observed in mice subjected to cerebral ischemia with C27.Moreover,NF-κB activation enhancement was detected in C27 modulated M1 polarization effects.CONCLUSION CKLF1 is an important mediator of driving M1 phenotype of microglia/macrophage at early stage of cerebral ischemic injury,contributing to aggravation of cerebral ischemia injury,which closely related to microglia/macrophage M1 polarization guided inflammatory response.Targeting CKLF1 has the potential to treat ischemic stroke.
