The interaction between metabolic dysfunction and inflammation is central to the development of neurodegenerative diseases such as Alzheimer’s disease and Parkinson’s disease.Obesity-related conditions like type 2 d...The interaction between metabolic dysfunction and inflammation is central to the development of neurodegenerative diseases such as Alzheimer’s disease and Parkinson’s disease.Obesity-related conditions like type 2 diabetes and non-alcoholic fatty liver disease exacerbate this relationship.Peripheral lipid accumulation,particularly in the liver,initiates a cascade of inflammatory processes that extend to the brain,influencing critical metabolic regulatory regions.Ceramide and palmitate,key lipid components,along with lipid transporters lipocalin-2 and apolipoprotein E,contribute to neuroinflammation by disrupting blood–brain barrier integrity and promoting gliosis.Peripheral insulin resistance further exacerbates brain insulin resistance and neuroinflammation.Preclinical interventions targeting peripheral lipid metabolism and insulin signaling pathways have shown promise in reducing neuroinflammation in animal models.However,translating these findings to clinical practice requires further investigation into human subjects.In conclusion,metabolic dysfunction,peripheral inflammation,and insulin resistance are integral to neuroinflammation and neurodegeneration.Understanding these complex mechanisms holds potential for identifying novel therapeutic targets and improving outcomes for neurodegenerative diseases.展开更多
Background:Liver diseases are a major contributor to both morbidity and mortality.Conditional knockout animals are always produced through crossing floxed animals with a tissue-specific Cre animal.The use of floxed ra...Background:Liver diseases are a major contributor to both morbidity and mortality.Conditional knockout animals are always produced through crossing floxed animals with a tissue-specific Cre animal.The use of floxed rat resource has rapidly increased,but the liver-specific Cre rat lines for studying liver diseases and interested genes are limited,especially in a spatially and temporally restricted manner.Methods:RNA sequencing and real-time polymerase chain reaction(PCR)were used to screen and confirm the presence of liver-specific genes.Apoa4-Cre rats and Cyp2c11-Cre rats were produced by CRISPR/Cas9 knockin.Rosa26-imCherry rats were employed to hybridize with the Cre rats to obtain the Apoa4-Cre/Rosa26-imCherry and Cyp2c11-Cre/Rosa26-imCherry rats.The temporal and spatial patterns of Cre expression were determined by the observation of red fluorescence on tis-sue sections.Hematoxylin-eosin stain was used to evaluate the liver histopathologic changes.The blood biochemical analysis of several liver enzymes and liver lipid profile was performed to evaluate the liver function of Cre rats.Results:Apoa4 and Cyp2c11 were identified as two liver-specific genes.Apoa4-Cre and Cyp2c11-Cre rats were produced and hybridized with Rosa26-imCherry rats.The red fluorescence indicated that the Cre recombinases were specially expressed in the juvenile and adult liver and not in other organs of two hybridized rats.All the blood biochemical parameters except low-density lipoprotein(LDL)did not change signifi-cantly in the Cre rats.No histological alterations were detected in the livers of the Cre rats.Conclusions:Liver-specific Apoa4-Cre and Cyp2c11-Cre rats have been established successfully and could be used to study gene knockout,specifically in juvenile and adult liver.展开更多
目的探究强直性脊柱炎(ankylosing spondylitis,AS)模型小鼠和临床患者外周血单个核细胞(peripheralblood mononuclear cell,PBMC)中微小RNA-142-5p(miR-142-5p),细胞因子信号转导抑制因子1(suppressor ofcytokine signaling 1,SOCS1)m...目的探究强直性脊柱炎(ankylosing spondylitis,AS)模型小鼠和临床患者外周血单个核细胞(peripheralblood mononuclear cell,PBMC)中微小RNA-142-5p(miR-142-5p),细胞因子信号转导抑制因子1(suppressor ofcytokine signaling 1,SOCS1)mRNA表达及其对免疫功能的影响。方法通过实时荧光定量(quantitative real timePCR,qRT-PCR)检测2022年1月~2023年3月商丘市第一人民医院收治的30例确诊的AS患者(患者组)和30例健康体检者(健康组)的PBMC中miR-142-5p和SOCS1 mRNA水平。采用牛蛋白聚糖联合完全弗氏佐剂诱导AS小鼠模型,然后将小鼠分为对照组、模型组、阴性组和拮抗剂组。对照组和模型组小鼠尾静脉注射生理盐水,阴性组和拮抗剂组小鼠分别尾静脉注射NC-antagomir和miR-142-5p-antagomir。治疗2周后,分别评估各组小鼠的关节炎症状评分。通过苏木精伊红(hematoxylin eosin,HE)染色评价踝关节形态。采用ELISA法检测小鼠血清中Th1细胞因子干扰素γ(IFN-γ)、Th2细胞因子白细胞介素-4(IL-4)、Th17细胞因子白细胞介素-17(IL-17)和Treg细胞因子叉头盒蛋白P3(FOXP3)的表达水平。通过qRT-PCR和Western blot检测PBMC和踝关节组织中miR-142-5p,SOCS1,IFN-γ,IL-4,IL-17和FOXP3的mRNA和蛋白表达水平。结果与健康组比较,患者组PBMC中miR-142-5p水平升高(3.03±0.99 vs1.00±0.21),SOCS1 mRNA水平降低(0.41±0.09 vs 1.00±0.18),差异具有统计学意义(t=10.997,15.956,均P<0.001)。与对照组比较,模型组小鼠踝关节组织中miR-142-5p水平(4.00±0.52 vs 1.00±0.04)升高,IFN-γ和IL-17的mRNA和蛋白水平均升高,SOCS1,IL-4和FOXP3的mRNA和蛋白水平均降低,差异具有统计学意义(t=23.356,31.420,48.056,47.224,38.035,29.007,54.183,28.123,55.155,26.758,45.346,均P<0.05);关节炎症状评分升高(7.83±0.94 vs 0.00±0.00,t=22.212,P<0.05),踝关节结构破坏明显;血清中IFN-γ,IL-17水平以及IFN-γ/IL-4比值(0.81±0.08 vs 2.08±0.33)和IL-17/FOXP3比值(0.41±0.03 vs 1.27±0.10)均升高,差异具有统计学意义(t=15.382,35.779,15.412,35.130,均P<0.05)。与阴性组比较,拮抗剂组小鼠踝关节组织中miR-142-5p水平(1.47±0.10 vs 3.89±0.33)降低,IFN-γ和IL-17的mRNA和蛋白水平均降低,SOCS1,IL-4和FOXP3的mRNA和蛋白水平均升高,差异具有统计学意义(t=18.846,22.969,43.454,32.617,23.259,20.881,41.832,11.994,32.977,15.190,35.834,均P<0.05);关节炎症状评分降低(7.42±1.24 vs 2.75±0.75,t=13.233,P<0.05),踝关节形态明显改善;血清中IFN-γ,IL-17水平以及IFN-γ/IL-4比值(1.22±0.11 vs 1.91±0.19)和IL-17/FOXP3比值(0.69±0.05vs 1.23±0.12)均降低,差异具有统计学意义(t=8.688,22.972,8.377,22.007,均P<0.05)。结论miR-142-5p在AS中高表达,使用拮抗剂下调miR-142-5p可能通过上调SOCS1进而降低Th1/Th2和Th17/Treg比值,从而改善AS小鼠的免疫平衡并抑制AS的进展。展开更多
基金supported by a Presidential Postdoctoral Fellowship (021229-00001) from Nanyang Technological University,Singapore (to JZ)a Lee Kong Chian School of Medicine Dean’s Postdoctoral Fellowship (021207-00001) from NTU Singaporea Mistletoe Research Fellowship (022522-00001) from the Momental Foundaton,USA (to CHL)
文摘The interaction between metabolic dysfunction and inflammation is central to the development of neurodegenerative diseases such as Alzheimer’s disease and Parkinson’s disease.Obesity-related conditions like type 2 diabetes and non-alcoholic fatty liver disease exacerbate this relationship.Peripheral lipid accumulation,particularly in the liver,initiates a cascade of inflammatory processes that extend to the brain,influencing critical metabolic regulatory regions.Ceramide and palmitate,key lipid components,along with lipid transporters lipocalin-2 and apolipoprotein E,contribute to neuroinflammation by disrupting blood–brain barrier integrity and promoting gliosis.Peripheral insulin resistance further exacerbates brain insulin resistance and neuroinflammation.Preclinical interventions targeting peripheral lipid metabolism and insulin signaling pathways have shown promise in reducing neuroinflammation in animal models.However,translating these findings to clinical practice requires further investigation into human subjects.In conclusion,metabolic dysfunction,peripheral inflammation,and insulin resistance are integral to neuroinflammation and neurodegeneration.Understanding these complex mechanisms holds potential for identifying novel therapeutic targets and improving outcomes for neurodegenerative diseases.
基金CAMS Innovation Fund for Medical Sciences(CIFMS),Grant/Award Number:2021-I2M-1-035National Natural Science Foundation of China,Grant/Award Number:31970508。
文摘Background:Liver diseases are a major contributor to both morbidity and mortality.Conditional knockout animals are always produced through crossing floxed animals with a tissue-specific Cre animal.The use of floxed rat resource has rapidly increased,but the liver-specific Cre rat lines for studying liver diseases and interested genes are limited,especially in a spatially and temporally restricted manner.Methods:RNA sequencing and real-time polymerase chain reaction(PCR)were used to screen and confirm the presence of liver-specific genes.Apoa4-Cre rats and Cyp2c11-Cre rats were produced by CRISPR/Cas9 knockin.Rosa26-imCherry rats were employed to hybridize with the Cre rats to obtain the Apoa4-Cre/Rosa26-imCherry and Cyp2c11-Cre/Rosa26-imCherry rats.The temporal and spatial patterns of Cre expression were determined by the observation of red fluorescence on tis-sue sections.Hematoxylin-eosin stain was used to evaluate the liver histopathologic changes.The blood biochemical analysis of several liver enzymes and liver lipid profile was performed to evaluate the liver function of Cre rats.Results:Apoa4 and Cyp2c11 were identified as two liver-specific genes.Apoa4-Cre and Cyp2c11-Cre rats were produced and hybridized with Rosa26-imCherry rats.The red fluorescence indicated that the Cre recombinases were specially expressed in the juvenile and adult liver and not in other organs of two hybridized rats.All the blood biochemical parameters except low-density lipoprotein(LDL)did not change signifi-cantly in the Cre rats.No histological alterations were detected in the livers of the Cre rats.Conclusions:Liver-specific Apoa4-Cre and Cyp2c11-Cre rats have been established successfully and could be used to study gene knockout,specifically in juvenile and adult liver.
文摘目的探究强直性脊柱炎(ankylosing spondylitis,AS)模型小鼠和临床患者外周血单个核细胞(peripheralblood mononuclear cell,PBMC)中微小RNA-142-5p(miR-142-5p),细胞因子信号转导抑制因子1(suppressor ofcytokine signaling 1,SOCS1)mRNA表达及其对免疫功能的影响。方法通过实时荧光定量(quantitative real timePCR,qRT-PCR)检测2022年1月~2023年3月商丘市第一人民医院收治的30例确诊的AS患者(患者组)和30例健康体检者(健康组)的PBMC中miR-142-5p和SOCS1 mRNA水平。采用牛蛋白聚糖联合完全弗氏佐剂诱导AS小鼠模型,然后将小鼠分为对照组、模型组、阴性组和拮抗剂组。对照组和模型组小鼠尾静脉注射生理盐水,阴性组和拮抗剂组小鼠分别尾静脉注射NC-antagomir和miR-142-5p-antagomir。治疗2周后,分别评估各组小鼠的关节炎症状评分。通过苏木精伊红(hematoxylin eosin,HE)染色评价踝关节形态。采用ELISA法检测小鼠血清中Th1细胞因子干扰素γ(IFN-γ)、Th2细胞因子白细胞介素-4(IL-4)、Th17细胞因子白细胞介素-17(IL-17)和Treg细胞因子叉头盒蛋白P3(FOXP3)的表达水平。通过qRT-PCR和Western blot检测PBMC和踝关节组织中miR-142-5p,SOCS1,IFN-γ,IL-4,IL-17和FOXP3的mRNA和蛋白表达水平。结果与健康组比较,患者组PBMC中miR-142-5p水平升高(3.03±0.99 vs1.00±0.21),SOCS1 mRNA水平降低(0.41±0.09 vs 1.00±0.18),差异具有统计学意义(t=10.997,15.956,均P<0.001)。与对照组比较,模型组小鼠踝关节组织中miR-142-5p水平(4.00±0.52 vs 1.00±0.04)升高,IFN-γ和IL-17的mRNA和蛋白水平均升高,SOCS1,IL-4和FOXP3的mRNA和蛋白水平均降低,差异具有统计学意义(t=23.356,31.420,48.056,47.224,38.035,29.007,54.183,28.123,55.155,26.758,45.346,均P<0.05);关节炎症状评分升高(7.83±0.94 vs 0.00±0.00,t=22.212,P<0.05),踝关节结构破坏明显;血清中IFN-γ,IL-17水平以及IFN-γ/IL-4比值(0.81±0.08 vs 2.08±0.33)和IL-17/FOXP3比值(0.41±0.03 vs 1.27±0.10)均升高,差异具有统计学意义(t=15.382,35.779,15.412,35.130,均P<0.05)。与阴性组比较,拮抗剂组小鼠踝关节组织中miR-142-5p水平(1.47±0.10 vs 3.89±0.33)降低,IFN-γ和IL-17的mRNA和蛋白水平均降低,SOCS1,IL-4和FOXP3的mRNA和蛋白水平均升高,差异具有统计学意义(t=18.846,22.969,43.454,32.617,23.259,20.881,41.832,11.994,32.977,15.190,35.834,均P<0.05);关节炎症状评分降低(7.42±1.24 vs 2.75±0.75,t=13.233,P<0.05),踝关节形态明显改善;血清中IFN-γ,IL-17水平以及IFN-γ/IL-4比值(1.22±0.11 vs 1.91±0.19)和IL-17/FOXP3比值(0.69±0.05vs 1.23±0.12)均降低,差异具有统计学意义(t=8.688,22.972,8.377,22.007,均P<0.05)。结论miR-142-5p在AS中高表达,使用拮抗剂下调miR-142-5p可能通过上调SOCS1进而降低Th1/Th2和Th17/Treg比值,从而改善AS小鼠的免疫平衡并抑制AS的进展。
