In the developing and adult brain, neurotrophic growth factors support the growth and protec tion of dopaminergic neuronal systems. Recently, links between impaired neurotrophin support of dopamine (DA) neurons has be...In the developing and adult brain, neurotrophic growth factors support the growth and protec tion of dopaminergic neuronal systems. Recently, links between impaired neurotrophin support of dopamine (DA) neurons has been described in Parkinson’s Disease (PD). Fibro- blast growth factor (FGF) has a unique association with DA neurons in that FGF signaling is vitally important for the development and protection of adult DA neurons. We assessed the role of substantia nigra (SN)-expressed FGFs in the nigrostriatal dopaminergic system using a transgenic mouse, th-fgfr1(tk-). In these mice, generated by expression of dominant negative FGFR1(TK-) from the tyrosine hydroxylase (TH) gene promoter, reduced FGF signaling results in smaller and less dense adult nigrostriatal DA neurons, similar to what is observed in PD. With unilateral 6-hydroxydopamine (6-OHDA) lesions, th-fgfr1(tk-) mice exhibited extensive unilateral nigrostriatal damage with robust spontaneous (non-drug induced) asymmetrical turning and a decreased latency to remain on the accelerating rotarod. L-DOPA remains the gold standard for PD therapy despite debilitating hyperkinetic and dyskinetic side effects. The nicotinic acetylcholine system has recently been targeted as an alternative system to combat PD motor symptoms. Nicotine effectively stimulates dopaminergic transmission in the nigrostriatal pathway and mediates movement. Using unilaterally lesioned th-fgfr1(tk-) mice, long term (11 day) oral administration of nicotine increased spontaneous bidirectional turning and increased the latency before falling from the accelerating rotarod. In a separate analysis, L-DOPA treatment reversed directionality of rotation and further deepened motor discoordination, suggesting activation of hypersensitive postsynaptic DA receptors in the denervated striata. These results in a transgenic model of PD provide insights into the cellular mechanisms underlying L-DOPA and nicotinic therapies and offer further evidence of nicotine’s capacity to facilitate movement and enhance motor coordination in PD.展开更多
目的研究成纤维细胞生长因子18(FGF18)是否能诱导体外分离培养的人牙龈成纤维细胞(HGFs)向成骨样细胞分化,并探究其成骨机制。方法组织块法分离培养HGFs并鉴定。取第3代HGFs,分为实验组和对照组。实验组加入FGF18和L-DMEM、对照组加入L-...目的研究成纤维细胞生长因子18(FGF18)是否能诱导体外分离培养的人牙龈成纤维细胞(HGFs)向成骨样细胞分化,并探究其成骨机制。方法组织块法分离培养HGFs并鉴定。取第3代HGFs,分为实验组和对照组。实验组加入FGF18和L-DMEM、对照组加入L-DMEM。噻唑蓝(MTT)法检测不同浓度FGF18(0、0.01、0.02、0.04、0.06 mg/L)对HGFs增殖影响;碱性磷酸酶(ALP)和茜素红染色检测成骨能力和矿化能力;RT-PCR、免疫细胞化学染色及Western blot检测成骨相关基因、蛋白和BMP信号通路中BMP2基因和蛋白表达情况。结果与对照组比较,实验组培养3、5、7、9、11 d均可促进HGFs增殖(P<0.05);培养14、21 d ALP活性、矿物盐沉积均增高(P<0.05),ALP、OPN、OCN及BMP信号通路中BMP2 mRNA表达均明显增高(P<0.01)。培养21 d OPN、OCN及BMP2蛋白表达较培养14 d明显增高(P<0.01)。结论FGF18能促进HGFs增殖,诱导HGFs向功能性成骨样细胞分化,其成骨机制与上调BMP2有关。展开更多
焦虑症是高发的心理疾病,研究表明肥胖是导致焦虑症的重要危险因素。该研究探讨竹节参总皂苷(saponins from Panax japonicus,SPJ)对高脂饮食小鼠焦虑症的改善作用及机制。将50只C57BL/6J小鼠随机分为正常组、高脂饮食组、SPJ低剂量组、...焦虑症是高发的心理疾病,研究表明肥胖是导致焦虑症的重要危险因素。该研究探讨竹节参总皂苷(saponins from Panax japonicus,SPJ)对高脂饮食小鼠焦虑症的改善作用及机制。将50只C57BL/6J小鼠随机分为正常组、高脂饮食组、SPJ低剂量组、SPJ高剂量组。第12周从高脂饮食组取6只小鼠,分为对照组、外源性给予成纤维细胞生长因子21(FGF21)组。旷场和高架十字迷宫实验评价小鼠焦虑样行为;苏木素-伊红(HE)染色和油红O染色观察肝脏、脂肪的病理变化;葡萄糖耐量实验和胰岛素耐量实验评估小鼠糖代谢水平;蛋白免疫印迹法检测肝脏和皮层中FGF21及下游相关蛋白和大脑皮层中脑源性神经营养因子(BDNF)、突触后膜蛋白4(DLG4)和突触素(SYP)表达;实时荧光定量PCR(qPCR)检测肝脏和皮层中FGF21及相关受体基因表达;免疫荧光检测大脑皮层神经元激活物(c-Fos)、FGF21、FGF21辅助受体β-klotho的表达。结果显示,SPJ可显著改善高脂饮食小鼠在高架十字迷宫的开臂区及旷场中心区的活动频率,并上调BDNF、DLG4和SYP表达,有效改善高脂饮食小鼠的焦虑样行为。与正常组相比,高脂饮食组小鼠的肝脏及大脑皮层中FGF21的表达上调,成纤维细胞生长因子受体1(FGFR1)与β-klotho的表达显著下降,提示高脂饮食小鼠呈FGF21抵抗状态;SPJ可显著上调高脂饮食小鼠β-klotho水平,逆转FGF21抵抗。进一步与外源性给予FGF21进行对比,显示SPJ激活大脑皮层区域与其一致,同时SPJ能促进大脑皮层中c-Fos与β-klotho阳性细胞数量及共定位。综上所述,SPJ可有效改善高脂饮食小鼠的焦虑样行为,其机制与上调大脑β-klotho表达,逆转FGF21抵抗,进而激活大脑皮层、杏仁核神经元有关。展开更多
文摘In the developing and adult brain, neurotrophic growth factors support the growth and protec tion of dopaminergic neuronal systems. Recently, links between impaired neurotrophin support of dopamine (DA) neurons has been described in Parkinson’s Disease (PD). Fibro- blast growth factor (FGF) has a unique association with DA neurons in that FGF signaling is vitally important for the development and protection of adult DA neurons. We assessed the role of substantia nigra (SN)-expressed FGFs in the nigrostriatal dopaminergic system using a transgenic mouse, th-fgfr1(tk-). In these mice, generated by expression of dominant negative FGFR1(TK-) from the tyrosine hydroxylase (TH) gene promoter, reduced FGF signaling results in smaller and less dense adult nigrostriatal DA neurons, similar to what is observed in PD. With unilateral 6-hydroxydopamine (6-OHDA) lesions, th-fgfr1(tk-) mice exhibited extensive unilateral nigrostriatal damage with robust spontaneous (non-drug induced) asymmetrical turning and a decreased latency to remain on the accelerating rotarod. L-DOPA remains the gold standard for PD therapy despite debilitating hyperkinetic and dyskinetic side effects. The nicotinic acetylcholine system has recently been targeted as an alternative system to combat PD motor symptoms. Nicotine effectively stimulates dopaminergic transmission in the nigrostriatal pathway and mediates movement. Using unilaterally lesioned th-fgfr1(tk-) mice, long term (11 day) oral administration of nicotine increased spontaneous bidirectional turning and increased the latency before falling from the accelerating rotarod. In a separate analysis, L-DOPA treatment reversed directionality of rotation and further deepened motor discoordination, suggesting activation of hypersensitive postsynaptic DA receptors in the denervated striata. These results in a transgenic model of PD provide insights into the cellular mechanisms underlying L-DOPA and nicotinic therapies and offer further evidence of nicotine’s capacity to facilitate movement and enhance motor coordination in PD.
文摘目的研究成纤维细胞生长因子18(FGF18)是否能诱导体外分离培养的人牙龈成纤维细胞(HGFs)向成骨样细胞分化,并探究其成骨机制。方法组织块法分离培养HGFs并鉴定。取第3代HGFs,分为实验组和对照组。实验组加入FGF18和L-DMEM、对照组加入L-DMEM。噻唑蓝(MTT)法检测不同浓度FGF18(0、0.01、0.02、0.04、0.06 mg/L)对HGFs增殖影响;碱性磷酸酶(ALP)和茜素红染色检测成骨能力和矿化能力;RT-PCR、免疫细胞化学染色及Western blot检测成骨相关基因、蛋白和BMP信号通路中BMP2基因和蛋白表达情况。结果与对照组比较,实验组培养3、5、7、9、11 d均可促进HGFs增殖(P<0.05);培养14、21 d ALP活性、矿物盐沉积均增高(P<0.05),ALP、OPN、OCN及BMP信号通路中BMP2 mRNA表达均明显增高(P<0.01)。培养21 d OPN、OCN及BMP2蛋白表达较培养14 d明显增高(P<0.01)。结论FGF18能促进HGFs增殖,诱导HGFs向功能性成骨样细胞分化,其成骨机制与上调BMP2有关。
文摘焦虑症是高发的心理疾病,研究表明肥胖是导致焦虑症的重要危险因素。该研究探讨竹节参总皂苷(saponins from Panax japonicus,SPJ)对高脂饮食小鼠焦虑症的改善作用及机制。将50只C57BL/6J小鼠随机分为正常组、高脂饮食组、SPJ低剂量组、SPJ高剂量组。第12周从高脂饮食组取6只小鼠,分为对照组、外源性给予成纤维细胞生长因子21(FGF21)组。旷场和高架十字迷宫实验评价小鼠焦虑样行为;苏木素-伊红(HE)染色和油红O染色观察肝脏、脂肪的病理变化;葡萄糖耐量实验和胰岛素耐量实验评估小鼠糖代谢水平;蛋白免疫印迹法检测肝脏和皮层中FGF21及下游相关蛋白和大脑皮层中脑源性神经营养因子(BDNF)、突触后膜蛋白4(DLG4)和突触素(SYP)表达;实时荧光定量PCR(qPCR)检测肝脏和皮层中FGF21及相关受体基因表达;免疫荧光检测大脑皮层神经元激活物(c-Fos)、FGF21、FGF21辅助受体β-klotho的表达。结果显示,SPJ可显著改善高脂饮食小鼠在高架十字迷宫的开臂区及旷场中心区的活动频率,并上调BDNF、DLG4和SYP表达,有效改善高脂饮食小鼠的焦虑样行为。与正常组相比,高脂饮食组小鼠的肝脏及大脑皮层中FGF21的表达上调,成纤维细胞生长因子受体1(FGFR1)与β-klotho的表达显著下降,提示高脂饮食小鼠呈FGF21抵抗状态;SPJ可显著上调高脂饮食小鼠β-klotho水平,逆转FGF21抵抗。进一步与外源性给予FGF21进行对比,显示SPJ激活大脑皮层区域与其一致,同时SPJ能促进大脑皮层中c-Fos与β-klotho阳性细胞数量及共定位。综上所述,SPJ可有效改善高脂饮食小鼠的焦虑样行为,其机制与上调大脑β-klotho表达,逆转FGF21抵抗,进而激活大脑皮层、杏仁核神经元有关。
