目的研究葛根素对氧糖剥夺(oxygen and glucose deprivation,OGD)血管性痴呆细胞模型细胞黏附分子(CaM)、钙调蛋白激酶Ⅱ(CaMKⅡ)、脑源性神经营养因子(BDNF)及Akt表达的影响。方法选取生长良好的PC12细胞传代、分化,行OGD准备血管性痴...目的研究葛根素对氧糖剥夺(oxygen and glucose deprivation,OGD)血管性痴呆细胞模型细胞黏附分子(CaM)、钙调蛋白激酶Ⅱ(CaMKⅡ)、脑源性神经营养因子(BDNF)及Akt表达的影响。方法选取生长良好的PC12细胞传代、分化,行OGD准备血管性痴呆细胞模型,随机分为对照组、模型组及低、中、高剂量葛根素组。MTT法测定细胞存活率并确定合适的葛根素干预浓度及OGD处理时间;检测乳酸脱氢酶(LDH)释放量评定细胞损伤程度,鉴定细胞模型;Western blot检测CaM、CaMKⅡ、MECP2、BDNF及Akt蛋白的表达水平。结果 PC12细胞存活率随OGD时间延长而逐渐降低,呈时间依赖性;PC12细胞存活率随葛根素浓度增加而逐渐升高,呈浓度依赖性。葛根素有效干预浓度为0.1～10μmol/L;OGD最佳处理时间为6h。与对照组相比,模型组LDH释放量明显增高(P<0.05);葛根素干预组LDH释放量随葛根素浓度增加而减少(P<0.05)。模型组CaM蛋白表达明显升高,BDNF表达量明显减少(P<0.05),MECP2表达及CaMKⅡ、Akt蛋白磷酸化水平均未见明显变化(P>0.05)。葛根素干预可下调CaM蛋白水平,提高MECP2、BDNF的表达及CaMKⅡ磷酸化水平,中、高剂量葛根素组亦能升高Akt蛋白磷酸化水平(P<0.05)。结论葛根素可能通过提高Ca2+-CaM复合物介导CaMKⅡ自身磷酸化水平,诱导MECP2磷酸化,上调BDNF的表达,激活下游PI3K-Akt通路,抑制凋亡基因及蛋白表达,发挥神经保护作用。展开更多
目的观察脂质体介导的VEGF质粒对成年大鼠脑缺血再灌注后大脑神经元phosphorylation of methyl-CpG binding protein 2 (P-MeCP2)表达的影响,探讨VEGF促进缺血损伤后神经元新生的可能机制,进一步为VEGF治疗缺血性脑中风提供实验和理论...目的观察脂质体介导的VEGF质粒对成年大鼠脑缺血再灌注后大脑神经元phosphorylation of methyl-CpG binding protein 2 (P-MeCP2)表达的影响,探讨VEGF促进缺血损伤后神经元新生的可能机制,进一步为VEGF治疗缺血性脑中风提供实验和理论依据。方法 30只SD大鼠,随机分为假手术组(sham组)、质粒组(VEGF组)和对照质粒组(vehicle组),采用左侧大脑中动脉线栓(MCAO)模型,侧脑室给药,免疫印迹、免疫荧光三标染色及激光共聚焦扫描技术等方法检测P-MeCP2的表达。结果再灌2周VEGF质粒组P-MeCP2、BrdU和NeuN同时表达于缺血侧大脑皮质神经元,而缺血对侧没有找到,VEGF质粒组皮质内P-MeCP2的表达较对照组、假手术组显著增高(p<0.01)。结论脂质体介导的VEGF质粒能促进成年大鼠脑缺血神经元的新生,可能与上调P-MeCP2的表达有关。展开更多
Abnormal expression and dysfunction of methyl-CpG binding protein 2 （MeCP2） cause Rett syndrome （RTT）. The diverse phosphorylation modifications modulate MeCP2 function in neural cells. Using western blot and immu...Abnormal expression and dysfunction of methyl-CpG binding protein 2 （MeCP2） cause Rett syndrome （RTT）. The diverse phosphorylation modifications modulate MeCP2 function in neural cells. Using western blot and immunohistochemistry, we examined the expression patterns of MeCP2 and three phospho-MeCP2s （pMeCP2s） in the developing rat brain. The expression of MeCP2 and phospho-S80 （pS80） MeCP2 increased while pS421 MeCP2 and pS292 MeCP2 decreased with brain maturation. In contrast to the nuclear localization of MeCP2 and pS80 MeCP2, pS421 MeCP2 and pS292 MeCP2 were mainly expressed in the cytoplasmic com- partment. Apart from their distribution in neurons, they were also detected at a low level in astrocytes. Postnatallyinitiated MeCP2 deficiency affected cerebellar neural cell development, as determined by the abnormal expression of GFAP, DCX, Tuj 1, MAP-2, and calbindin-D28k. Together, these results demonstrate that MeCP2 and diverse pMeCP2s have distinct features of spatio-temporal expression in the rat brain, and that the precise levels of MeCP2 in the postnatal period are vital to cerebellar neural cell development.展开更多
文摘目的观察脂质体介导的VEGF质粒对成年大鼠脑缺血再灌注后大脑神经元phosphorylation of methyl-CpG binding protein 2 (P-MeCP2)表达的影响,探讨VEGF促进缺血损伤后神经元新生的可能机制,进一步为VEGF治疗缺血性脑中风提供实验和理论依据。方法 30只SD大鼠,随机分为假手术组(sham组)、质粒组(VEGF组)和对照质粒组(vehicle组),采用左侧大脑中动脉线栓(MCAO)模型,侧脑室给药,免疫印迹、免疫荧光三标染色及激光共聚焦扫描技术等方法检测P-MeCP2的表达。结果再灌2周VEGF质粒组P-MeCP2、BrdU和NeuN同时表达于缺血侧大脑皮质神经元,而缺血对侧没有找到,VEGF质粒组皮质内P-MeCP2的表达较对照组、假手术组显著增高(p<0.01)。结论脂质体介导的VEGF质粒能促进成年大鼠脑缺血神经元的新生,可能与上调P-MeCP2的表达有关。
基金supported by a grant from the National Natural Science Foundation of China(81030020)
文摘Abnormal expression and dysfunction of methyl-CpG binding protein 2 （MeCP2） cause Rett syndrome （RTT）. The diverse phosphorylation modifications modulate MeCP2 function in neural cells. Using western blot and immunohistochemistry, we examined the expression patterns of MeCP2 and three phospho-MeCP2s （pMeCP2s） in the developing rat brain. The expression of MeCP2 and phospho-S80 （pS80） MeCP2 increased while pS421 MeCP2 and pS292 MeCP2 decreased with brain maturation. In contrast to the nuclear localization of MeCP2 and pS80 MeCP2, pS421 MeCP2 and pS292 MeCP2 were mainly expressed in the cytoplasmic com- partment. Apart from their distribution in neurons, they were also detected at a low level in astrocytes. Postnatallyinitiated MeCP2 deficiency affected cerebellar neural cell development, as determined by the abnormal expression of GFAP, DCX, Tuj 1, MAP-2, and calbindin-D28k. Together, these results demonstrate that MeCP2 and diverse pMeCP2s have distinct features of spatio-temporal expression in the rat brain, and that the precise levels of MeCP2 in the postnatal period are vital to cerebellar neural cell development.
