In order to investigate the anti-tumor angiogenesis activity with a recombinant Ag43/FGFR1 chimeric protein(AF)vaccine in a mouse H22 hepatoma model,tumor volume and survival rate of the mice were studied at a 3-day i...In order to investigate the anti-tumor angiogenesis activity with a recombinant Ag43/FGFR1 chimeric protein(AF)vaccine in a mouse H22 hepatoma model,tumor volume and survival rate of the mice were studied at a 3-day interval.Microvessel density(MVD)was detected by immunohistochemistry.The endothelial deposition of autoantibodies within tumor tissues was examined by immunofluorescent staining,and anti-FGFR1 antibody-producing B cells(APBCs)were tested by enzyme-linked immunospot(ELISPOT)assay.Compared with t...展开更多
目的观察葡萄籽原花青素(GSPE)对放射性脑损伤大鼠的防治作用,并分析可能的保护机制。方法 72只3月龄雄性Sprague-Dawley大鼠随机分为对照组(n=18)、模型组(n=18)、低剂量GSPE组(n=18)和高剂量GSPE组(n=18)。用直线加速器进行脑部照射22...目的观察葡萄籽原花青素(GSPE)对放射性脑损伤大鼠的防治作用,并分析可能的保护机制。方法 72只3月龄雄性Sprague-Dawley大鼠随机分为对照组(n=18)、模型组(n=18)、低剂量GSPE组(n=18)和高剂量GSPE组(n=18)。用直线加速器进行脑部照射22 Gy制作放射性脑损伤模型。采用穿梭箱实验检测大鼠学习能力;HE染色观察海马区神经细胞组织形态变化;RT-PCR法检测生长相关蛋白-43(GAP-43)m RNA表达,Western blotting检测磷酸化的细胞外信号调节激酶1/2(ERK1/2)表达。结果与模型组比较,各GSPE组动物穿梭箱主动回避反应率显著升高(P<0.001),被动回避潜伏期显著缩短(P<0.001);海马区神经细胞结构损伤减轻;GAP-43 m RNA和磷酸化ERK1/2表达水平增高(P<0.001)。且高剂量GSPE组显著优于低剂量GSPE组(P<0.001)。模型组中GAP-43 m RNA表达水平与磷酸化ERK1/2水平呈正相关(r=0.764,P<0.001);低剂量GSPE组和高剂量GSPE组中GAP-43 m RNA表达水平与磷酸化ERK1/2水平呈正相关(r=0.814,0.822,P<0.001)。结论 GSPE通过ERK1/2途径提高大鼠海马区GAP-43表达,发挥防治放射性脑损伤作用。展开更多
OBJECTIVE Astrocytic gap junctions formed by connexin 43(Cx43) are crucial for intercellular communication between spinal cord astrocytes. Various neurological disorders include chronic pain are associated with dysfun...OBJECTIVE Astrocytic gap junctions formed by connexin 43(Cx43) are crucial for intercellular communication between spinal cord astrocytes. Various neurological disorders include chronic pain are associated with dysfunctional Cx43-gap junctions. A previous study showed that treatment of spinal astrocytes in culture with pro-inflammatory cytokines tumor necrosis factor-α(TNF-α) and interferon-γ(IFN-γ) decreased both Cx43 expression and gap junction intercellular communication(GJIC)via a c-jun terminal kinase-dependent pathway. However,the exact mechanism that Cx43 does this in the context of spinal astrocytic dysfunction is unclear. The current study investigated the downstream signaling of Cx43-gap junction in rat primary cultured spinal astrocytes stimulated with cytokines.METHODS Wefocused on the glutamate transporters,examined the alteration of GLT-1 and glutamate-aspartate transporter(GLAST)expression and function in rat primary cultured spinal astrocytes stimulated with cytokines by real-time PCR and Western blotting. The function of GLT-1 was analyzed using the carbon 14. To elucidate the effect of Cx43 on glutamate transporters in spinal astrocytes,changes in glutamate transporters expression and function were quantified after Cx43 siRNA treatment.RESULTS The transcriptional and translational levels of Cx43 were reduced after 12 hr co-treatment with TNF-α(10 ng·mL^(-1)) or IFN-γ(5 ng·mL^(-1)). Furthermore,transcriptional and translational levels of GLT-1 and GLAST were also significantly reduced 24 and 48 h co-treatment with TNF-α or IFN-γ.Moreover,functional GLT-1 and GLAST uptake were inhibited by the mixture of TNF-α and IFN-γ. In addition,Both the decrease of GLT-1 expression and the reduction in functional GLT-1 uptake induced by the Cx43 si RNA,but both the expression and functional GLAST were no changes. CONCLUSION These results indicate that a Cx43 downregulation is induced under inflammatory condition that disrupts spinal astrocytic GLT-1 expression and function,leading to astrocytic dysfunction and the maintenance of the neuroinflammatory state.展开更多
基金supported by the National Natural Sciences Foundation of China(No.3066005530670548+3 种基金30900663)Program for New Century Excellent Talents in University of China(NCET-08-657)the Key Project of Chinese Ministry of Education(No.209097)a grant from Hainan Provincial Natural Sciences Foundation(No.30864)
文摘In order to investigate the anti-tumor angiogenesis activity with a recombinant Ag43/FGFR1 chimeric protein(AF)vaccine in a mouse H22 hepatoma model,tumor volume and survival rate of the mice were studied at a 3-day interval.Microvessel density(MVD)was detected by immunohistochemistry.The endothelial deposition of autoantibodies within tumor tissues was examined by immunofluorescent staining,and anti-FGFR1 antibody-producing B cells(APBCs)were tested by enzyme-linked immunospot(ELISPOT)assay.Compared with t...
文摘目的观察葡萄籽原花青素(GSPE)对放射性脑损伤大鼠的防治作用,并分析可能的保护机制。方法 72只3月龄雄性Sprague-Dawley大鼠随机分为对照组(n=18)、模型组(n=18)、低剂量GSPE组(n=18)和高剂量GSPE组(n=18)。用直线加速器进行脑部照射22 Gy制作放射性脑损伤模型。采用穿梭箱实验检测大鼠学习能力;HE染色观察海马区神经细胞组织形态变化;RT-PCR法检测生长相关蛋白-43(GAP-43)m RNA表达,Western blotting检测磷酸化的细胞外信号调节激酶1/2(ERK1/2)表达。结果与模型组比较,各GSPE组动物穿梭箱主动回避反应率显著升高(P<0.001),被动回避潜伏期显著缩短(P<0.001);海马区神经细胞结构损伤减轻;GAP-43 m RNA和磷酸化ERK1/2表达水平增高(P<0.001)。且高剂量GSPE组显著优于低剂量GSPE组(P<0.001)。模型组中GAP-43 m RNA表达水平与磷酸化ERK1/2水平呈正相关(r=0.764,P<0.001);低剂量GSPE组和高剂量GSPE组中GAP-43 m RNA表达水平与磷酸化ERK1/2水平呈正相关(r=0.814,0.822,P<0.001)。结论 GSPE通过ERK1/2途径提高大鼠海马区GAP-43表达,发挥防治放射性脑损伤作用。
文摘目的观察微小RNA-1(miRNA-1)对自发性高血压大鼠(SHRs)肥厚左室心肌组织中连接蛋白43(Cx43)表达的调控及其意义。方法 18只17周龄雄性SHRs随机分为干预组(n=6)、阴性对照组(n=6)和空白对照组(n=6),通过脂质体瞬时转染技术,干预组由尾静脉注射miRNA-1抑制剂,两对照组分别注入miRNA-1抑制剂阴性对照和无血清培养基混合物,并通过实时荧光定量PCR、免疫组织化学法及western blot等技术,检测大鼠左室心肌组织miRNA-1及Cx43蛋白表达水平的改变。结果与阴性对照组和空白对照组比较,干预组miRNA-1表达水平明显降低,伴随着Cx43蛋白表达水平增高(0.45±0.15 vs 0.27±0.14,0.23±0.10,P均<0.05)。结论抑制miRNA-1表达能使SHR肥厚左室心肌组织Cx43蛋白水平升高。
文摘OBJECTIVE Astrocytic gap junctions formed by connexin 43(Cx43) are crucial for intercellular communication between spinal cord astrocytes. Various neurological disorders include chronic pain are associated with dysfunctional Cx43-gap junctions. A previous study showed that treatment of spinal astrocytes in culture with pro-inflammatory cytokines tumor necrosis factor-α(TNF-α) and interferon-γ(IFN-γ) decreased both Cx43 expression and gap junction intercellular communication(GJIC)via a c-jun terminal kinase-dependent pathway. However,the exact mechanism that Cx43 does this in the context of spinal astrocytic dysfunction is unclear. The current study investigated the downstream signaling of Cx43-gap junction in rat primary cultured spinal astrocytes stimulated with cytokines.METHODS Wefocused on the glutamate transporters,examined the alteration of GLT-1 and glutamate-aspartate transporter(GLAST)expression and function in rat primary cultured spinal astrocytes stimulated with cytokines by real-time PCR and Western blotting. The function of GLT-1 was analyzed using the carbon 14. To elucidate the effect of Cx43 on glutamate transporters in spinal astrocytes,changes in glutamate transporters expression and function were quantified after Cx43 siRNA treatment.RESULTS The transcriptional and translational levels of Cx43 were reduced after 12 hr co-treatment with TNF-α(10 ng·mL^(-1)) or IFN-γ(5 ng·mL^(-1)). Furthermore,transcriptional and translational levels of GLT-1 and GLAST were also significantly reduced 24 and 48 h co-treatment with TNF-α or IFN-γ.Moreover,functional GLT-1 and GLAST uptake were inhibited by the mixture of TNF-α and IFN-γ. In addition,Both the decrease of GLT-1 expression and the reduction in functional GLT-1 uptake induced by the Cx43 si RNA,but both the expression and functional GLAST were no changes. CONCLUSION These results indicate that a Cx43 downregulation is induced under inflammatory condition that disrupts spinal astrocytic GLT-1 expression and function,leading to astrocytic dysfunction and the maintenance of the neuroinflammatory state.
