目的探讨β1-肾上腺素能受体阻滞剂艾司洛尔在大约克猪室颤心脏停搏心肺复苏模型的作用以及潜在的机制。方法50只健康成年雄性大约克猪在室颤4min后根据药物干预随机分为假手术组（Sham组）、肾上腺素组（AD组）、肾上腺素和艾司洛尔联...目的探讨β1-肾上腺素能受体阻滞剂艾司洛尔在大约克猪室颤心脏停搏心肺复苏模型的作用以及潜在的机制。方法50只健康成年雄性大约克猪在室颤4min后根据药物干预随机分为假手术组（Sham组）、肾上腺素组（AD组）、肾上腺素和艾司洛尔联合用药组（AD＋ES组），监测复苏后72h心电图，比较各组的心率、血压、冠脉灌注压、自主循环恢复、心功能等指标。死亡动物迅速取材，应用实时荧光定量PCR技术、免疫印迹技术检测左室基底部外膜层心肌钙／钙调素依赖性蛋白激酶Ⅱδ（calcium／calmodulin dependent protein kinaseⅡδ，CaMKⅡδ）的mRNA转录水平和蛋白表达水平。结果两组均有一只动物未能成功复苏。在复苏过程中两组自主循环恢复率、冠脉灌注压的差异无统计学意义。与Sham组相比，AD组和AD＋ES组复苏后心功能下降，但自主循环恢复后2h，AD＋ES组射血分数和心输出量显著高于AD组，更接近于Sham组。AD＋ES组复苏后72h恶性心律失常的发生率少于AD组，短期生存率高于AD组，心肌CaMKⅡδmRNA的相对表达量显著低于AD组[（1．76±0．12）雠（1．22±0．10），t=15．07，P=0．0001]。两组复苏72h内心源性死亡动物心肌CaMKⅡδ的mRNA相对表达量〉非心源性死亡动物〉72h后存活动物。蛋白免疫印迹结果表明各组CaMKⅡδ蛋白表达水平的差异与mRNA表达水平的差异是一致的。结论肾上腺素和艾司洛尔联合应用改善心肺复苏后心功能，减少恶性心律失常的发生，提高复苏后72h生存率。下调CaMKⅡδ的mRNA和蛋白表达水平是艾司洛尔减少复苏后室性心律失常发生的潜在分子学基础机制。展开更多
Background The effect of chronic stress on cognitive functions has been one of the hot topic in neuroscience. But there has been much controversy over its mechanism. Such single stressor applied in the past could not ...Background The effect of chronic stress on cognitive functions has been one of the hot topic in neuroscience. But there has been much controversy over its mechanism. Such single stressor applied in the past could not simulate complicated living circumstances that people confronted with. The aim of this study was to investigate the effects of chronic multiple-stress on learning and memory as well as on the levels of calcium/calmodulin-dependent protein kinase Ⅱ（CaMKⅡ）, calmodulin （CAM） mRNA, and cAMP-response element binding protein （CREB） mRNA in the hippocampus of rats. Methods The rats were divided randomly into stressed and control groups. The stressed group was given chronic multiple-stress for 6 weeks to set up a chronic multiple-stressed model. The rats＇ performance of spatial learning and memory was tested using Morris Water Maze （MWM） and Y-maze. Meanwhile, the expressions of CAMKII, CAM mRNA and CREB mRNA of rats＇ hippocampus were detected by immunohistochemistry, Western blot and reverse transcription-polymerase chain reaction （RT-PCR）, respectively. In addition, the width of synaptic cleft and the thickness of post-synaptic densities （PSD） were observed in the hippocampal CA3 region of rats by electron microscopy. Results After exposure to chronic multiple-stress for 6 weeks, the ability of learning and memory of the stressed group was higher than that of the control group （P〈0.05, P〈0.01）. The width of synaptic cleft was smaller and the thickness of PSD was larger in the hippocampal CA3 region of the stressed group than in that of the control group （P〈0.01）. The CAMKII immunostaining of the stressed group was stronger than that of the control group in the stratum radiatum and oriens of the hippocampal CA1 and CA3, especially in the stratum oriens. Quantitative analysis indicated that the expression of CAMKII, CAM mRNA, and CREB mRNA in the hippocampus of the stressed group was higher than that of the control group （P〈0.05, P〈0.01）. Conclusions The 展开更多
钙/钙调蛋白依赖性蛋白激酶Ⅱ(calcium/calmodulin-dependent protein kinaseⅡ,CaMKⅡ)是一种重要的丝氨酸/苏氨酸蛋白激酶,参与脑内多种神经活动的调节,并被认为是一个"记忆分子",在突触可塑性和学习记忆中发挥关键作用。C...钙/钙调蛋白依赖性蛋白激酶Ⅱ(calcium/calmodulin-dependent protein kinaseⅡ,CaMKⅡ)是一种重要的丝氨酸/苏氨酸蛋白激酶,参与脑内多种神经活动的调节,并被认为是一个"记忆分子",在突触可塑性和学习记忆中发挥关键作用。CaMKⅡ的功能正常对于大脑的正常生理活动是必需的。近年来研究发现,许多神经精神疾病的发生都与脑内CaMKⅡ的功能异常有关。该文对CaMKⅡ在突触可塑性以及神经精神疾病中的作用做一综述。展开更多
钙调素依赖性蛋白激酶Ⅱ(calcium/calmodulin-dependent protein kinaseⅡ,CaMKⅡ)是一种在学习和记忆形成机制中具有重要作用的蛋白激酶,它存在于大多数组织中,但以神经元中表达量最高。大多数具有激酶活性的蛋白分子在组织中的表达量...钙调素依赖性蛋白激酶Ⅱ(calcium/calmodulin-dependent protein kinaseⅡ,CaMKⅡ)是一种在学习和记忆形成机制中具有重要作用的蛋白激酶,它存在于大多数组织中,但以神经元中表达量最高。大多数具有激酶活性的蛋白分子在组织中的表达量都相对较低,所以CaMKⅡ在神经系统中高度表达具有其特殊意义。该文将较为全面的对CaMKⅡ的分子结构与自身磷酸化特征、亚细胞空间定位及其在突触可塑性中的作用进行综述。展开更多
目的急性缺血缺氧可致心功能受损和心律失常,钙离子在其中起重要作用,慢性缺氧对心功能及心肌细胞内钙离子活动同样产生影响,但机制不同。该研究拟通过慢性缺氧动物模型,研究慢性缺氧对心肌细胞内钙调素(calmodulin,CaM)、钙/钙调素依...目的急性缺血缺氧可致心功能受损和心律失常,钙离子在其中起重要作用,慢性缺氧对心功能及心肌细胞内钙离子活动同样产生影响,但机制不同。该研究拟通过慢性缺氧动物模型,研究慢性缺氧对心肌细胞内钙调素(calmodulin,CaM)、钙/钙调素依赖性蛋白激酶Ⅱ(calcium/calmodulin-dependent protein kinase Ⅱ,CaMKⅡ)的表达及其对细胞内Ca2+活动的影响,深入了解慢性缺氧对心脏功能及电活动的影响机制。方法通过吸入低浓度含氧气体(FiO2:10%)建立慢性缺氧大鼠动物模型。在实验1周和3周时,应用RT-PCR,Western Blot方法分别检测对照组和慢性缺氧组动物心肌细胞内CaM和CaMKⅡγ、CaMKIIδmRNA和蛋白表达;分离并培养正常心肌细胞和缺氧3周细胞,应用激光共聚焦法分别检测两种心肌细胞内Ca2+活动,同时应用CaMKⅡ特异性抑制剂KN-62,观察CaMKII在慢性缺氧下对心肌细胞内Ca2+活动的影响。结果实验1周和3周时,慢性缺氧大鼠心肌细胞内CaM和CaMKIIγ、CaMKIIδ的mRNA和蛋白表达均较正常动物高(P<0.01);在缺氧1周和3周组动物间CaM和CaMKIIδ也存在差异(P<0.01),但CaMKⅡγ无差异(P>0.05)。激光共聚焦研究发现,慢性缺氧心肌细胞内Ca2+活动其钙波振幅虽和正常心肌细胞无差异(P>0.05),但钙波时程延长(P<0.01);应用KN-62后,慢性缺氧动物钙波振幅和时程改变较明显(P<0.01)。结论慢性缺氧可使大鼠心肌细胞内CaM和CaMKⅡ合成代偿性增加,保持大鼠心肌细胞内钙稳态,从而在一定时期内维持心功能稳定。但随缺氧时间延长,心功能可受损并可致心律失常。展开更多
In the present study, we examined the analgesic effect of repeated electroacupuncture at bilateral Zusanfi （ST36） and Yanglingquan （GB34） once a day for 14 consecutive days in a rat model of chronic sciatic nerve ...In the present study, we examined the analgesic effect of repeated electroacupuncture at bilateral Zusanfi （ST36） and Yanglingquan （GB34） once a day for 14 consecutive days in a rat model of chronic sciatic nerve constriction injury-induced neuropathic pain. In addition, concomitant changes in calcium/calmodulin-dependent protein kinase II expression and synaptic ultrastructure of neurons in the hippocampal CA3 region were examined. The thermal pain threshold （paw withdrawal latency） was increased significantly in both groups at 2 weeks after electroacupuncture intervention compared with 2 days of electroacupuncture. In ovariectomized rats with chronic constriction injury, the analgesic effect was significantly reduced. Electroacupuncture for 2 weeks significantly diminished the injury-induced increase in synaptJc cleft width and thinning of the postsynaptJc density, and it significantly suppressed the down-regulation of intracellular calcium/ calmodulin-dependent protein kinase II expression in the hippocampal CA3 region. Repeated electroacupuncture intervention had a cumulative analgesic effect on injury-induced neuropathic pain reactions, and it led to synaptic remodeling of hippocampal neurons and upregulated calcium/calmodulin-dependent protein kJnase II expression in the hippocampal CA3 region.展开更多
文摘目的探讨β1-肾上腺素能受体阻滞剂艾司洛尔在大约克猪室颤心脏停搏心肺复苏模型的作用以及潜在的机制。方法50只健康成年雄性大约克猪在室颤4min后根据药物干预随机分为假手术组（Sham组）、肾上腺素组（AD组）、肾上腺素和艾司洛尔联合用药组（AD＋ES组），监测复苏后72h心电图，比较各组的心率、血压、冠脉灌注压、自主循环恢复、心功能等指标。死亡动物迅速取材，应用实时荧光定量PCR技术、免疫印迹技术检测左室基底部外膜层心肌钙／钙调素依赖性蛋白激酶Ⅱδ（calcium／calmodulin dependent protein kinaseⅡδ，CaMKⅡδ）的mRNA转录水平和蛋白表达水平。结果两组均有一只动物未能成功复苏。在复苏过程中两组自主循环恢复率、冠脉灌注压的差异无统计学意义。与Sham组相比，AD组和AD＋ES组复苏后心功能下降，但自主循环恢复后2h，AD＋ES组射血分数和心输出量显著高于AD组，更接近于Sham组。AD＋ES组复苏后72h恶性心律失常的发生率少于AD组，短期生存率高于AD组，心肌CaMKⅡδmRNA的相对表达量显著低于AD组[（1．76±0．12）雠（1．22±0．10），t=15．07，P=0．0001]。两组复苏72h内心源性死亡动物心肌CaMKⅡδ的mRNA相对表达量〉非心源性死亡动物〉72h后存活动物。蛋白免疫印迹结果表明各组CaMKⅡδ蛋白表达水平的差异与mRNA表达水平的差异是一致的。结论肾上腺素和艾司洛尔联合应用改善心肺复苏后心功能，减少恶性心律失常的发生，提高复苏后72h生存率。下调CaMKⅡδ的mRNA和蛋白表达水平是艾司洛尔减少复苏后室性心律失常发生的潜在分子学基础机制。
基金supported by High School Science and Technology Fund Planning Project of Tianjin Municipality,China(No.20060206)the Scientific Research Fund of Tianjin Medical University,China(No.2011ky33)
基金This study was supported by a grant from the National Natural Science Foundation of China (No. 30270446).
文摘Background The effect of chronic stress on cognitive functions has been one of the hot topic in neuroscience. But there has been much controversy over its mechanism. Such single stressor applied in the past could not simulate complicated living circumstances that people confronted with. The aim of this study was to investigate the effects of chronic multiple-stress on learning and memory as well as on the levels of calcium/calmodulin-dependent protein kinase Ⅱ（CaMKⅡ）, calmodulin （CAM） mRNA, and cAMP-response element binding protein （CREB） mRNA in the hippocampus of rats. Methods The rats were divided randomly into stressed and control groups. The stressed group was given chronic multiple-stress for 6 weeks to set up a chronic multiple-stressed model. The rats＇ performance of spatial learning and memory was tested using Morris Water Maze （MWM） and Y-maze. Meanwhile, the expressions of CAMKII, CAM mRNA and CREB mRNA of rats＇ hippocampus were detected by immunohistochemistry, Western blot and reverse transcription-polymerase chain reaction （RT-PCR）, respectively. In addition, the width of synaptic cleft and the thickness of post-synaptic densities （PSD） were observed in the hippocampal CA3 region of rats by electron microscopy. Results After exposure to chronic multiple-stress for 6 weeks, the ability of learning and memory of the stressed group was higher than that of the control group （P〈0.05, P〈0.01）. The width of synaptic cleft was smaller and the thickness of PSD was larger in the hippocampal CA3 region of the stressed group than in that of the control group （P〈0.01）. The CAMKII immunostaining of the stressed group was stronger than that of the control group in the stratum radiatum and oriens of the hippocampal CA1 and CA3, especially in the stratum oriens. Quantitative analysis indicated that the expression of CAMKII, CAM mRNA, and CREB mRNA in the hippocampus of the stressed group was higher than that of the control group （P〈0.05, P〈0.01）. Conclusions The
文摘钙/钙调蛋白依赖性蛋白激酶Ⅱ(calcium/calmodulin-dependent protein kinaseⅡ,CaMKⅡ)是一种重要的丝氨酸/苏氨酸蛋白激酶,参与脑内多种神经活动的调节,并被认为是一个"记忆分子",在突触可塑性和学习记忆中发挥关键作用。CaMKⅡ的功能正常对于大脑的正常生理活动是必需的。近年来研究发现,许多神经精神疾病的发生都与脑内CaMKⅡ的功能异常有关。该文对CaMKⅡ在突触可塑性以及神经精神疾病中的作用做一综述。
文摘钙调素依赖性蛋白激酶Ⅱ(calcium/calmodulin-dependent protein kinaseⅡ,CaMKⅡ)是一种在学习和记忆形成机制中具有重要作用的蛋白激酶,它存在于大多数组织中,但以神经元中表达量最高。大多数具有激酶活性的蛋白分子在组织中的表达量都相对较低,所以CaMKⅡ在神经系统中高度表达具有其特殊意义。该文将较为全面的对CaMKⅡ的分子结构与自身磷酸化特征、亚细胞空间定位及其在突触可塑性中的作用进行综述。
文摘目的急性缺血缺氧可致心功能受损和心律失常,钙离子在其中起重要作用,慢性缺氧对心功能及心肌细胞内钙离子活动同样产生影响,但机制不同。该研究拟通过慢性缺氧动物模型,研究慢性缺氧对心肌细胞内钙调素(calmodulin,CaM)、钙/钙调素依赖性蛋白激酶Ⅱ(calcium/calmodulin-dependent protein kinase Ⅱ,CaMKⅡ)的表达及其对细胞内Ca2+活动的影响,深入了解慢性缺氧对心脏功能及电活动的影响机制。方法通过吸入低浓度含氧气体(FiO2:10%)建立慢性缺氧大鼠动物模型。在实验1周和3周时,应用RT-PCR,Western Blot方法分别检测对照组和慢性缺氧组动物心肌细胞内CaM和CaMKⅡγ、CaMKIIδmRNA和蛋白表达;分离并培养正常心肌细胞和缺氧3周细胞,应用激光共聚焦法分别检测两种心肌细胞内Ca2+活动,同时应用CaMKⅡ特异性抑制剂KN-62,观察CaMKII在慢性缺氧下对心肌细胞内Ca2+活动的影响。结果实验1周和3周时,慢性缺氧大鼠心肌细胞内CaM和CaMKIIγ、CaMKIIδ的mRNA和蛋白表达均较正常动物高(P<0.01);在缺氧1周和3周组动物间CaM和CaMKIIδ也存在差异(P<0.01),但CaMKⅡγ无差异(P>0.05)。激光共聚焦研究发现,慢性缺氧心肌细胞内Ca2+活动其钙波振幅虽和正常心肌细胞无差异(P>0.05),但钙波时程延长(P<0.01);应用KN-62后,慢性缺氧动物钙波振幅和时程改变较明显(P<0.01)。结论慢性缺氧可使大鼠心肌细胞内CaM和CaMKⅡ合成代偿性增加,保持大鼠心肌细胞内钙稳态,从而在一定时期内维持心功能稳定。但随缺氧时间延长,心功能可受损并可致心律失常。
基金supported by the National Natural Science Foundation of China,No.30472241,90709031 and 30973796the Ministry of Science and Technology of China("973"Project),No.2007CB512505+1 种基金provided by the Foundation of Hainan Province,No.310054the Health Department of Hainan Province,Qiong-Wei-45
文摘In the present study, we examined the analgesic effect of repeated electroacupuncture at bilateral Zusanfi （ST36） and Yanglingquan （GB34） once a day for 14 consecutive days in a rat model of chronic sciatic nerve constriction injury-induced neuropathic pain. In addition, concomitant changes in calcium/calmodulin-dependent protein kinase II expression and synaptic ultrastructure of neurons in the hippocampal CA3 region were examined. The thermal pain threshold （paw withdrawal latency） was increased significantly in both groups at 2 weeks after electroacupuncture intervention compared with 2 days of electroacupuncture. In ovariectomized rats with chronic constriction injury, the analgesic effect was significantly reduced. Electroacupuncture for 2 weeks significantly diminished the injury-induced increase in synaptJc cleft width and thinning of the postsynaptJc density, and it significantly suppressed the down-regulation of intracellular calcium/ calmodulin-dependent protein kinase II expression in the hippocampal CA3 region. Repeated electroacupuncture intervention had a cumulative analgesic effect on injury-induced neuropathic pain reactions, and it led to synaptic remodeling of hippocampal neurons and upregulated calcium/calmodulin-dependent protein kJnase II expression in the hippocampal CA3 region.
