Effects of RSM and astragus on expression of Fos and Jun proteins in rat brains after cerebral ischemia and reperfusion

AIM: To investigate the expression of Fos and Jun proteins in rat brains after focal cerebral ischemia followed by reperfusion and effects of RSM and astragus.METHODS: 30 SD adult male rats were divided into 5 groups at random .Group A:sham operated group;Group B:model group;Group C:treated with RSM;Group D:treated with astragus;Group E:treated with RSM and astragus.The immunohistochemistry and medical image processing system(MIPS)were used to measure the numbers and mean grey levels of Fos and Jun protein positive cells in rat cerebral cortex of 5 groups. RESULTS: (1) In cerebral cortex of group B ,C ,D ,E,the numbers of Fos and Jun positive cells were more than those in group A and mean grey levels of Fos and Jun positive cells were lower than those in group A(P< 0.01);(2) In cerebral cortex of ischemic sides in group C,D,E,the numbers of Fos and Jun positive cells were less than those in group B and mean grey levels of Fos and Jun positive cells were higher than those in group B(P< 0.01) ;(3) Group E had more significant effects than group C or group D (P< 0.01). CONCLUSION: The expression of Fos protein and Jun protein in model group increased significantly,compared with sham operated group;RSM ,astragus ,RSM and astragus all could inhibit partly the expression of Fos protein and Jun protein after cerebral ischemia and reperfusion;Prescription of RSM and astragus had stronger inhibiting effects than RSM or astragus.It may be one of mechanisms that ischemic stoke is treated by reinforcing Qi and activating blood circulation therapy in TCM clinic.

Optimal compatible doses and effects of ephedrine and naloxone on neural plasticity in cerebral ischemia/reperfusion rats

BACKGROUND： Ephedrine promotes neural plasticity in rats following cerebral ischemia/reperfusion injury. Ephedrine has been combined with naloxone in some studies, and it has been confirmed that their combination has synergistic effects on increasing neural plasticity following cerebral ischemia/reperfusion injury. OBJECTIVE： To investigate the effects of ephedrine combined with various doses of naloxone on neural plasticity and to find an optimal dose of naloxone in rats after cerebral ischemia/reperfusion injury by analyzing growth associated protein-43 （GAP-43）, synaptophysin and β-endorphin expression in the hippocampal CA3 area. DESIGN, TIME AND SETTING： This immunohistochemical, randomized, controlled, animal experiment was performed at the Chongqing Research Institute of Pediatrics, China from September 2007 to June 2008. MATERIALS： Ephedrine hydrochloride injection and naloxone hydrochloride injection were respectively purchased from Shandong Lvliang Pharmaceutical Factory, China and Sichuan Jingwei Pharmaceutical Co., Ltd., China. A total of 192 healthy adult Sprague Dawley rats were used to establish models of left middle cerebral artery occlusion using the suture occlusion method. METHODS： At 2 hours following cerebral ischemia, the rats were intraperitoneally injected with 1.5 mg/kg/d ephedrine （ephedrine group）, with 0.1, 0.2, or 0.3 mg/kg/d naloxone （low, moderate and high doses of naloxone groups）, with 1.5 mg/kg/d ephedrine ＋ 0.1, 0.2, or 0.3 mg/kg/d naloxone （ephedrine ＋ low, moderate and high doses of naloxone groups）, and with 0.5 mL saline （model group）, respectively. MAIN OUTCOME MEASURES： GAP-43, synaptophysin and β -endorphin expression were detected in the hippocampal CA3 area using immunohistochemistry 1-4 weeks after surgery. Sensorimotor integration in rats was assessed using the beam walking test. RESULTS： GAP-43 and synaptophysin expression was greater in the ephedrine group, and in the ephedrine ＋ moderate and high doses of naloxone groups compared with the model group. GAP-43 and synaptophysin expression was greatest in the ephedrine ＋ high dose of naloxone group at 2 and 3 weeks alter surgery. β -endorphin expression was significantly lower in the ephedrine group, and in the ephedrine ＋ moderate and high doses of naloxone groups compared with the model group （P 〈 0.05）. β -endorphin expression was persistently low in the ephedrine ＋ high dose of naloxone group. At 1-3 weeks after surgery, the beam walking test score was significantly higher in the ephedrine group and ephedrine ＋ various doses of naloxone groups than in the model group （P 〈 0.05）. The score was higher in the ephedrine ＋ moderate and high doses of naloxone groups than in the ephedrine group （P 〈 0.05）. Moreover, the score was increased as the dose of naloxone increased in the ephedrine ＋ various doses of naloxone groups. CONCLUSION： Ephedrine promotes GAP-43 and synaptophysin expression, inhibits /3 -endorphin expression in the hippocampal CA3 area, and improves motor function in rats following cerebral ischemia/reperfusion injury. Naloxone does not have the above-mentioned effects. During combined treatment with ephedrine and naloxone, however, the above-described effects are enhanced with an increased dose of naloxone. The combination of ephedrine （1.5 mg/kg/d） and naloxone （0.3 mg/kg/d） can produce optimal therapeutic efficacy in treatment of cerebral ischemic injury.

大鼠脑缺血再灌注区ICAM-1表达与白细胞浸润的观察及丹参的影响

目的 观察细胞间粘附分子（ Ｉ Ｃ Ａ Ｍ１） 蛋白在大鼠脑缺血再灌注的不同时程脑组织中的表达与中性白细胞浸润程度的关系及丹参对它们的影响。方法 Ｓ Ｄ大鼠分为３ 组：假手术组、对照组及丹参组。大脑中动脉缺血２ ｈ 再灌注２ ｈ 、１２ ｈ、２４ ｈ 、４８ ｈ 、７２ ｈ 、７ ｄ、１４ ｄ 后，分别进行 Ｉ Ｃ Ａ Ｍ１ 免疫组织化学及组织 Ｈ Ｅ染色。结果 在脑缺血再灌早期，脑微血管内皮细胞 Ｉ Ｃ Ａ Ｍ１ 免疫反应开始逐渐增加，再灌注４８ ｈ 达到高峰，再灌注１４ ｄ 接近正常水平，同时脑缺血区中性白细胞浸润也随之增加，在时程上与 Ｉ Ｃ Ａ Ｍ１ 表达同步。丹参组，再灌注４８ ｈ 后， Ｉ Ｃ Ａ Ｍ１ 免疫阳性血管数及中性白细胞的浸润比同时间对照组明显降低。结论 脑缺血 Ｉ Ｃ Ａ Ｍ１ 的表达与中性白细胞浸润密切相关，丹参能降低 Ｉ Ｃ Ａ Ｍ１ 的表达，抑制中性白细胞的浸润。

丹参、黄芪对脑缺血再灌注后BCL-2蛋白表达影响的实验研究

目的 :探讨局灶性脑缺血再灌注后脑组织BCL -2蛋白的表达及丹参、黄芪、丹参与黄芪合用对其表达的影响。方法 :采用免疫组化与医学图像分析检测假手术组、模型组及丹参、黄芪、丹参与黄芪合用组大鼠大脑皮层BCL -2蛋白阳性细胞的数目、平均灰度。结果 :①与假手术组比较 ,模型组脑组织缺血侧BCL -2阳性细胞数目增多 ,平均灰度值降低 (P <0 .0 1 )。②在缺血侧脑组织 ,各药物干预组与模型组比较 ,BCL -2阳性细胞数目增多 ,平均灰度值降低 (P <0 .0 1 ) ,其中丹参与黄芪合用组效果最显著 (P <0 .0 1 )。结论 :局灶性脑缺血再灌注后脑组织BCL -2表达显著升高 ;丹参、黄芪及两药合用均可增强BCL -2表达 ,两药合用比单用丹参或黄芪上调BCL -2表达更显著。这可能是临床应用益气活血治则治疗缺血性中风的机理之一。

缺血性脑卒中与脑组织β—EP含量及脑比重的关系及丹参的影响

在沙土鼠脑缺血模型上，用放射免疫分析法测定了脑组织中的β－内啡肽（β－Ｅｎｄｏｒｐｈｉｎ，β－ＥＰ）的含量，观察了缺血后脑组织β－ＥＰ含量的变化及丹参对缺血后β－ＥＰ含量变化和卒中症候的影响。结果显示：沙土鼠缺血８ｈ后β－ＥＰ含量显著增加，且与卒中定候呈正相关，与脑比重呈负相关。丹参可以改善卒中症候，减轻脑水肿，抑制缺血后β－ＥＰ的升高．提示缺血后中枢β－ＥＰ的增加将会加重缺血性损伤，丹参对内源性阿片肽系统具有抑制作用，这可能是其抗缺血性脑损伤的机理之一。

丹参抗脑缺血在神经细胞、分子及递质水平的作用机制研究进展

目的为丹参抗脑缺血的药理及临床研究提供理论依据。方法查阅1989-2006年清华同方数据库,回顾分析丹参在神经细胞、分子及递质水平的脑缺血保护作用机制的研究报道。结果丹参抗脑缺血在神经细胞、分子及递质水平的作用机制主要有:通过抑制缺血区小胶质细胞的活化、抑制ICAM-1的表达等降低神经细胞的损伤;通过下调ICE表达、上调bcl-2蛋白的表达等抑制神经细胞的凋亡;通过减少NO合成、抑制EAA释放等影响神经递质水平;通过抑制ET-1表达、调节TXA2-PGI2平衡等影响血管相关活性物质的表达。结论丹参可从降低神经细胞的损伤、抑制神经细胞的凋亡、影响神经递质及血管活性物质的表达等多个方面起到脑缺血保护作用。

c-Jun、bFGF、HSP70在丹参改善颞叶缺血大鼠空间记忆障碍中的表达变化

目的：探讨在丹参改善单侧颞叶缺血大鼠空间记忆障碍中c-Jun，bFGF和HSP70的表达变化。方法：采用立体定向光化学方法选择性地导致大鼠左侧颞叶皮层缺血，术前30分钟及术后第3天分别给丹参组大鼠腹腔注射丹参10g／kg，用Morris水迷宫及图像自动监视系统监测大鼠行为。然后取脑进行病理学及c-Jun、bFGF和HSP70免疫组化分析。结果：经丹参治疗，颞叶缺血性损害大鼠的空间记忆障碍得到显著改善，颞叶缺血损害程度显著减轻，缺血灶内c-Jun、bFGF和HSP70表达明显减少。结论：丹参可以明显改善单侧颞叶缺血性损害大鼠的空间记忆障碍，其机制可能与丹参减轻颞叶缺血损害，下调c－Jun、bFGF和HSP70表达有关。

脑缺血再灌注脑内纹状体区细胞外液谷胱甘肽的动态变化及丹参的影响

采用脑内微透析技术，应用高压液相色谱电化学检测方法（ＨＰＬＣ－ＥＤ），活体动态观察沙土鼠全脑缺血３０分钟，再灌注１２０分钟的细胞外液中的谷胱甘肽（Ｇｌｕｔａｔｈｉｏｎｅ，ＧＳＨ）的变化及丹参对它的影响。结果显示：全脑缺血后，细胞外液ＧＳＨ水平迅速升高（Ｐ＜０．０１），缺血３０分钟达高峰为缺血前的５。８２倍。再灌注后ＧＳＨ水平明显降低，于３０分钟趋于正常。脑缺血前３０分钟给予丹参注射液不影响细胞外液ＧＳＨ水平。表明脑缺血及再灌注期，ＧＳＨ反应性增高。ＧＳＨ作为内源性抗氧化剂及ＮＭＤＡ受体拮抗剂在脑缺血损伤中起重要作用。