Mechanism of JAK2-STAT3 signaling pathway in ischemic postconditio-ning neuroprotection after cerebral ischemia in tree shrews

AIM: To investigate the regulatory effect of JAK2-STAT3 signaling pathway on the neuroprotection of ischemic postconditioning (IPoC) in tree shrews, and to explore the mechanisms of cerebral injury deterioration after inhibiting the JAK2-STAT3 pathway. METHODS: The model of thrombotic cerebral ischemia was induced by photochemical reaction in tree shrews and the IPoC was established at 4 h after ischemia followed by clipping ipsilateral common carotid artery on the ischemia side for 5 min (3 times). After IPoC and intracerebroventricular injection of AG490 (JAK2 inhibitor), the changes of cerebral infarction area were detected by TTC staining, and the histological and ultrastructural changes of cortical neurons were observed under light and electron microscopes, respectively. The protein levels of t-STAT3 and p-STAT3 in the cortical tissue were determined by Western blot. RESULTS: The neuronal pycnosis, mitochondrial swelling and vanish of the mitochondrial cristae were found in cortical cortex, and the infarction area was (24.78±3.30)% at 24 h after cerebral ischemia. Meanwhile, the phosphorylation level of STAT3 protein in the cortical tissue was significantly increased (P<0.01). The cortical neuronal damage and mitochondrial swelling were decreased after IPoC, the area of cerebral infarction was significantly reduced to (17.67±1.83)% (P<0.01), and the phosphorylation level of STAT3 protein was further increased (P<0.01). However, the neuronal damage was aggravated, the infarction area was expanded to (23.85±2.77)%(P<0.05) after treatment with AG490, and the phosphorylation level of STAT3 protein was also significantly reduced (P<0.05). CONCLUSION: IPoC may reduce cerebral injury by regulating the phosphorylation of STAT3 protein, and inhibition of JAK2-STAT3 signaling pathway may counteract the cerebral protective effect of IPoC and aggravate brain injury.     

Effects of ischemic postconditioning on hippocampus CA1 area rCBF and astrocyte activation after cerebral ischemia in tree shrews

AIM: To study the effects of ischemic postconditioning (PC) on regional cerebral blood flow (rCBF) and astrocyte (AS) activation in hippocampus CA1 area and to explore the possible mechanism of ischemic PC affecting glial fibrillary acidic protein (GFAP) expression during focal cerebral thrombosis. METHODS: The thrombotic focal cerebral ischemia was induced by photochemical reaction in tree shrews, and ischemic postconditioning was established by cliped ipsilateral carotid of the animal at 4 h after cerebral ischemia. The rCBF and GFAP expressions in hippocampus CA1 area were detected, respectively, by laser-Doppler (LD) fowmeter and immunohistochemistry. RESULTS: The numbers of GFAP positive cells were increased markedly and GFAP expression enhanced (P<0.01). AS oncosis was apparent 24 h after cerebral ischemia. Postconditioning increased hippocampus rCBF from (2.55±0.28) PU to (10.42±3.75) PU (P<0.05) at 24 h and from (9.84±1.22) PU to (18.74±1.60) PU (P<0.05) at 72 h after the cerebral ischemia, and AS oncosis was inhibited markedly. CONCLUSION: Multiple, short, regional carotid occlusions may prolong “time window” of therapeutic cerebral ischemia. The protection mechanism of the ischemic postconditioning may be associated with the increase in rCBF and improvement of hippocampus microenvironment by regulating AS activation.     

稻田上覆水中光致活性组分形成过程及其环境效应研究进展

稻田上覆水中富含有机质、亚硝酸盐和硝酸盐等光敏活性物质,在太阳光作用下会产生三重激发态有机质(3CDOM*)、单线态氧(1O2)和羟基自由基(?OH)等活性组分,其对稻田污染物转化和碳氮等元素循环具有重要意义。基于此,综述了稻田上覆水中光致活性组分产生过程和机制,重点介绍了水稻不同生长周期内光活性组分的类型、通量变化趋势;探讨了不同环境因素对自由基产生的影响;阐述了上覆水光致活性组分对稻田中砷和不同有机污染物的非生物转化贡献与机制,并展望了稻田上覆水光化学过程的未来研究方向。     

Effects of hyperglycemia on ionic homeostasis in hippocampal microenvironment and secondary neuronal injury after focal ischemia in cerebral cortex of tree shrews

AIM：To observe the effects of hyperglycemia on the ionic homeostasis in hippocampal microenvironment after thrombotic cortical ischemia in tree shrews, and to explore the action and mechanisms of hyperglycemia in secondary neuronal injury after ischemia. METHODS：High blood glucose in tree shrews was induced by intraperitoneal injection of streptozocin. Focal thrombotic cortical ischemia was induced by photochemical method in tree shrews. At 4, 24 and 72 h after ischemia, the changes of pH, K+, Na+, Ca2+ and Cl- in the ipsilateral ischemic hippocampal microenvironment were tested by a single-pumped push-pull microdialysis system and an ion analyzer. The histopathological changes and hippocampal neuronal density were also examined. RESULTS：After cortical ischemia in tree shrews, the pH and the concentrations of Na+, Ca2+ and Cl- in the hippocampal microenvironment decreased, while the concentration of K+ increased. These differences were the most significant at 4 h, the second at 24 h and insignificant at 72 h. Combination of hyperglycemia and cerebral ischemia worsened the turbulence of ionic homeostasis. Compared with the normoglycemic ischemic animals, the changes of pH, K+ and Ca2+ concentrations at 4 h as well as pH and Na+ at 24 h in the hyperglycemic ischemic animals were more significant (P<0.05). The results of histopathological examination showed that there was ischemic neuronal damage in the exposed cerebral cortex and the ipsilateral hippocampal CA1 region at 4 h after photochemical reaction, and the damage was the most severe at 24 h, subsequently accompanied with glial proliferation at 72 h. The hyperglycemic ischemic animals suffered from greater neuronal injury in the cortex and hippocampus than the normoglycemic ischemic animals, especially at 24 h (P<0.01) and 72 h (P<0.05). CONCLUSION: The disturbance of acid-base equilibrium and ionic homeostasis in hippocampal microenvironment, following the spreading of the microenvironment in ischemic core, may be an important reason for secondary neuronal injury in the hippocampus after thrombotic cortical ischemia in tree shrews. Hyperglycemia aggravates the turbulence of ischemic ionic microenvironment.     

Changes in platelet-activating factor receptor binding characteristics in cerebral thrombotic core and penumbra of tree shrews

AIM: To observe the changes in platelet-activating factor (PAF) receptor binding characteristics and explore the action of PAF on formation of thrombotic core and penumbra following local cerebral ischemia. METHODS: Neuron's membrane protein was abstracted, and the local cerebral ischemia model were induced by photochemistry in tree shrews. The PAF binding sites on central neuron membrane were studied by-PAF binding assay. RESULTS: There were two different affinities of PAF receptors on tree shrew's brain cell membrane, with kD₁=(3.61 ±0.72) nmol/L and kD₂=(17.04±2.41) nmol/L, corresponding respectively to maximum number of binding sites: B_max1=(1 457.94±168.01) pmol/g protein and B_max2=(5 017.40±742.16) pmol/g protein. The binding sites decreased in ischemic core, penumbra and contralateral regions at 4,24 and 72 h after ischemia (P<0.01), with those of 24 h reaching the minimum levels. CONCLUSION: PAF receptors play an important role in cerebral ischemia, may be related to the secondary damage in ischemic penumbra, and also are molecular bases of brain injury induced by PAF.     

玉米持绿与早衰品种叶片衰老过程中光化学活性的变化

为了探讨不同衰老型玉米叶片在衰老过程中光化学反应及其对光合能力维持的贡献, 本研究使用持绿玉米品种“齐319”和早衰玉米品种“黄早四”, 在控制的条件下, 用乙烯利诱导离体叶片衰老, 通过快速叶绿素荧光诱导动力学曲线(OJIP曲线)和820 nm光吸收等技术, 研究了衰老过程中叶片叶绿素含量、光合速率、光系统I (PSI)、光系统II (PSII)以及光合电子传递体活性的变化。结果表明, 在衰老过程中, 齐319叶片叶绿素含量和光合速率的下降速度明显慢于黄早四, 是功能型持绿品种。玉米叶片的衰老伴随着OJIP曲线的J、I、K、L点荧光的增加, 以及远红光诱导820 nm光信号落差的下降。与齐319相比, 黄早四叶片的OJIP曲线和820 nm光吸收曲线的变化更剧烈。我们认为, 在衰老过程中PSI和PSII光化学活性的快速下降和光合电子传递功能的衰退是玉米叶片光合能力迅速下降的重要原因之一。在此基础上, 讨论了衰老过程中玉米叶片中与光合作用有关蛋白与光合能力下降的可能关系。     

去果、枝条环剥和新梢套袋后桃树叶片可溶性糖含量的变化对光合作用的影响

以‘绿化九号’桃(Prunus persica(L.) Batch)为试材,在果实最后迅速生长期,通过去果降低叶片中糖分往果实中的运输,新梢套袋等措施改变光照时间,减少光合同化物的积累,并通过新梢左右两端环剥阻止同化物向其他果实和枝条运输,形成一个叶片中糖分浓度梯度,研究不同叶片糖分浓度梯度下,源叶净光合速率(Pn)、叶绿素荧光、叶片中可溶性糖含量等的变化。同时,通过叶绿素荧光参数分析了在库源调控造成叶片可溶性含量不同时,Pn下降可能的调控机制。结果表明,和留果对照相比,去果造成叶片可溶性糖含量最高,而套袋使叶片可溶性糖含量最低。伴随着叶片可溶性糖含量的增加,Pn和气孔导度(Gs)下降,而叶片温度(tleaf)增加。Pn和Gs之间极显著正相关。而tleaf的增加则伴随着光系统Ⅱ最大量子产额(Fv/Fm)、光系统Ⅱ实际光化学效率(ΦPSⅡ)和光化学猝灭系数(qP)的下降,相反,tleaf增加,非光化学猝灭系数(NPQ)增加。因而,叶片可溶性糖的积累有可能通过某种方式影响Gs并使之降低,造成蒸腾作用减少,最后使叶片tleaf升高,有可能限制了PSⅡ反应中心的光化学效率,从而对光合作用做出调节。     

Changes in monoamine oxidase activity during thrombotic cerebral ischemia and protection mechanisms of ginkgolide B in tree shrews

AIM:The present study was designed to examine changes in monoamine oxidase (MAO) activity during cerebral ischemia and whether ginkgolide B's brain protection challenges with inhibiting monoamine oxidase. METHODS: The focal thrombotic cerebral ischemia was formed by photochemistry-induced in tree shews.MAO activities in different areas which include ischemic,core, penumbra and contralater and serum, were tested by enzyme color-compared way. The protein contents in different area above was examined by amino acid autoanalytic apparatus. RESULTS:MAO activities in ischemic core in different group were much lower than that in the sham operation group and contralatetral areas, with its peak at seventy-two hours after occlusion, but that in penumbra and serum ascended. There were significant differences in MAO activities between ischemic group and control (P＜0.01). In ginkgolide B(GB) group, the MAO activities in all areas but not in core descended, significant differences(P＜0.01) between in GB group and in twenty-four hours after occlusion. Changes in MAO activity was consistent with alterations of brain proein content(r=0.81,P＜0.05). CONCLUSION:The changes in monoamine neurotransmitters in core and penumbra considerably depend on the alterations of MAO activities after thrombotically cerebral ischemia. Probably, protective effects of GB on ischemic neurons is related to its acting as antagonist of platelet activating factor and regulator of monoamine oxidase.     

Ischemia postconditioning induces tight junction protein expression and inhibits brain edema after thrombotic cerebral ischemia in tree shrews

AIM: To assess whether the expression of tight junction(TJ) proteins, occludin/zonula occludins(ZO)-1, and regional cerebral blood flow(rCBF) link to brain edema in tree shrews during thrombotic cerebral ischemia and ischemic postconditioning(PC), and to explore how TJ affects brain edema and cerebral infarction. METHODS: Tree shrews were randomly grouped into control, ischemia and cerebral ischemia+PC(n=23), and the remaining 3 animals were used for magnetic resonance imaging(MRI). The local cerebral thrombosis were induced by photochemical reaction in the tree shrews, and ischemic PC was established at 4 h after induction of cerebral ischemia followed by clipped ipsilateral common carotid artery(5 min×3). The changes of the neural ultrastructure were observed under electron microscope. The neuronal apoptosis was analyzed by the method of TUNEL. Laser Doppler brain flowmetry was used to monitor the rCBF. The protein levels of occludin/ZO-1 were determined by immunochemistry and Western blot. The cerebral infarction volume was detected by MRI. The brain water content was measured by dry-wet weight method. RESULTS: Induction of cerebral ischemia led to a significant reduction of the normal neuron numbers in the hippocampal CA1 area, and conversely, the number of neurons with abnormal ultrastructure was increased. The TUNEL positive cells were increased significantly(P<0.01) in ischemia group. Moreover, the rCBF decreased significantly(P<0.01), and occludin/ZO-1 protein expression decreased(P<0.01). The brain water content and cerebral infarction volume were significantly increased(P<0.01). Ischemic PC increased the rCBF and the occludin/ZO-1 expression, but reduced the brain water content, the TUNEL positive cells, and the infarction volume(P<0.01). CONCLUSION: Ischemic PC increases the rCBF but not the local water content, suggesting that reduced cerebral infarction volume after ischemia PC is associated with the attenuation of cerebral edema by the enhancement of occludin/ZO-1 protein expression.