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Effects of Flower Buds Extract of Tussilago farfara on focal cerebral ischemia in Rats and Inflammatory Response in BV2 Microglia

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Chinese Journal of Integrative Medicine 2018年第11期24卷 844-852页

作者： Ji Hye Hwang Vinoth Kumar R Seok Yong Kang Hyo Won Jung Yong-Ki Park Department of Acupuncture and Moxibustion Medicine College of Korean Medicine Gachon University Department of Herbology College of Korean Medicine Dongguk University

Objective： To investigate the effects of the flower buds extract of Tussilago farfara Linné（Farfarae Flos; FF） on focal cerebral ischemia through regulation of inflammatory responses in activated microglia. Methods： Brain ischemia was induced in Sprague-Dawley rats by a transient middle cerebral artery occlusion（t MCAO） for 90 min and reperfusion for 24 h. Twenty rats were randomly divided into 4 groups（n=5 per group）： normal, t MCAO-induced ischemic control, t MCAO plus FF extract 300 mg/kg-treated, and t MCAO plus MK-801 1 mg/kg-treated as reference drug. FF extract（300 mg/kg, p.o.） or MK-801（1 mg/kg, i.p.） was administered after reperfusion. Brain infarction was measured by 2,3,5,-triphenyltetrazolium chloride staining. Neuronal damage was observed by haematoxylin eosin, Nissl staining and immunohistochemistry using anti-neuronal nuclei（Neu N）, anti-glial fibril ary acidic protein（GFAP）, and antiCD11 b/c（OX42） antibodies in ischemic brain. The expressions of inducible nitric oxide synthase（i NOS）, tumor necrosis factor（TNF-α）, and hypoxia-inducible factor-1 a（HIF-1α） were determined by Western blot. BV2 microglial cel s were treated with FF extract or its main bioactive compound, tussilagone with or without lipopolysaccharide（LPS）. Nitric oxide（NO） production was measured in culture medium by Griess assay. The expressions of i NOS, COX-2 and pro-inflammatory cytokines mRNA were analyzed by reverse transcription-polymerase chain reaction. The expression of i NOS, and COX-2 proteins, the phosphorylation of ERK1/2, JNK, and p38 MAPK and the nuclear expression of NF-κB p65 in BV2 cells were determined by Western blot. Results： FF extract significantly decreased brain infarctions in ischemic rats（P〈0.01）. The neuronal death and the microglia/astrocytes activation in ischemic brains were inhibited by FF extract. FF extract also suppressed i NOS, TNF-α, and HIF-1α expression in ischemic brains. FF extract（0.2 and 0.5 mg/m L, P〈0.01） and tussilagone 20 and 50 μmol/L, P〈0

关键词： Tussilago farfara focal cerebral ischemia inflammation microglia tussilagone

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EFFECT OF DL-3-N-BUTYLPHTHALIDE ON BRAIN EDEMA IN RATS SUBJECTED TO focal cerebral ischemia

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Chinese Medical Sciences Journal 1997年第2期12卷 102-106页

作者：邓文斌冯亦璞 Institute of Materia Medica CAMS & PUMC Beijing 100050

The present study evaluated the effect of dl-3-n-butylphthalide(NBP) ,a novel brain protective agent, on brain edema in rats following focal ischemia. Edema was induced by occluding the right middle cerebral artery (MCAO).producing permanent focal ischemia in the right cerebral hemisphere,which developed ip-silateral brain edema reproducibly. Edema was assessed 24 h after MCA occlusion by determining the brain water content from wet and dry weight measurements,and the sodium,potassium concentrations with ion-selective electrodes. In this model,NBP at the dose of 80,160 and 240 mg/kg po 15 min after MCAO prevented from brain edema in a dose-dependent manner. A significant reduction of sodium content and an increase in potassium level were observed in all drug-treated groups. It showed that NBP strongly attenuated brain water entry,sodium accumulation and potassium loss. Nimodipine treatment(5mg/kg sc) also reduced brain edema (P<0. 05). The results suggest that a strong anti-edema activity of NBP may play an important role to contribute to the treatment of ischemic damage.

关键词： dl-3-n-butylphthalide focal cerebral ischemia brain edema

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PROLIFERATION AND MIGRATION OF EPENDYMAL CELLS IN THE BRAIN OF ADULT RATS AFTER PERMANENT focal cerebral ischemia

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Journal of Pharmaceutical Analysis 2004年第1期16卷 62-66页

作者：张蓬勃刘勇李捷邱芬陈新林康前雁宋土生钱亦华薛荣亮 RongliangResearch Center 0f Neuroscience

Objective Ependymal cells are thought to be the primary source of neural stem cells in the adult central nervous system. The purpose of this study is to examine spatial and temporal profiles of ependymal cell proliferation and migration after focal cerebral ischemia. Methods Eighty male Sprague Dawley rats underwent permanent middle cerebral artery occlusion after injection of 10 μL of 0.2% Dil into the lateral ventricle. Rats were sacrificed and brain sections were acquired for pathological evaluation and laser confocal imaging at day 1,3,7,11,14,21 and 28 after ischemia. Results The density of Dil-labeled cells in the ischemic ipsilateral subventricular zone was significantly higher than that in the control group and these labeled cells dispersed in the ischemic ipsilateral subventricular zone and/or were located in ependyma from day 1 to 11. In the ischemic ipsilateral cortex, some Dil-labeled cells occurred in peri-infarction and infarction of parietal region at day14 and peaked at day 21 when some Dil-labeled cell nodules were found in this region. During postischemic day 14-28, a significant decrease in labeled cell density in the ischemic ipsilateral subventricular zone was coincident with a significant increase in labeled cells density in the cortex (peri-infarction and infarction). Conclusion The results indicate that ependymal cells proliferate and migrate after focal cerebral ischemia in the adult rat brain.

关键词： neurogenesis neural stem cells ependymal cells focal cerebral ischemia subventricular zone

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Expression profiles of microRNAs after focal cerebral ischemia/reperfusion injury in rats

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Neural Regeneration Research 2012年第12期7卷 917-923页

作者： Fengguo Zhai Xiuping Zhang Yue Guan Xudong Yang Yang Li Gaochen Song Lixin Guan Department of Pharmacology Mudanjiang Medical University Mudanjiang 157011 Heilongjiang Province China Department of Endocrinology Mudanjiang Medical University Mudanjiang 157011 Heilongjiang Province China Department of Thoracic Surgery Mudanjiang Medical University Mudanjiang 157011 Heilongjiang Province China Department of Biochemistry Mudanjiang Medical University Mudanjiang 157011 Heilongjiang Province China

Rat models of focal cerebral ischemia/reperfusion injury were established by occlusion of the middle cerebral artery. Microarray analysis showed that 24 hours after cerebral ischemia, there were nine up-regulated and 27 down-regulated microRNA genes in cortical tissue. Bioinformatic analysis showed that bcl-2 was the target gene of microRNA-384-5p and microRNA-494, and caspase-3 was the target gene of microRNA-129, microRNA-320 and microRNA-326. Real-time PCR and western blot analyses showed that 24 hours after cerebral ischemia, bcl-2 mRNA and protein levels in brain tissue were significantly decreased, while caspase-3 mRNA and protein levels were significantly increased. This suggests that following cerebral ischemia, differentially expressed microRNA-384-5p, microRNA-494, microRNA-320, microRNA-129 and microRNA-326 can regulate bcl-2 and caspase-3 expression in brain tissue.

关键词： focal cerebral ischemia microRNA apoptosis bcl-2 caspase-3 neural regeneration

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Caffeic acid ameliorates early and delayed brain injuries after focal cerebral ischemia in rats

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Acta Pharmacologica Sinica 2006年第9期27卷 1103-1110页

作者： Yu ZHOU San-hua FANG Yi-lu YE Li-sheng CHU Wei-ping ZHANG Meng-ling WANG Er-qing WEI~2 Department of Pharmacology,School of Medicine,Zhejiang University,Hangzhou 310031,China DepartmentofPharmacologySchoolofMedicine ZhejiangUniversityHangzhou310031China

Aim:To investigate the effects of caffeic acid on early and delayed injuries afterfocal cerebral ischemia in rats,and the possible relation to ***:Transient focal cerebral ischernia was induced by middlecerebral artery occlusion in Sprague-Dawley *** acid(10 and 50 mg/kg)was ip injected for 5 d after *** brain injuries were observed,and thelevels of cysteinyl leukotrienes and leukotriene B4in the brain tissue ***:Caffeic acid(50 mg/kg)ameliorated neurological dysfunctionand neuron loss,and decreased infarct volume 24 h after ischemia;it attenuatedbrain atrophy,infarct volume,and particularly astrocyte proliferation 14 d *** addition,it reduced the production of leukotrienes(5-lipoxygenasemetabolites)in the ischemic hemispheres 3 h and 7 d after ***:Caffeic acid has protective effect on both early and delayed injuries after focalcerebral ischemia in rats;and this effect may partly relate to 5-lipoxygenaseinhibition.

关键词： caffeic acid focal cerebral ischemia neuron astrocyte 5-lipoxygenase leukotriene

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Therapeutic window of Qingkailing injection for focal cerebral ischemia/reperfusion injury

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Neural Regeneration Research 2011年第21期6卷 1605-1611页

作者： Fafeng Cheng Wenting Song Xianggen Zhong Yi Lu Shaoying Guo Dong Wang Weipeng Zhao Qingguo Wang College of Basic Medicine Beijing University of Chinese Medicine Beijing 100029 China Xiyuan Hospital of China Academy of Chinese Medical Sciences Beijing 100091 China

The time window in which a drug is effective varies between drugs. The present study investigated the therapeutic window of Qingkailing injection for focal cerebral ischemia/reperfusion in mice. Animals underwent middle cerebral artery occlusion and were injected with Qingkailing （1.5, 3, 6 mL/kg）. Infarct volume and neurological function were assessed after 24 hours of ischemia. In addition, to establish the therapeutic time window, mice were injected with 3 mL/kg Qingkailing at 0, 1, 3, 4, 6, 9 and 12 hours after occlusion. Results revealed that Qingkailing injection significantly reduced infarct volume and improved neurological function in model mice after cerebral infarction for up to 9 hours, demonstrating that the therapeutic window of Qingkailing injection can extend to 9 hours for cerebral ischemia/reperfusion in mice.

关键词： focal cerebral ischemia Qingkailing injection time window infarct volume

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Changes in secretory pathway Ca^(2+)-ATPase 2 following focal cerebral ischemia/reperfusion injury

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Neural Regeneration Research 2013年第1期8卷 76-82页

作者： Tonglin Lu Zhiping Hu Liuwang Zeng Zheng Jiang Department of Neurology Second Xiangya Hospital Central South University

This study aimed to investigate changes in secretory pathway Ca2＋-ATPase 2 expression following cerebral ischemia/reperfusion injury, and to define the role of Ca2＋-ATPases in oxidative stress. A rat model of cerebral ischemia/reperfusion injury was established using the unilateral middle cerebral artery occlusion method. Immunohistochemistry and reverse transcription-PCR assay results showed that compared with the control group, the expression of secretory pathway Ca2＋-ATPase 2 protein and mRNA in the cerebral cortex and hippocampus of male rats did not significantly change during the ischemic period. However, secretory pathway Ca2＋-ATPase 2 protein and mRNA expression reduced gradually at 1, 3, and 24 hours during the reperfusion period. Our experimental findings indicate that levels of secretory pathway Ca2＋-ATPase 2 protein and mRNA expression in brain tissue change in response to cerebral ischemia/reperfusion injury.

关键词： neural regeneration brain injury cerebral infarction secretory pathway Ca2+-ATPase 2 Golgiapparatus Ca2+ oscillations manganese focal cerebral ischemia oxidative damage Ca2^-ATPase grants-supported paper photographs-containing paper neuroregeneration

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Aggravated inflammation and increased expression of cysteinyl leuko-triene receptors in the brain after focal cerebral ischemia in AQP4-deficient mice

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Neuroscience Bulletin 2012年第6期28卷 680-692页

作者： Wen-Zhen Shi Chun-Zhen Zhao Bing Zhao Qiao-Juan Shi Li-Hui Zhang Yan-Fang Wang San-Hua Fang Yun-Bi Lu Wei-Ping Zhang Er-Qing Wei Institute of Neuroscience and Department of Pharmacology School of MedicineZhejiang University Hangzhou Key Laboratory of Neurobiology and Department of Pharmacology School of Basic MedicineHangzhou Normal University

Objective Aquaporin-4 （AQP4）, the main water channel protein in the brain, plays a critical role in water homeostasis and brain edema. Here, we investigated its role in the inflammatory responses after focal cerebral ischemia. Methods In AQP4-knockout （KO） and wild-type mice, focal cerebral ischemia was induced by 30 rain of middle cerebral arterial occlusion （MCAO）. Ischemic neuronal injury and cellular inflammatory responses, as well as the expression and localization of cysteinyl leukotriene CysLT2 and CysLT~ receptors, were determined at 24 and 72 h after MCAO. Results AQP4-KO mice showed more neuronal loss, more severe microglial activation and neutrophil infiltration, but less astrocyte proliferation in the brain after MCAO than wild-type mice. In addition, the protein levels of both CysLT1 and CysLT2 receptors were up-regulated in the ischemic brain, and the up-regulation was more pronounced in AQP4-KO mice. The CysLT1 and CysLT2 receptors were primarily localized in neurons, microglia and neutrophils; those localized in microglia and neutrophils were enhanced in AQP4-KO mice. Conclusion AQP4 may play an inhibitory role in postischemic inflammation.

关键词： aquaporin 4 gene deficiency inflammation cysteinyl leukotriene receptor microglia astrocyte focal cerebral ischemia

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Effects of Fujian tablet on Nogo-A mRNA expression and plasticity of the corticospinal tract in a rat model of focal cerebral ischemia

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Neural Regeneration Research 2011年第33期6卷 2577-2581页

作者： Wei Liu Yonghong Zhou Qing Jia Bingbing Han Guoli Zhang Department of Encephalopathy Affiliated Hospital of Shandong University of Traditional Chinese Medicine Jinan 250011 Shandong Province China Department of Traditional Chinese Medicine Medical College of Qingdao University Qingdao 266071 Shandong Province China Institute of Basic Medicine Shandong Academy of Medical Science Jinan 250062 Shandong Province China College of Basic Medical Sciences Jinan 250355 Shandong Province China Department of Radiation Oncology Shandong Tumor Hospital ＆ Institute Jinan 250117 Shandong Province China

The present study investigated the effects of Fujian tablet, a Chinese medicine compound that can nourish liver and kidney, on corticospinal tract plasticity and cervical cord microenvironment in rats with focal cerebral ischemia. Results showed that motor function of rats with right proximal middle cerebral artery occlusion was significantly improved following treatment with Fujian tablet, 9 g crude drug/kg. Anterograde tracing revealed significantly increased biotinylated dextran amine expression in the denervated （left） side of the cervical cord （C4-6） following Fujian tablet treatment, and significantly decreased Nogo-A mRNA expression was detected in the denervated side of the cervical cord （C4-6） using in situ hybridization. Pearson＇s correlation analysis showed a negative correlation between biotinylated dextran amine and Nogo-A mRNA expression （r = -0.943, P 〈 0.01）. Results demonstrated that Fujian tablet can promote corticospinal tract plasticity possibly through the inhibitory effect on Nogo-A mRNA expression in the cervical spinal cord, thereby improving motor dysfunction.

关键词： Fujian tablet focal cerebral ischemia corticospinal tract Nogo-A mRNA Chinese herbal formula neural plasticity neural regeneration

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Hyperlipidemia affects neuronal nitric oxide synthase expression in brains of focal cerebral ischemia rat model

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Neural Regeneration Research 2008年第6期3卷 642-646页

作者： Jianji Pei Liqiang Liu Jinping Pang Xiaohong Tian Department of Internal Medicine Beijing Mentougou Hospital Beijing 102300 China Department of Neurology Third Affiliated Hospital Inner Mongolia Medical College Baotou 014010 Nei Monggol Autonomous Region China Department of Neurology First Affiliated Hospital Shanxi Medical University Taiyuan 030001 Shanxi Province China

BACKGROUND： Hyperlipidemia, a risk factor for ischemic cerebrovascular disease, may mediate production of neuronal nitric oxide synthase （nNOS） to induce increased nitric oxide levels, resulting in brain neuronal injury. OBJECTIVE： To investigate effects of hyperlipidemia on brain nNOS expression, and to verify changes in infarct volume and pathology during reperfusion, as well as neuronal injury following ischemia/reperfusion in a rat model of focal cerebral ischemia. DESIGN, TIME AND SETTING： Complete, randomized grouping experiment was performed at the Laboratory of Physiology, Shanxi Medical University from March 2005 to March 2006. MATERIALS： A total of 144 eight-week-old, male, Wistar rats, weighing 160-180 g, were selected. A rat model of middle cerebral artery occlusion was established by suture method after 4 weeks of formulated diet. Nitric oxide kit and rabbit anti-rat nNOS kit were respectively purchased from Nanjing Jiancheng Bioengineering Institute, China and Wuhan Boster Biological Technology, Ltd., China. METHODS： The rats were equally and randomly divided into high-fat diet and a normal diet groups. Rats in the high-fat diet group were fed a high-fat diet, consisting of 10% egg yolk powder, 5% pork fat, and 0.5% pig bile salt combined with standard chow to create hyperlipidemia. Rats in the normal diet group were fed a standard rat chow. A total of 72 rats in both groups were randomly divided into 6 subgroups： sham-operated, 4-hour ischemia, 4-hour ischemia/2-hour reperfusion, 4-hour ischemia/4-hour reperfusion, 4-hour ischemia/6-hour reperfusion, and 4-hour ischemia/12-hour reperfusion, with 12 rats in each subgroup. MAIN OUTCOME MEASURES： nNOS expression was measured by immunohistochemistry, and pathomorphology changes were detected by hematoxylin-eosin staining. Infarct volume and nitric oxide levels were respectively measured using 2, 3, 5-triphenyltetrazolium chloride （TTC） and immunohistochemistry. RESULTS： In the ischemic region, pathology c

关键词： focal cerebral ischemia hyperlipidemia ischemia/reperfusion injury neuronal nitric oxides ynthase

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