Effect of Tongxinluo on basic fibroblast growth factor mRNA after cerebral ischemia/reperfusion injury

INTRODUCTION Neural stem cells (NSCs) are characterized by self-renewing and mul- ti-differentiated potencies. Exogenous factors, such as growth factors and cytokines, have great effects on the proliferation and differentia- tion of NSCs. Some researches …     

Expression of anti-apoptotic protein Bcl-2 and proapoptotic protein Bax after delayed paraplegia induced by ischemia/reperfusion injury

INTRODUCTION Apoptosis is known to occur in the central nervous system during de- velopment and in pathological settings such as ischemia reperfusion (IR) injury[1]. Apoptosis requires an active commitment of the cell to degrade its own DNA, according to …     

Cromakalin pretreatment affects mitochondrial structure and function in a rat model of ischemia/reperfusion injury☆

BACKGROUND：Mitochondrial structural changes and energy dysmetabolism frequently occur subsequent to cerebral ischemia.Adenosine triphosphate（ATP）-sensitive potassium channel openers exhibit protective effects on cerebral ischemia/reperfusion injury.OBJECTIVE：To validate the effects of cromakalin on mitochondrial structure and function in ischemic penumbra brain tissue in a rat model of middle cerebral artery occlusion（MCAO）.DESIGN,TIME AND SETTING：The present single-factor analysis of variance,randomized,controlled,animal experiment was performed at the Institute of Brain Science,Affiliated Hospital of Qingdao University Medical College between October 2007 and March 2008.MATERIALS：Forty male,Wistar rats were randomly divided into four groups,with 10 rats per group：sham-operated,MCAO,MCAO＋ATP-sensitive potassium channel opener（cromakalin）,and MCAO＋cromakalin＋ATP-sensitive potassium channel blocking agent（glibenclamide）.METHODS：Focal cerebral ischemia/reperfusion injury was induced by MCAO in all groups except the sham-operated group.The MCAO cromakalin group was administered 10 mg/kg cromakalin（i.p.） prior to MCAO induction.The MCAO＋cromakalin＋glibenclamide group received an injection of 10 mg/kg cromakalin（i.v.）,and subsequently an injection of 10 mg/kg cromakalin（i.p.） prior to MCAO induction.MAIN OUTCOME MEASURES：At 24 hours after cerebral ischemia/reperfusion injury,cellular apoptosis was detected by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate（dUTP） nick-end labeling technique.Cytochrome C expression was measured by immunohistochemistry.In addition,mitochondrial swelling,membrane fluidity,membrane phospholipid and malonaldehyde（MDA） contents,as well as Na^＋-K^＋-ATPase,Ca^2 ＋-ATPase,and superoxide dismutase（SOD） activities were determined.RESULTS：Compared with the sham-operated group,the three ischemia groups exhibited significantly elevated mitochondrial MDA content,reduced membrane phospholipid and ATP conten     

Matrix metalloproteinase-9 expression and blood brain barrier permeability in the rat brain after cerebral ischemia/reperfusion injury**☆

BACKGROUND： The integrity of the blood brain barrier （BBB） plays an important role in the patho-physiological process of cerebral ischemia/reperfusion injury. It has been recently observed that metalloproteinase-9 （MMP-9） is closely related to cerebral ischemia/reperfusion injury OBJECTIVE： This study was designed to observe MMP-9 expression in the rat brain after cerebral ischemia/reperfusion injury and to investigate its correlation to BBB permeability. DESIGN, TIME AND SETTING： This study, a randomized controlled animal experiment, was performed at the Institute of Neurobiology, Central South University between September 2005 and March 2006. MATERIALS： Ninety healthy male SD rats, aged 3-4 months, weighing 200-280 g, were used in the present study. Rabbit anti-rat MMP-9 polyclonal antibody （Boster, Wuhan, China） and Evans blue （Sigma, USA） were also used. METHODS： All rats were randomly divided into 9 groups with 10 rats in each group： normal control group, sham-operated group, and ischemia for 2 hours followed by reperfusion for 3, 6, 12 hours, 1, 2, 4 and 7 days groups. In the ischemia/reperfusion groups, rats were subjected to ischemia/reperfusion injury by suture occlusion of the right middle cerebral artery. In the sham-operated group, rats were merely subjected to vessel dissociation. In the normal control group, rats were not modeled. MAIN OUTCOME MEASURES： BBB permeability was assessed by determining the level of effusion of Evans blue. MMP-9 expression was detected by an immunohistochemical method. RESULTS： All 90 rats were included in the final analysis. BBB permeability alteration was closely correlated to ischemia/reperfusion time. BBB permeability began to increase at ischemia/reperfusion for 3 hours, then it gradually reached a peak level at ischemia/reperfusion for 1 day, and thereafter it gradually decreased. MMP-9 expression began to increase at ischemia/reperfusion for 3 hours, then gradually reached its peak level 2 days after perfusion, and thereafter it grad     

Effect of lidocaine on retinal aquaporin-4 expression after ischemia/reperfusion injury in the rat★

BACKGROUND: Several studies have demonstrated that high doses of lidocaine can reduce edema in rats with brain injury by down-regulating aquaporin-4 (AQP4) expression. The hypothesis for the present study is that lidocaine could retinal edema that is associated with AQP4 expression.OBJECTIVE: This study was designed to investigate the interventional effects of lidocaine on retinal AQP4 expression and retinal edema following ischemia/reperfusion injury in the rat.DESIGN, TIME AND SETTING: This study, a randomized, controlled, animal experiment, was performed at the Basic Research Institute, Chongqing Medical University from September 2006 to May 2007.MATERIALS: Seventy-five, healthy, adult, female, Sprague-Dawley rats were included. A total of 50 rats were used to establish a retinal ischemia/reperfusion injury model using an anterior chamber enhancing perfusion unit. Rabbit anti-rat AQP4 antibody was purchased from Santa Cruz Biotechnology, USA.METHODS: All 75 rats were randomly divided into three groups, with 25 rats in each: control, model, and lidocaine. At each time point (1, 6, 12, 24, and 48 hours after modeling, five rats for each time point), each rat in the lidocaine group was intraperitoneally administered lidocaine with an initial dose of 30 mg/kg, followed by subsequent doses of 15 mg/kg every six hours. The entire treatment process lasted three days for each rat. At each above-mentioned time point, rats in the model group were modeled, but not administered any substances. Rats in the control group received the same treatments as in the lidocaine group except that lidocaine was replaceld by physiological saline.MAIN OUTCOME MEASURES: Following hematoxylin-eosin staining, rat retinal tissue was observed to investigate retinal edema degree through the use of an optical microscope and transmission electron microscope. Retinal AQP4 expression was determined by immunohistochemistry.RESULTS: At each above-mentioned time point, AQP4 expression was significantly increased in the model group compared to the control group (P<0.05); this change was consistent with the degree of retinal edema. In the lidocaine group, retinal AQP4 expression was significantly decreased (P<0.05), and retinal edema was reduced, compared with the model group.CONCLUSION: Lidocaine inhibits rat retinal AQP4 expression following ischemia/reperfusion injury, leading to a reduction of retinal edema.     

Effect of lidocaine on retinal aquaporin-4 expression after ischemia/reperfusion injury in the rat★

BACKGROUND: Several studies have demonstrated that high doses of lidocaine can reduce edema in rats with brain injury by down-regulating aquaporin-4 (AQP4) expression. The hypothesis for the present study is that lidocaine could retinal edema that is associated with AQP4 expression. OBJECTIVE: This study was designed to investigate the interventional effects of lidocaine on retinal AQP4 expression and retinal edema following ischemia/reperfusion injury in the rat. DESIGN, TIME AND SETTING: This study, a randomized, controlled, animal experiment, was performed at the Basic Research Institute, Chongqing Medical University from September 2006 to May 2007. MATERIALS: Seventy-five, healthy, adult, female, Sprague-Dawley rats were included. A total of 50 rats were used to establish a retinal ischemia/reperfusion injury model using an anterior chamber enhancing perfusion unit. Rabbit anti-rat AQP4 antibody was purchased from Santa Cruz Biotechnology, USA. METHODS: All 75 rats were randomly divided into three groups, with 25 rats in each: control, model, and lidocaine. At each time point (1, 6, 12, 24, and 48 hours after modeling, five rats for each time point), each rat in the lidocaine group was intraperitoneally administered lidocaine with an initial dose of 30 mg/kg, followed by subsequent doses of 15 mg/kg every six hours. The entire treatment process lasted three days for each rat. At each above-mentioned time point, rats in the model group were modeled, but not administered any substances. Rats in the control group received the same treatments as in the lidocaine group except that lidocaine was replaceld by physiological saline. MAIN OUTCOME MEASURES: Following hematoxylin-eosin staining, rat retinal tissue was observed to investigate retinal edema degree through the use of an optical microscope and transmission electron microscope. Retinal AQP4 expression was determined by immunohistochemistry. RESULTS: At each above-mentioned time point, AQP4 expression was significantly increased in the model group compared to the control group (P < 0.05); this change was consistent with the degree of retinal edema. In the lidocaine group, retinal AQP4 expression was significantly decreased (P < 0.05), and retinal edema was reduced, compared with the model group. CONCLUSION: Lidocaine inhibits rat retinal AQP4 expression following ischemia/reperfusion injury, leading to a reduction of retinal edema. Key Words: ischemia/reperfusion; lidocaine; aquaporin-4     

Dose-dependent effects of procyanidin on nerve growth factor expression following cerebral ischemia/ reperfusion injury in rats★

BACKGROUND： Recently, grape seed procyanidin （GSP） has been shown to be exhibit antioxidant effects, effectively reducing ischemia/reperfusion injury and inhibiting brain cell apoptosis. OBJECTIVE： To study the effects of GSP on nerve growth factor （NGF） expression and neurological function following cerebral ischemia/reperfusion injury in rats. DESIGN： Randomized controlled study based on SD rats. SETTING： Weifang Municipal People＇s Hospital. MATERIALS： Forty-eight healthy adult SD rats weighing 280-330 g and irrespective of gender were provided by the Experimental Animal Center of Shandong University. GSP derived from grape seed was a new high-effective antioxidant provided by Tianjin Jianfeng Natural Product Researching Company （batch number： 20060107）. Rabbit-anti-rat NGF monoclonal antibody was provided by Beijing Zhongshan Biotechnology Co., Ltd., and SABC immunohistochemical staining kit by Wuhan Boster Bioengineering Co., Ltd. METHODS： The present study was performed in the Functional Laboratory of Weifang Medical College from April 2006 to January 2007. Forty-eight SD rats were randomly divided into the sham operation group, ischemia/reperfusion group, high-dose GSP （40 mg/kg） group, or low-dose GSP （10 mg/kg） group （n = 12 per group）. Ischemia/reperfusion injury was established using the threading embolism method of the middle cerebral artery. Rats in the ischemia/reperfusion model group were given saline injection （2 mL/kg i.p.） once daily for seven days pre-ischemia/reperfusion, and once more at 15 minutes before reperfusion. Rats in the high-dose and low-dose GSP groups were injected with GSP （20 or 5 mg/mL i.p., respectively, 2 mL/kg） with the same regime as the ischemia/reperfusion model group. The surgical procedures in the sham operation group were as the same as those in the ischemia/reperfusion model group, but the thread was approximately 10 mm long, thus, the middle cerebral artery was not blocked. MAIN OUTCOME MEASURES： NGF expression in the     

Effects of Rubus parvifolius L. on neuronal apoptosis and expression of apoptosis-related proteins in a rat model of focal cerebral ischemic-reperfusion injury*★

BACKGROUND： Studies have shown that the Rubus parvifolius L. （RP） plant extract exhibits protective effects on cerebral ischemia. This effect is reflected in altered ischemic neuronal apoptosis and associated protein expression.
OBJECTIVE： To explore the neuroprotective mechanism of RP after cerebral ischemia injury.
DESIGN, TIME AND SETTING： Randomized control experiment of cellular, molecular, and protein levels. The experiment was completed at Chongqing Medical University at the School of Pharmacy Laboratories and Basic Medical Institute from October 2005 to January 2006.
MATERIALS： Twenty-four adult, male, Wistar rats, weighing （28 ± 20） g. RP extract, which was a product of ethanol extraction, was provided by the Laboratory of Pharmaceutical Analysis, Chongqing Medical University. RP was dissolved in distilled water to a concentration of 10 mg/mL. All rats were randomly assigned into four groups： 5 g/kg RP, 10 g/kg RP, model, and sham-surgery, with 6 rats in each group.
METHODS： In the 5 and 10 g/kg RP groups, as well as the model group, the middle cerebral artery was occluded （MCAO） for 60 minutes, resulting in focal cerebral ischemia, followed by reperfusion for 24 hours. Rats in the 5 and 10 g/kg RP groups received 5 and 10 g/kg RP, respectively. The RP treatment group received RP intragastrically （once a day） for 3 days. One hour after the last dose, rats were subjected to MCAO. The same surgical procedure was performed in the sham-surgery group, except the suture was introduced into the external carotid artery, but not advanced. Rats in the model group were subjected to MCAO. The sham-surgery and model groups received intragastrically administered normal saline once per day for 3 days. One hour after the last dose, the rats were subjected to surgery.
MAIN OUTCOME MEASURES： TUNEL labeling and immunohistochemical methods were used to investigate changes in neuronal apoptosis and expression of the apoptosis-related proteins, Bax and Bcl-2, on the ischemic hemisphere     

Changes in corticocerebral morphology in a rat model of focal cerebral ischemia/reperfusion injury following ＂Xingnao Kaiqiao＂ acupuncture

BACKGROUND： Cerebral ischemia/reperfusion injury has been shown to induce inflammatory reactions, including white blood cell activation and adhesion molecule expression. These reactions often lead to aggravated neuronal injury. OBJECTIVE： To observe corticocerebral pathology, as well as ultrastructural changes, in a rat model of focal cerebral ischemia/reperfusion injury through optical and electron microscopy, and to investigate interventional effects of ＂Xingnao Kaiqiao＂ acupuncture （a brain-activating and orifice-opening acupuncture method）. DESIGN, TIME AND SETTING： A randomized, controlled, neuropathology, animal experiment was performed at the Laboratory of Molecular Biology, First Affiliated Hospital of Tianjin University of Traditional Chinese Medicine between April and June 2004. MATERIALS： A total of 50 healthy, male, Wistar rats were randomized into 5 groups, with 10 rats per group： control, sham-operated, model, non-acupoint, and ＂Xingnao Kaiqiao ＂. Transmission electron microscope （TEM 400ST） was provided by Philips, Netherlands. Electro-acupuncture treatment apparatus （KWD-8082） was provided by Changzhou Wujin Great Wall Medical Instrument, China. METHODS： Focal cerebral ischemia/reperfusion injury was induced by occlusion of the middle cerebral artery in the model, non-acupoint, and ＂Xingnao Kaiqiao＂ groups. Rats from the control group did not undergo any treatment. The sham-operated group received identical experimental procedures as the model group, except that the nylon suture was not inserted into the right internal carotid artery. At 1, 3, 6, and 12 hours following focal cerebral ischemia/reperfusion injury induction, rats from the Xingnao Kaiqiao group underwent 1-minute acupuncture at the bilateral ＂Neiguan＂ （PC 6） acupoint, using a reducing method of lifting-thrusting and twirling-rotating. Subsequently, the rats were subjected to acupuncture at the ＂Renzhong＂ （DU26） acupoint 10 times by a heavy bird-pecking method. The non-acupoint group     

Effects of exogenous ganglioside-1 on learning and memory in a neonatal rat model of hypoxia-ischemia brain injury★

BACKGROUND: Exogenous ganglioside-1 (GM1) can cross the blood-brain barrier and play a protective role against hypoxia-ischemia-induced brain damage. OBJECTIVE: To examine the possible mechanisms of exogenous GM1 protection in hypoxia-ischemia-induced brain damage in a neonatal rat model by measuring changes in brain mass, pathological morphology, growth-associated protein-43 expression, and neurobehavioral manifestations. DESIGN, TIME AND SETTING: A randomized block-design study was performed at the Immunohistochemistry Laboratory of the Pediatric Research Institute, Children’s Hospital of Chongqing Medical University from August 2005 to August 2006. MATERIALS: A total of 36 neonatal, 7-day-old, Sprague Dawley rats were used in this experiment. The hypoxia-ischemia-induced brain damage model was established by permanently occluding the right carotid artery, followed by oxygen inhalation at a low concentration (8% O2, 92% N2) for 2 hours. METHODS: All rats were randomly divided into the following groups: GM1, model, and sham operation, with 12 rats each group. Rats in the GM1 and model groups received hypoxic/ischemic-induced brain damage. Rats in the GM1 group received injections of GM1 (i.p., 20 mg/kg) at 0, 24, 48, 72, 96, 120, and 144 hours following models established, and rats in the model group were administered (i.p.) the same amount of saline. The right carotid artery was separated, but not ligated, in the sham operation group rats. MAIN OUTCOME MEASURES: At 1 week after surgery, expression of growth-associated protein-43, a marker of neural development and plasticity, was detected in the hippocampal CA3 region by immunohistochemistry. Brain mass was measured, and the pathological morphology was observed. At 4 weeks after surgery, behavioral changes in the remaining rats were tested by Morris water maze, and growth-associated protein-43 expression was measured. RESULTS: (1) In the GM1 and sham operation groups, growth-associated protein-43 expression was greater in the hippocampal CA3 region compared to the model group 1 week after surgery (P < 0.05). In all three groups, brain weight of the right hemisphere was significantly less than the left hemisphere, in particular in the model group (P < 0.05). In the GM1 group, the weight difference between two hemispheres, as well as the extent of damage in the right hemisphere, was less than the model group (P < 0.01). In the sham operation group, brain tissue consisted of integrated structures and ordered cells. In the model group, the cerebral cortex layers of the right hemisphere were not defined, neurons were damaged, and neurons were disarranged in the hippocampal area. In the GM1 group, neurons were dense in the right cerebral cortex and hippocampal area, with no significant change in glial proliferation. (2) The average time of escape latency in the GM1 group was shortened 4 weeks after surgery, and significantly less than the model group (P < 0.05). In addition, the frequency platform passing in the GM1 group was significantly greater than the model group (P < 0.01). CONCLUSION: Exogenous GM1 may reduce brain injury and improve learning and memory in hypoxia-ischemia-induced brain damage rats. This protection may be associated with increased growth-associated protein-43 expression, which is involved in neuronal remodeling processes. Key Words: ganglioside; growth-associated protein-43; hypoxia-ischemia brain damage; Morris water maze     

Effect of compound preparation Tongqiao Jiannao capsules on neural cell apoptosis and Bcl-2 and Bax protein levels in a rat model of brain ischemia/reperfusion injury

BACKGROUND:Pharmacological studies have demonstrated that compound preparation Tongqiao Jiannao capsules composed of Zexie, Baizhu, Honghua, Danshen, and Shexiang can supplement qi, activate blood circulation, relieve blood stasis, induce resuscitation for alleviating pain, relieve pain, and dilate blood vessels. OBJECTIVE: To observe the effects of Tongqiao Jiannao capsules on the levels of the anti-apoptotic protein Bcl-2 and the proapoptotic protein Bax, and verify the mechanism of action. DESIGN, TIME A...     

大鼠全脑缺血再灌注损伤后调控基因Bcl-2、Bax的表达及与细胞凋亡的关系

目的:本研究旨在探讨Bcl-2及Bax蛋白在大鼠全脑缺血再灌注损伤中的变化及与细胞凋亡的关系。方法:雄性Wistar大鼠56只,随机分为假手术组、缺血15分钟再灌注1、6、12、24、48、72小时组。采用大鼠四条血管阻断方法制备大鼠全脑缺血再灌注模型。采用TUNEL法观察不同再灌注时间组海马CAl区细胞凋亡的变化。采用免疫组化法观察Bcl-2及Bax蛋白表达水平的变化。结果:脑缺血损伤后随再灌注时间延长凋亡细胞逐渐增多,至再灌注48小时达到高峰,72小时后减少。Bcl-2表达至再灌注12小时达高峰,再灌注24~72小时组逐渐减弱。Bax表达至48小时达高峰,再灌注72小时减少。结论:Bcl-2于再灌注早期表达增强,Bax于再灌注中期表达增强,Bcl-2/Bax比例失衡可能是大鼠全脑缺血再灌注后神经细胞凋亡的机制之一。     

Effects of acupoint versus non-acupoint electroacupuncture on cerebral cortical neuronal Bcl-2, Bax and caspase-3 expression in a rat model of focal cerebral ischemia

BACKGROUND： Several studies have demonstrated that electroacupuncture by acupoint selection can inhibit cerebral cortical neuronal apoptosis following cerebral ischemia/reperfusion. OBJECTIVE： To validate the effects of electroacupuncture by acupoint selection on the expression level of cortical neuronal anti-apoptotic Bcl-2 protein and the apoptotic executive protein, caspase-3, in rat models of focal cerebral ischemia/reperfusion. DESIGN, TIME AND SETTING： This randomized grouping, neural cell and molecular biology animal experiment was performed at the Laboratory of Pharmacology of Traditional Chinese Medicine and the Laboratory Animal Center of Henan Institute of Traditional Chinese Medicine between November 2006 and May 2007. MATERIALS： Atotal of 40 healthy male adult Sprague-Dawley rats were randomly and evenly divided into four groups： sham-operated, model, electroacupuncture and non-aeupoint control. G6895 electro-acupuncture instruments were purchased from Shanghai Huayi Instrument Factory, China. Caspase-3, Bcl-2 and Bax kits were provided by Wuhan Boster Bioengineering Co., Ltd., China. METHODS： Middle cerebral artery occlusion was induced in the model, electroacupuncture and non-acupoint groups. In the electroacupuncture group, the acupoints Jianyu （LII5）, Waiguan （S J5）, Biguan （ST31）, and Zusanli （ST36） were given electroacupuncture. In the non-acupoint control group, at each time point （immediately after ischemia and after reperfusion, or 2 hours after reperfusion）, electroacupuncture was performed at the midpoints of Tianquan （PC2）-Quze （PC 3） line, Quze （PC 3）-Ximen （PC4） line, Zuwuli （LR10）-Yinbao （LR9） line, and Xiguan （LR7）-Zhongdu （LR6） line. Electroacupuncture parameters were set with a continuous wave with a frequency of 10 Hz, wave width 0.6 ms, voltage 1.5-3.0 V, and a duration of 10 minutes. The sham-operated and model groups received only animal fixation without electroacupuncture procedure. MAIN OUTCOME MEASURES： Five ra     

Effects of Rubus parvifolius L. on neuronal apoptosis and expression of apoptosis-related proteins in a rat model of focal cerebral ischemic-reperfusion injury

BACKGROUND:Studies have shown that the Ruhus parvifolius L.(RP)plant extract exhibits protective effects on cerebral ischemia.This effect is reflected in altered ischemic neuronal apoptosis and associated protein expression.OBJECTIVE:To explore the neuroprotective mechanism of RP after cerebral ischemia jniury.DESIGN,TIME AND SETTING:Randomized control experiment of cellular,molecular.and protein levels.The experiment was completed at Chongqing Medical University at the School of Pharmacy Laboratories and Basic Medical Institute from October 2005 to January 2006.MATERIALS:Twenty-four adult,male,Wistar rats,weighing(28±20)g.RP extract,which was a product of ethanol extraction.was provided by the LabOratory of Pharmaceutical Analysis,Chongqing Medical University.RP was dissolved in distilled water to a concentration of 10 mg/mL.All rats were randomly assigned into four groups:5 g/kg RP,10 g/kg RP,model,and sham-surgery,with 6 rats in each group.METHODS:In the 5 and 10 g/kg RP groups,as well as the model group,the middle cerebral artery was occluded(MCAO)for 60 minutes,resulting in focal cerebral ischemia,followed by reperfusion for 24 hours.Rats in the 5 and 10 g/kg RP groups received 5 and 10 g/kg RP,respectively.The RP treatment group received RP intragastrically(once a day)for 3 days.One hour after the last dose,rats were subjected to MCAO.The same surgical procedure was performed in the sham-surgery group,except the suture was introduced into the extemal carotid artery,but not advanced.Rats in the model group were subjected to MCAO.The sham-surgery and model groups received intragastrically administered normal saline once per day for 3 days.One hour after the last dose,the rats were subjected to surgery.MAIN OUTCOME MEASURES:TUNEL labeling and immunohistochemical methods were used to investigate changes in neuronal apoptosis and expression of the apoptosis-related proteins.Bax and Bcl-2.on the ischemic hemisphere,as well as the contralateral hemisphere,after administration of RP or normal saline.RESULTS:All 24 Wistar rats were included in the final analysis.without any loss.Compared with the sham-surgery group,the number of the TUNEL-positive cells in the model group,as well as the 5 and 10 g/kg RP treatment groups,were significantly increased in the ischemic hemisphere(P<0.05).Compared with the sham-surgery group,the number of Bcl-2-positive and Bax-positive cells increased significantly in the model group(P<0.01).The number of Bcl-2-positive cells increased,and the number of Bax-positive cells decreased in the model group,compared to the 5 and 10 g/kg RP treatment groups(P<0.05-0.01).CONCLUSION:RP can prevent neuronal apoptosis following cerebral ischemic-reperfusion of rats.The mechanism underlying the RP-induced neuroprotection in the cerebral ischemia iniury may be related to increased Bcl-2 expression and decreased Bax expression.     

bFGF对大鼠局灶性脑缺血后神经细胞凋亡及Bcl-2、Bax的影响

目的探讨bFGF对缺血后神经细胞的保护作用。方法用线栓法制作局灶性脑缺血大鼠模型,于术前1h、术后第1天、第2天连续3天侧脑室注射bFGF,分1μg／d、2μg／d、4μg／d3组,观察缺血程度、梗塞体积、Bcl-2、Bax蛋白的合成。结果;bFGF能减轻脑缺血程度,减少梗塞体积(25．2％)及凋亡细胞数,提高半暗带内Bcl-2蛋白的合成,减低缺血灶内Bax蛋白的合成,各剂量组间无显著差异。结论bFGF可作为一种有效的神经细胞保护剂,保护神经细胞免受缺血的损害。     

Angiopoietin-1 mRNA and Bcl-2 expression following estradiol treatment in ovariectomized rats with focal cerebral ischemia/reperfusion injury

BACKGROUND： Estrogen has been clinically demonstrated to attenuate ischemic brain injury. However, the precise mechanisms remain controversial. OBJECTIVE： To investigate the effects of estradiol on angiopoietin-1 mRNA and Bcl-2 expression, as well as apoptosis and cerebral blood flow, in ovadectomized rats with focal cerebral ischemia following reperfusion. DESIGN, TIME AND SETTING： Randomized, controlled, animal experiment. The study was performed at the Central Laboratory, Chongqing Medical University from September to December 2005. MATERIALS： Estradiol benzoate was purchased from Shanghai Ninth Pharmaceutical Factory; corn oil was purchased from Walmart Supercenter; TUNEL kit, rabbit anti-rat Bcl-2 polyclonal antibody, and biotin-labeled goat anti-rabbit antibody were purchased from Wuhan Boster, China. METHODS： Healthy, female, 6-month-old Wistar rats-wild-type and estrogen alpha receptor gene knockout （ERKO）-were randomly divided into estradiol and control groups with 25 animals in each group. The rats were intramuscularly injected with estradiol benzoate （100 μg/kg per day） at 30 days following bilateral ovariectomy or corn oil （1 mL/kg per day） for seven consecutive days. Following administration, cerebral ischemia/reperfusion models were established using the right middle cerebral artery occlusion （MCAO） method. After 30 minutes of MCAO, estradiol and control groups were separately injected with estradiol benzoate and corn oil with the above-mentioned doses. MAIN OUTCOME MEASURES： Cell apoptosis was determined by TUNEL; angiopoietin-1 mRNA and Bcl-2 gene expression was determined, respectively, by immunohistochemical staining and RT-PCR. In addition, changes in cerebral blood flow were measured by laser Doppler flowmetry. RESULTS： Changes in angiopoietin-1 mRNA and cerebral blood flow in estradiol-treated, wild-type, MCAO rats following ischemia/reperfusion were greater than in control rats （P 〈 0.01 or 0.05）. However, no significant difference was observed between estradiol-treated ERKO MCAO rats and control rats. In addition, estradiol-treated wild-type and ERKO MCAO rats exhibited significantly increased Bcl-2 expression （P 〈 0.05） and decreased number of apoptotic cells in brain tissues compared with control groups （P 〈 0.05）. CONCLUSION： Estradiol upregulated angiopoietin-1 mRNA and Bcl-2 expression, suggesting that estradiol might be involved in protective mechanisms of cerebral ischemia/reperfusion injury.     

bFGF对大鼠局灶性脑缺血后神经细胞凋亡及外Bcl-2、Bax的影响

目的：探讨bFGF对缺血后神经细胞的保护作用．方法：用线栓法制作局灶性日缺血大鼠模型,于术前1h、术后第1天、第 2天连续3天侧脑室注射bFGF,分 1μg／d、2μg／d、4 μg／d 3组,观察缺血程度、梗塞体积、Bcl-2、Bax蛋白的合成。结果：bFGF能减轻脑缺血程度,减少梗塞体积（25.2％）及凋亡细胞数,提高半暗带内Bcl-2蛋白的合成,减低缺血灶内Bax蛋白的合成,各剂量组间无显著差异。结论：bFGF可作为一种有效的神经细胞保护剂．保护神经细胞免受缺血的损害。     

Neuroprotective effects of total saponins from Rubus parvifolius L. on cerebral ischemia/ reperfusion injury in rats

This study examines the neuroprotective effects and mechanisms of action of total saponins from Rubus parvifolius L. (TSRP) on focal cerebral ischemia and reperfusion injury in rats. Focal cerebral ischemia and reperfusion injury was performed in rats using the suture method. The results indicate that intragastric injection of TSRP, at 5, 10 and 20 mg/kg, could decrease neurological impairment, reduce cerebral infarct volume, diminish pathological changes, and significantly inhibit the apoptosis of neurons surrounding the ischemic area. In addition, TSRP upregulated the expression of the anti-apoptotic factor Bcl-2, at the protein and mRNA levels, and it downregulated the expression of the pro-apoptotic factor Bax, at the protein and mRNA levels. These findings indicate that TSRP protects against cerebral ischemia/reperfusion injury, and that it may do so by regulating the expression of Bcl-2 and Bax.     

Influence of Tanshinone IIa on heat shock protein 70, Bcl-2 and Bax expression in rats with spinal ischemia/reperfusion injury

Tanshinone lla is an effective monomer component of Danshen, which is a traditional Chinese medicine for activating blood circulation to dissipate blood stasis. Tanshinone lla can effectively improve brain tissue ischemia/hypoxia injury. The present study established a rat model of spinal cord ischemia/reperfusion injury and intraperitoneally injected Tanshinone IIa, 0.5 hour prior to model establishment. Results showed that Tanshinone lla promoted heat shock protein 70 and Bcl-2 protein expression, but inhibited Bax protein expression in the injured spinal cord after ischemia/reperfusion injury. Furthermore, Nissl staining indicated a reduction in nerve cell apoptosis and fewer pathological lesions in the presence of Tanshinone lla, compared with positive control Danshen injection.