缺血/再灌注损伤,脑

摘要： 目的 探讨红景天苷联合亚低温对大鼠全脑缺血/再灌注(I/R)损伤后神经细胞形态和功能的影响及其作用机制.方法 选择雄性Wistar大鼠150只,按随机数字表法分为假手术组、模型组、单纯红景天苷治疗组、单纯亚低温治疗组、红景天苷联合亚低温治疗组,每组30只.单纯红景天苷治疗组和红景天苷联合亚低温治疗组制模前7d于同一时间腹腔注射红景天苷10 mg· kg-1·d“,最后一次给药30 min后建立全脑缺血模型;假手术组、模型组、单纯亚低温治疗组制模前7d同一时间腹腔注射等量生理盐水.采用改良Pusinelli四动脉阻断法建立全脑缺血模型,缺血10 min后实现再灌注;假手术组仅暴露双侧翼孔及游离颈总动脉,不凝闭椎动脉和夹闭颈总动脉.假手术组、模型组、单纯红景天苷治疗组保持肛温36.5°C;单纯亚低温治疗组和红景天苷联合亚低温治疗组大鼠迅速放入4°C环境使肛温降至32°C并维持2h.再灌注24 h后观察各组大鼠神经功能缺损程度评分(NDS)、脑组织含水量及海马区脑组织细胞形态、超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量.结果 与假手术组比较,模型组NDS、SOD均明显降低[NDS(分):50.70±1.64比76.70±1.34,SOD (U/mg):55.14±9.07比122.10±10.16],脑组织含水量、MDA均明显升高[脑含水量:(87.41±0.83)％比(78.30±0.75)％,MDA (nmol/mg):5.96±0.28比3.59±0.20,均P＜0.05].显微镜下观察到模型大鼠海马区细胞排列散乱,大量锥体细胞体积缩小,核固缩、深染及胞质嗜伊红,细胞间隙明显增大,胞质呈空泡样变,正常神经元数量显著减少;3个治疗组海马区细胞变性程度明显减轻.与模型组比较,单纯红景天苷治疗组、单纯亚低温治疗组、红景天苷联合亚低温治疗组NDS、SOD均明显升高(NDS分别为59.00±1.72、60.40±1.51、66.40±1.51,SOD分别为84.22±8.88、83.58±8.39、104.16±8.81),脑组织含水量、MDA均明显降低[脑组织含水量分别为(84.56±0.42)％、(83.94±0.36)％、(80.78±0.31)％,MDA分别为5.03±0.22、4.99±0.28、4.58±0.25,均P＜0.05],且以红景天苷联合亚低温治疗组变化程度较单纯红景天苷治疗组、单纯亚低温治疗组更显著[NDS(分):66.40±1.51比59.00±1.72、60.40±1.51,脑组织含水量:(80.78±0.31)％比(84.56±0.42)％、(83.94±0.36)％,SOD(U/mg):104.16±8.81比84.22±8.88、83.58±8.39,MDA (nmol/mg):4.58±0.25比5.03±0.22、4.99±0.28,均P＜0.05].结论 红景天苷和亚低温对大鼠全脑I/R损伤均有保护作用,且二者联合保护作用更佳,其机制可能与抗自由基损伤有关.

摘要： 目的 探讨腺苷A2a受体是否介导了人参皂苷Rb1对脑缺血/再灌注(I/R)损伤大鼠脑血流量的影响,为人参皂苷Rb1的脑保护机制提供新的理论基础.方法 将60只SD大鼠按随机数字表法分为生理盐水对照组、模型组、人参皂苷Rb1组、人参皂苷Rb1 +A2a受体拮抗剂组、A2a受体拮抗剂对照组5组,每组12只.采用大脑中动脉闭塞(MCAO)法建立大鼠脑I/R损伤模型.人参皂苷Rb1组在制模后即刻腹腔注射人参皂苷Rb1 (40 mg/kg);人参皂苷Rb1 +A2a受体拮抗剂组在制模前30 min腹腔注射A2a受体拮抗剂CSC(0.01 mg/kg),制模后即刻腹腔注射人参皂苷Rb1 (40 mg/kg);A2a受体拮抗剂对照组只在制模前30 min腹腔注射A2a受体拮抗剂CSC(0.01 mgkg);生理盐水对照组于制模后即刻腹腔注射等量生理盐水.每组6只大鼠于脑I/R损伤后24h进行行为学评分,并测量局部血流量,断头取脑组织测量脑梗死体积;另6只大鼠处死后取大脑皮质,测定丙二醛(MDA)、超氧化物歧化酶(SOD)含量.结果 与生理盐水对照组和模型组比较,人参皂苷Rb1组大鼠行为学评分明显降低[分:1(1～2)比3(2～4)、3(2～4),均P＜0.05],局部脑血流量明显增多(L/min:223.25±67.15比127.23±64.16、125.75±57.65,均P＜0.05),脑梗死体积明显减小[(24.73±8.29)％比(63.72±8.81)％、(65.13±7.92)％,均p＜0.05],MDA含量明显减少(U/mg:15.16±5.89比30.35±5.78、28.65±9.12,均P＜ 0.05),SOD活性明显增加(nmol/mg:125.19±12.39比92.42±9.82、89.59±10.85,均P＜0.05).与人参皂苷Rb1组相比,人参皂苷Rb1+A2a受体拮抗剂组大鼠行为学评分明显升高[分:3(2～4)比1(1～2),P＜0.05],局部脑血流量明显减少(L/min:181.35±61.33比223.25±67.15,P＜ 0.05),脑梗死体积明显增大[(40.25±9.14)％比(24.73±8.29)％,P＜0.05],MDA含量明显增加(U/mg:25.38±6.78比15.16±5.89,P＜0.05),SOD活性明显降低(nmol／mg:95.61±13.12比125.19±12.39,P＜ 0.05).A2a受体拮抗剂对照组各指标与模型组比较差异均无统计学意义.结论 腺苷A2a受体可能介导了人参皂苷Rb1对脑I/R损伤大鼠脑血流量的增加,为人参皂苷Rb1的脑保护机制提供了新的理论基础.

摘要： 目的：观察人参皂苷Rb1对缺血/再灌注（I/R）损伤大鼠脑血流量的影响，为探讨其脑保护机制提供新的理论基础。方法将24只大鼠按随机数字表法分为假手术组、模型组、生理盐水对照组、人参皂苷Rb1组，每组6只。采用大脑中动脉闭塞（MCAO）法复制大鼠脑I/R模型。人参皂苷Rb1组于制模后即刻腹腔注射人参皂苷Rb140 mg/kg，生理盐水对照组腹腔注射等体积生理盐水。脑I/R 24 h时监测大鼠局部脑血流量，观察大鼠行为学评分,并采用2，3，5-氯化三苯基四氮唑（TTC）染色法测量脑梗死体积。结果与假手术组比较，模型组梗死体积百分比〔（64.23±8.12）%比0%〕和神经行为评分〔分：3.0（2.0～4.0）比0（0～0），P＜0.05〕均明显升高，局部脑血流量明显减少（mL/min：125.75±57.65比225.01±78.25，P＜0.05）；与模型组和生理盐水对照组比较，人参皂苷Rb1组梗死体积百分比〔（23.62±8.74）%比（64.23±8.12）%、56.72±8.92〕和神经行为学评分〔分：0.5（0.0～2.0）比3.0（2.0～4.0）、3.5（1.0～4.0）〕均明显降低（P＜0.05），局部脑血流量明显增多（mL/min：177.25±75.36比125.75±57.65，132.65±58.65，P＜0.05）。结论人参皂苷Rb1可以增加I/R大鼠脑血流量，这可能是人参皂苷Rb1发挥脑保护作用的机制之一。%ObjectiveTo observe the effect of ginsenosides Rb1 on cerebral blood flow of rat models with cerebral ischemia/reperfusion (I/R) injury, which could provide a new theory of cerebral protective mechanism about ginsenosides Rb1.Methods Twenty-four rats were randomly divided into sham-operation group, model group, normal saline control group and ginsenosides Rb1 group, 6 rats in each group. The middle cerebral artery occlusion (MCAO) model was established by thread embolism method. At the end of I/R, in the rat of ginsenosides Rb1 group, ginsenosides Rb1 40 mg/kg was immediately intraperitoneally injected, while in the rat of normal saline control group, an equal volume of normal saline was injected intraperitoneally. After I/R for 24 hours, the cerebral local amount of blood flow was measured, the rats' behavior score was observed, and the volume of cerebral infarction was monitored by 2, 3, 5-triphenyl tetrazolium chloride (TTC) staining.Results The percentage of volume of cerebral infarction [(64.23±8.12)% vs. 0%] and behavior score [3.0 (2.0-4.0) vs. 0 (0-0),P< 0.05] in model group were significantly higher than those in sham-operation group, while the cerebral local amount of blood flow in model group was obviously lower than that in sham-operation group (mL/min: 125.75±57.65 vs. 225.01±78.25,P< 0.05); Compared with the model group and normal saline control group, the percentage of volume of cerebral infarction [(23.62±8.74)% vs. (64.23±8.12)%, 56.72±8.92] and behavior score [0.5 (0.0-2.0) vs. 3.0 (2.0-4.0), 3.5 (1.0-4.0)] in the ginsenosides Rb1 group were significantly lower, the cerebral local amount of blood flow was markedly increased in the ginsenosides Rb1 group (177.25±75.36 vs. 125.75±57.65, 132.65±58.65,P< 0.05).Conclusion Ginsenosides Rb1 can increase the cerebral blood flow in rats with cerebral I/R injury, which maybe one of the mechanisms of cerebral protection of Ginsenosides Rb1.

摘要： 目的 探讨高渗羟乙基淀粉200/0.5氯化钠注射液对脑缺血-再灌注大鼠颅内压和脑水肿的影响及可能机制.方法 采用随机对照动物实验研究方法,实验在中山大学实验动物中心进行.取28只雄性SD大鼠,随机(随机数字法)分为高渗羟乙基淀粉组、羟乙基淀粉组、对照组和假手术组.右侧大脑中动脉阻塞(middle cerebral artery occlusion,MCAO)法建立脑梗死再灌注大鼠模型,造模不成功另选大鼠再行手术补足手术组,于再灌注开始时尾静脉泵入高渗羟乙基淀粉200/0.5氯化钠注射液和羟乙基淀粉130/0.4氯化钠注射液,分别为206 mL/ (kg· d);造模后0、2、6、12、18、24 h时间点分别测量血浆胶体渗透压(plasma colloid osmotic pressure,COP)和颅内压(intracranial pressure,ICP);治疗24 h后测量右侧大脑半球含水量(brain water content,BWC).结果 高渗羟乙基淀粉组、羟乙基淀粉组和对照组术后各时间点ICP均显著高于假手术组;高渗羟乙基淀粉组术后ICP明显低于对照组和羟乙基淀粉组,但羟乙基淀粉组各时间点ICP和对照组比较差异无统计学意义.高渗羟乙基淀粉组和羟乙基淀粉组COP各时间点均显著高于对照组和假手术组,高渗羟乙基淀粉组和羟乙基淀粉组间差异均无统计学意义.高渗羟乙基淀粉组、羟乙基淀粉组和对照组脑含水量均显著高于假手术组[(81.24±0.36)％、(83.04±0.10)％、(83.14±0.41)％ vs.(78.37±0.37)％,P=0.000];高渗羟乙基淀粉组脑含水率显著低于对照组[(81.24±0.36)％ vs.(83.14±0.41)％,P=0.000)]和羟乙基淀粉组[(81.24±0.36)％ vs.(83.04±0.10)％,P=0.000];羟乙基淀粉组和对照组比较差异无统计学意义[(83.04±0.10)％vs.(83.14±0.41)％,P=0.578].结论 高渗羟乙基淀粉可明显改善脑缺血-再灌注大鼠急性期脑水肿、降低颅内压,但未能证实其提高COP对颅内压和脑水肿的影响.%Objective To investigate effects and its mechanisms of hypertonic saline hydroxyethyl starch 200/0.5 solution on intracranial pressure and brain water content in rats with ischemic cerebral edema.Methods All experiments were conducted in the animal experimental center of Sun Yat-sen University.The 28 male Sprague-Dawle (SD) rats were randomly (random number) divided into hypertonic saline hydroxyethyl starch group,hydroxyethyl starch group,control group and sham operation group,each n =7.Ischemic cerebral edema model was reproduced by middle cerebral artery occlusion (MCAO),followed by reperfusion after ischemia for 2 hours (If the moldel was not successful,other rats were operated to fill the missing models).Then reperfusion after ischemia 2 hours and received hypertonic saline hydroxyethyl starch and hydroxyethyl starch via tail vein at the beginning of reperfusion.The colloidal osmotic pressure (COP) and intracranial pressure (ICP) were evaluated on 0,2,6,12,18,24 hours after the surgery.The water content of the right hemisphere was measured on 24 h after the surgery.Results The ICP of hypertonic saline hydroxyethyl starch group,hydroxyethyl starch group and control group were significantly higher than that of sham operation group on 2,6,12,18,24 h after the surgery.The ICP of hypertonic saline hydroxyethyl starch group was significantly lower than those of hydroxyethyl starch group and control group on 2,6,12,18 and 24 h.But there was no significant difference in ICP of the hydroxyethyl starch group compared with that of control group at all time points.The COP of hypertonic saline hydroxyethyl starch group and hydroxyethyl starch group were significantly higher than the control group and sham operation group at each time point; There was no significant difference in COP (mmHg) of the hydroxyethyl starch group compared with that of hypertonic saline hydroxyethyl starch group at all time points.The brain water content (BWC) of hypertonic saline hydroxyethyl starch group,hydroxyethyl starch group and control group were significantly higher than that of sham operation group on 24 hours after the surgery [(81.24±0.36)％,(83.04±0.10)％,(83.14±0.41)％ vs.(78.37±0.37)％,all P=0.000],BWC of hypertonic saline hydroxyethyl starch group lower than these of hydroxyethyl starch group [(81.24±0.36)％ vs.(83.04 ±0.10) ％,P =0.000] and control group [(81.24 ±0.36)％ vs.(83.14 ±0.41) ％,P =0.000].There was no significant difference in BWC of the hydroxyethyl starch group compared with that of control group [(83.04 ± 0.10) ％ vs.(83.14 ± 0.41) ％,P =0.578].Conclusion Hypertonic saline hydroxyethyl starch solution could significantly ameliorate ischemic cerebral edema and reduce ICP,but the relationship between its elevated COP and reduced ICP has not been confirmed.

摘要： 目的 探讨异氟烷预处理或后处理对大鼠局灶性脑缺血/再灌注（I/R）时炎症因子、脂质过氧化等相关指标的影响.方法 将32只SD大鼠按随机数字表法分为对照组、模型组、异氟烷预处理组及异氟烷后处理组4组,每组8只.对照组不给予异氟烷与脑缺血处理;模型组大鼠接受90 min右侧大脑中动脉闭塞（MCAO）;异氟烷预处理组大鼠在MCAO前24 h给予2％异氟烷30 min;异氟烷后处理组大鼠在MCAO后,于再灌注开始时给予2％异氟烷60 min.操作结束24 h后取心脏血,检测血清炎症因子白细胞介素-1β（IL-1β）、肿瘤坏死因子-α（TNF-α）含量及脂质过氧化相关指标丙二醛（MDA）含量和超氧化物歧化酶（SOD）活性.采用逆转录-聚合酶链反应（RT-PCR）和蛋白质免疫印迹试验（Westen Blot）检测右侧脑组织基质金属蛋白酶（MMP-2、MMP-9）及紧密连接蛋白闭合蛋白5（Claudin-5）、咬合蛋白（Occludin）的mRNA和蛋白表达.结果 与对照组相比,模型组大鼠血清IL-1β（ng/L）、TNF-α（ng/L）及MDA（μmol/L）含量均显著增加,SOD（U/L）活性则显著下降（IL-1β：76.81±11.14比52.43±8.86,TNF-α：64.93±10.81比33.64±7.94,MDA：8.63±1.42比4.14±0.98,SOD：0.95±0.21比2.36±0.80,均P＜0.05）;与模型组比较,经异氟烷预处理或后处理后,大鼠血清IL-1β、TNF-α及MDA含量显著下降,SOD活性则显著回升（IL-1β：54.37±9.06、56.82±8.67比76.81±11.14,TNF-α：43.72±6.16、39.49±9.34比64.93±10.81,MDA：5.65 ±0.83、5.82±0.78比8.63±1.42,SOD：1.64±0.47、1.71±0.52比0.95±0.21,均P＜0.05）.同时,I/R可导致受损脑组织MMP表达升高,紧密连接蛋白表达降低;与模型组比较,经过异氟烷预处理或后处理后,大鼠受损脑组织中MMP-2与MMP-9的mRNA及蛋白表达均明显下降（MMP-2 mRNA：1.25±0.08、1.32±0.12比2.48±0.26,MMP-2蛋白：1.56±0.09、1.50±0.08比2.12±0.11; MMP-9 mRNA：1.26 ±0.13、1.20±0.12比2.74±0.28,MMP-9蛋白：1.53±0.04、1.51±0.05比2.23±0.09,均P＜0.05）,而Claudin-5和Occludin的mRNA及蛋白表达均显著回升（Claudin-5 mRNA：0.40±0.08、0.38±0.06比0.28±0.03,Claudin-5蛋白：0.80±0.06、0.81±0.07比0.39±0.02; Occludin mRNA：0.54±0.07、0.50±0.08比0.26±0.06,Occludin蛋白：0.64±0.06、0.69±0.05比0.49±0.02,均P＜0.05）.结论 异氟烷预处理或后处理均可缓解I/R引起的血清中炎症因子分泌及脂质过氧化程度,并可降低大脑组织中MMP对紧密连接蛋白的蛋白水解活性,减少紧密连接蛋白的缺失,从而减轻I/R损伤.

摘要： Objective To investigate the effects of preconditioning and postconditioning with isoflurane on pro-inflammatory cytokines and lipid peroxidation in focal cerebral ischemic/reperfusion (I/R) injury in rats.Methods Thirty-two Sprague-Dawley (SD) rats were randomly divided into four groups:control group,model group,isoflurane preconditioning group and isoflurane postconditioning group,with 8 rats in each group.Rats in control group did not receive any challenge.In rats of model group right middle cerebral artery occlusion (MCAO) was conducted for 90 minutes.Rats in isoflurane preconditioning group received 2％ isoflurane exposure for 30 minutes 24 hours before MCAO for 90 minutes.Rats in isoflurane postconditioning group were given 60-minute 2％ isoflurane exposure after reperfusion of right MCAO.Twenty-four hours after the procedure,all rats were anesthetized with isoflurane,and blood sample taken from the heart was centrifuged,and the pro-inflammatory cytokines,including interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α),and lipid peroxidation products such as malonaldehyde (MDA) and superoxide dismutase (SOD) were determined.The mRNA and protein expression levels of matrix metalloproteinase (MMP-2,MMP-9),tight junction protein Calaudin-5 and Occludin were determined by reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot.Results Compared with control group,serum levels of IL-1 β (ng/L),TNF-α (ng/L) and MDA (μmol/L) were elevated and activity of SOD (U/L) decreased in rats of model group (IL-1β:76.81 ± 11.14 vs.52.43 ± 8.86,TNF-α:64.93 ± 10.81 vs.33.64 ± 7.94,MDA:8.63 ± 1.42 vs.4.14 ± 0.98,SOD:0.95 ± 0.21 vs.2.36 ± 0.80,all P＜0.05).After isoflurane preconditioning and postconditioning,compared with model group,the levels of IL-1 β,TNF-α and MDA were lowered,while activity of SOD was increased (IL-1 β:54.37 ± 9.06,56.82 ± 8.67 vs.76.81 ± 1 1.14,TNF-α:43.72 ± 6.16,39.49 ± 9.34 vs.64.93 ± 10.81,MDA:5.65 ± 0.83,5.82 ± 0.78 vs.8.63 ± 1.42,SOD:1.64 ± 0.47,1.71 ± 0.52 vs.0.95 ± 0.21,all P＜0.05).Focal cerebral I/R injury could lead to an increased expression of MMP accompanied with a decreased expression of tight junction protein.Compared with model group,after isoflurane preconditioning and postconditioning,it was found that there were decreased mRNA and protein expression of MMP-2 and MMP-9 (MMP-2 mRNA:1.25 ± 0.08,1.32 ± 0.12 vs.2.48 ± 0.26,MMP-2 protein:1.56 ± 0.09,1.50 ± 0.08 vs.2.12 ± 0.11 ; MMP-9 mRNA:1.26 ± 0.13,1.20 ± 0.12 vs.2.74 ± 0.28,MMP-9 protein:1.53 ± 0.04,1.51 ± 0.05 vs.2.23 ± 0.09,all P＜0.05) and increased levels of Calaudin-5 and Occludin (Claudin-5 mRNA:0.40 ± 0.08,0.38 ± 0.06 vs.0.28 ± 0.03,Claudin-5 protein:0.80 ± 0.06,0.81 ± 0.07 vs.0.39 ± 0.02; Occludin mRNA:0.54 ± 0.07,0.50 ± 0.08 vs.0.26 ± 0.06,Occludin protein:0.64 ± 0.06,0.69 ± 0.05 vs.0.49 ± 0.02,all P＜0.05).Conclusion Preconditioning and postconditioning with isoflurane can lower the levels of pro-inflammatory cytokines and the degree of lipid peroxidation,and lower the hydrolytic activity of MMP to the tight junction protein in cerebral tissue,thereby decrease the loss of tight junction protein and alleviate I/R injury.%目的 探讨异氟烷预处理或后处理对大鼠局灶性脑缺血/再灌注(I/R)时炎症因子、脂质过氧化等相关指标的影响.方法 将32只SD大鼠按随机数字表法分为对照组、模型组、异氟烷预处理组及异氟烷后处理组4组,每组8只.对照组不给予异氟烷与脑缺血处理;模型组大鼠接受90 min右侧大脑中动脉闭塞(MCAO);异氟烷预处理组大鼠在MCAO前24 h给予2％异氟烷30 min;异氟烷后处理组大鼠在MCAO后,于再灌注开始时给予2％异氟烷60 min.操作结束24 h后取心脏血,检测血清炎症因子白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)含量及脂质过氧化相关指标丙二醛(MDA)含量和超氧化物歧化酶(SOD)活性.采用逆转录-聚合酶链反应(RT-PCR)和蛋白质免疫印迹试验(Westen Blot)检测右侧脑组织基质金属蛋白酶(MMP-2、MMP-9)及紧密连接蛋白闭合蛋白5(Claudin-5)、咬合蛋白(Occludin)的mRNA和蛋白表达.结果 与对照组相比,模型组大鼠血清IL-1β(ng/L)、TNF-α(ng/L)及MDA(μmol/L)含量均显著增加,SOD(U/L)活性则显著下降(IL-1β:76.81±11.14比52.43±8.86,TNF-α:64.93±10.81比33.64±7.94,MDA:8.63±1.42比4.14±0.98,SOD:0.95±0.21比2.36±0.80,均P＜0.05);与模型组比较,经异氟烷预处理或后处理后,大鼠血清IL-1β、TNF-α及MDA含量显著下降,SOD活性则显著回升(IL-1β:54.37±9.06、56.82±8.67比76.81±11.14,TNF-α:43.72±6.16、39.49±9.34比64.93±10.81,MDA:5.65 ±0.83、5.82±0.78比8.63±1.42,SOD:1.64±0.47、1.71±0.52比0.95±0.21,均P＜0.05).同时,I/R可导致受损脑组织MMP表达升高,紧密连接蛋白表达降低;与模型组比较,经过异氟烷预处理或后处理后,大鼠受损脑组织中MMP-2与MMP-9的mRNA及蛋白表达均明显下降(MMP-2 mRNA:1.25±0.08、1.32±0.12比2.48±0.26,MMP-2蛋白:1.56±0.09、1.50±0.08比2.12±0.11; MMP-9 mRNA:1.26 ±0.13、1.20±0.12比2.74±0.28,MMP-9蛋白:1.53±0.04、1.51±0.05比2.23±0.09,均P＜0.05),而Claudin-5和Occludin的mRNA及蛋白表达均显著回升(Claudin-5 mRNA:0.40±0.08、0.38±0.06比0.28±0.03,Claudin-5蛋白:0.80±0.06、0.81±0.07比0.39±0.02; Occludin mRNA:0.54±0.07、0.50±0.08比0.26±0.06,Occludin蛋白:0.64±0.06、0.69±0.05比0.49±0.02,均P＜0.05).结论 异氟烷预处理或后处理均可缓解I/R引起的血清中炎症因子分泌及脂质过氧化程度,并可降低大脑组织中MMP对紧密连接蛋白的蛋白水解活性,减少紧密连接蛋白的缺失,从而减轻I/R损伤.

摘要： Objective To investigate the protective effect of edaravone combined with propofol postconditioning against different periods of brain cortical cells injury in culture of suckling rat caused by ischemia/reperfusion(I/R)injury. Methods Cortical cells of Sprague-Dawley(SD)rats born within 24 hours had been cultured in vitro for 7 days. And then the cultured cells were randomly divided into blank control group,glutamate injury group,drug control group,and reperfusion of 30 minutes,2 hours and 12 hours of drug postconditioning groups. All drug postconditioning groups were injured by glutamate(200 μmol/L) for 30 minutes,2 hours and 12 hours. The nerve cells in each post-processing group were cultured in medium containing 100 μmol/L of edaravone and 3 mg/L of propofol after glutamate damage. The cortical cell survival rate,apoptosis rate,lactate dehydrogenase(LDH)leakage rate,activities of superoxide dismutase(SOD)and Na+-K+-ATPase,content of malondialdehyde(MDA)were also determined. The morphological changes of nerve cells were examined by inverted phase contrast microscope after staining with hematoxylin-eosin(HE). The ultrastructural changes of cells were observed by transmission electron microscope. Results Compared with those in the blank control group,in the glutamate injury group,the nerve cell survival rate〔(55.38±22.41)%vs.(87.04±23.95)%〕,the activity of SOD(kU/L:6.86±2.87 vs. 29.57±3.68),Na+-K+-ATPase activity (μmol·mg-1 ·h-1:0.76±0.73 vs. 2.39±0.90)were significantly decreased,and the rate of neuronal apoptosis〔(9.36±0.66)% vs.(6.06±0.20)%〕,LDH leakage rate〔(41.23±1.63)% vs.(36.82±4.63)%〕,the content of MDA(μmol/L:0.515±0.102 vs. 0.294±0.105) were significantly increased(P<0.05 or P<0.01). With the postconditioning and its prolongation,the nerve cell survival rate,the activities of SOD and Na+-K+-ATPase were gradually decreased in all drug postconditioning groups,while the apoptosis rate,LDH leakage rate and MDA content were gradually increased. Compared with those in glutamate injury group,the nerve cell survival rate〔(88.08±14.25)%,(83.33±22.90)%〕,the apoptosis rate〔(5.96±0.28)%,(7.25±0.34)%〕,LDH leakage rate〔(36.07±2.19)%,(38.04±1.44)%〕,SOD activity(31.00±5.84,18.22±6.19),Na+-K+-ATPase activity(2.50±0.57,1.59±0.64)and the content of MDA(0.270±0.146,0.343±0.091)in drug control group and 30-minute reperfusion of drug postconditioning group were improved,with statistical differences(P<0.05 or P<0.01). Compared with glutamate injury group and other drug postconditioning groups,the morphological changes of nerve cells damage in the 30-minute reperfusion of drug postconditioning group was lighter under inverted phase contrast microscope. Conclusion The protective effect of postconditioning with edaravone(100 μmol/L)combined with propofol(3 mg/L)in the 30-minute reperfusion after neonatal cortex cells with I/R injury was very remarkable.%　　目的研究不同时间窗联合应用依达拉奉及异丙酚后处理对乳鼠离体脑皮质细胞缺血/再灌注(I/R)损伤的保护作用.方法体外培养出生24 h内SD大鼠脑皮质细胞7 d,按随机数字表法分为空白对照组、谷氨酸损伤组、药物联合对照组及再灌注30 min、2 h、12 h后处理组.各药物后处理组分别于谷氨酸(200μmol/L)损伤后30 min、2 h、12 h 用含100μmol/L依达拉奉及3 mg/L异丙酚的培养基联合后处理原代培养的神经细胞.测定神经细胞存活率、细胞凋亡率、乳酸脱氢酶(LDH)漏出率、超氧化物歧化酶(SOD)活性、Na+-K+-ATP酶活性、丙二醛(MDA)含量；用苏木素-伊红(HE)染色后倒置相差显微镜下观察细胞形态,透射电镜下观察细胞超微结构变化.结果与空白对照组比较,谷氨酸损伤组神经细胞存活率〔(55.38±22.41)%比(87.04±23.95)%〕、SOD活性(kU/L：6.86±2.87比29.57±3.68)、Na+-K+-ATP酶活性(μmol·mg-1·h-1：0.76±0.73比2.39±0.90)均显著下降,神经细胞凋亡率〔(9.36±0.66)%比(6.06±0.20)%〕、LDH漏出率〔(41.23±1.63)%比(36.82±4.63)%〕、MDA含量(μmol/L：0.515±0.102比0.294±0.105)均明显升高(P<0.05或P<0.01).随着处理和后处理时间延长,药物联合各组细胞存活率、SOD活性、Na+-K+-ATP酶活性逐渐下降,细胞凋亡率、LDH漏出率、MDA含量逐渐升高；与谷氨酸损伤组比较,药物联合对照组和药物联合再灌注30 min后处理组细胞存活率〔(88.08±14.25)%、(83.33±22.90)%〕、细胞凋亡率〔(5.96±0.28)%、(7.25±0.34)%〕、LDH漏出率〔(36.07±2.19)%、(38.04±1.44)%〕、SOD活性(31.00±5.84、18.22±6.19)、Na+-K+-ATP酶活性(2.50±0.57、1.59±0.64)、MDA含量(0.270±0.146、0.343±0.091)均明显改善(P<0.05或P<0.01).镜下可见药物联合再灌注30 min后处理组细胞形态受损较谷氨酸损伤组及其他后处理组轻.结论在乳鼠离体脑皮质细胞谷氨酸损伤后30 min内予以100μmol/L依达拉奉及3 mg/L异丙酚两种药物联合处理对I/R损伤脑细胞具有明显保护作用.

摘要： 目的探讨青蒿素对脑缺血/再灌注(I/R)损伤大鼠肿瘤坏死因子-α(TNF-α)的影响.方法将40只Wistar大鼠按随机数字表法分为假手术组、模型组及青蒿素低、中、高浓度组,每组8只.采用大脑中动脉闭塞法(MCAO)复制局灶性脑I/R损伤大鼠模型.制模成功后,假手术组和模型组均灌胃30 ml生理盐水,青蒿素低、中、高浓度组分别由腹腔注射青蒿素200、300、400 mg/kg,均每日1次,连续60 d.采用放射免疫法(RIA)检测血清TNF-α含量,逆转录-聚合酶链反应(RT-PCR)和蛋白质免疫印迹法(Western blotting)检测关节软骨组织TNF-αmRNA和蛋白表达.结果与假手术组比较,模型组和青蒿素各浓度组血清TNF-α含量和组织TNF-αmRNA及蛋白表达均明显升高；与模型组比较,青蒿素各浓度组血清TNF-α含量及组织TNF-αmRNA和蛋白表达均明显下降(均P<0.05),以青蒿素高浓度组下降更显著〔血清TNF-α(μg/L)：51.9±9.4比88.3±9.7,TNF-αmRNA：0.20±0.14比0.77±0.31,TNF-α蛋白：0.19±0.08比0.76±0.09,均P<0.05〕.结论青蒿素可降低脑I/R损伤大鼠TNF-α的表达,有效控制炎症反应进程.%Objective To detect the effect of Artemisia annua on tumor necrosis factor-α(TNF-α)in rats with acute cerebral ischemia/reperfusion(I/R)injury. Methods Forty Wistar rats were randomized into sham operation group,model group,Artemisia annua high-,middle-and low-dose groups,each n=8. The model of focal cerebral I/R injury was reproduced by middle cerebral artery occlusion(MCAO). In the sham operation group and model group,30 ml normal saline was administered intra-gastrically,while in Artemisia annua low-,middle-and high-dose groups,200,300 and 400 mg/kg Artemisia annua were intra-peritoneally injected,respectively,once a day for consecutive 60 days in all the groups. The serum content of TNF-αwas detected by radioimmunoassay,and reverse transcription-polymerase chain reaction(RT-PCR)and Western blotting methods were used to determine the expressions of tissue TNF-α mRNA and protein. Results Compared with the sham operation group,the concentration of TNF-α in serum,and the expressions of tissue TNF-α mRNA and protein in model group and various Artemisia annua groups were significantly increased;compared with the model group,the concentration of TNF-αin serum,and the expressions of tissue TNF-αmRNA and protein in various Artemisia annua groups were significantly decreased(all P<0.05),the greatest descent being in the high-dose group〔TNF-αin serum(μg/L):51.9±9.4 vs. 88.3±9.7,TNF-αmRNA:0.20±0.14 vs. 0.77±0.31,TNF-αprotein:0.19±0.08 vs. 0.76±0.09, all P<0.05〕. Conclusion Artemisia annua can down-regulate the expression of TNF-αand effectively inhibit the process of inflammation in rates with acute cerebral I/R injury.

摘要： Objective To investigate the protective effect of combined pretreatment of edaravone and propofol on cerebral cortex with ischemia/reperfusion ( I/R ) injury and its therapeutic window.Methods Sprague Dawley (SD) rat brain cortex cells harvested within 24 hours of birth were cultured in vitro for 7 days.The cells were then divided into blank control group,glutamate injury group,24-hour drug precondition control group,and 24-,2-,0-hour drug precondition groups according to random number table.The nerve cells in each pretreatment group were cultured in medium containing 100 μmol/L of edaravone and 3 mg/L of propofol 24,2,or 0 hour before glutamate damage (200 μmol/L for 0.5 hour).Nerve cell survival or damage was determined by methyl thiazolyl tetrazolium (MTT),lactate dehydrogenase (LDH) leakage rate,and nerve cell Na+-K+-ATPase activity.The oxidation and anti-oxidation ability of nerve cells was observed by determining superoxide dismutase (SOD) activity (xanthine oxidase),malondialdehyde (MDA) content (thiobarbituric acid).Nerve apoptosis was detected by flow cytometry.Results Compared with blank control group,in the glutamate injury group,nerve cell survival rate [ (62.2 ± 23.4)％ vs.(90.5 ± 14.8)％],the activity of SOD (U/ml:6.864 ± 2.872 vs.29.569 ± 3.684),Na+-K+-ATPase activity ( U· mg-1· h-1:0.318 ± 0.146 vs.0.636 ± 0.168 ) were significantly decreased,and rate of neuronal apoptosis [ (9.4 ±0.7)％ vs. (6.1 ±0.2)％ ],the content of MDA (nmol/ml:0.515 ±0.101 vs.0.294 ±0.105),LDH leakage rate [(41.2 ± 1.6)％ vs.(36.8 ±4.6)％]were significantly increased (P＜0.05 or P＜0.01 ).Compared with glutamate injury group,the cell survival rate and the activity of SOD and Na+-K+-ATPase were significantly increased in the drug pretreatment groups,and apoptosis rate,MDA content,and LDH leakage rate were significantly decreased with time-department,and effect in the 24-hour pretreatment group was most significant [survival rate of cell: (89.2 ±30.3)％vs. (62.2±23.4)％,SOD activity (U/ml):17.780±4.514 vs.6.864±2.872,Na+-K+-ATPase activity ( U· mg-1· h-1 ):0.541 ± 0.052 vs.0.318 ± 0.146,the rate of cell apoptosis:( 6.7 ± 0.4 )％ vs.(9.4 ± 0.7 ) ％,the content of MDA (nmol/ml):0.319±0.101 vs.0.515±0.101,LDHleakagerate: (37.2±1.4)％vs.(41.2±1.6)％,all P＜0.01].Conclusion The synergistic protective effect of pretreatment with edaravone combined with propofol on neonatal rat brain cortex cells with I/R injury in vitro was evident; and 24-hour pretreatment is the best time window of protection for the cerebral neurons.%目的 观察依达拉奉联合异丙酚预处理对乳鼠离体脑皮质细胞缺血/再灌注(VR)损伤的保护作用以及脑保护效应的时间窗.方法 体外培养出生24 h内SD乳鼠脑皮质细胞7d,按随机数字表法分为空白对照组、浴氨酸损伤组、药物预处理24 h对照组及药物预处理24、2、0h组.各药物预处理组分别于谷氨酸损伤(200 μmol/L 0.5 h)前24、2、0h用含100 μmol/L依达拉奉和3 mg/L异丙酚的培养基联合预处理原代培养的神经细胞.通过测定神经细胞存活率[四甲基偶氮唑盐(MTT)比色法]、乳酸脱氢酶(LDH)漏出率、神经细胞Na+-K+-ATP酶活性观察神经细胞成活和细胞损伤情况；通过测定超氧化物歧化酶(SOD)活性(黄嘌呤氧化酶法)、丙二醛(MDA)含量(硫代巴比妥酸法)观察神经细胞抗氧化和氧化水平；流式细胞仪检测神经细胞凋亡情况.结果 与空白对照组比较,谷氨酸损伤组神经细胞存活率[(62.2±23.4)％比(90.5±14.8)％]、SOD活性(U/ml:6.864±2.872比29.569±3.684)、Na+-K+-ATP酶活性(U·mg4·h-1:0.318±0.146比0.636±0.168)均显著下降,神经细胞凋亡率[(9.4±0.7)％比(6.1±0.2)％]、MDA含量(nmol/ml:0.515±0.101比0.294±0.105)、LDH漏出率[(41.2±1.6)％比(36.8±4.6)％]均显著升高(P＜0.05或P＜0.01).与谷氨酸损伤组比较,药物预处理组细胞存活率、SOD活性、Na+-K+-ATP酶活性均显著升高,细胞凋亡率、MDA含量、LDH漏出率均显著下降,并呈时间依赖性,以预处理24 h组作用最为显著[细胞存活率:(89.2±30.3)％比(62.2±23.4)％,SOD活性(U/ml):17.780±4.514比6.864±2.872,Na+-K+-ATP酶活性(U·mg-1·h-1):0.541±0.052比0.318±0.146,细胞凋亡率:(6.7±0.4)％比(9.4±0.7)％,MDA含量(nmol/ml):0.319±0.101比0.515±0.101,LDH漏出率:(37.2±1.4)％比(41.2±1.6)％,均P＜0.01].结论 依达拉奉联合异丙酚预处理对乳鼠离体脑皮质细胞I/R损伤有明显协同保护作用；且以24 h作为时间窗的脑保护作用最明显.

摘要： 目的 探讨羟乙基淀粉对脑缺血/再灌注(I/R)大鼠血浆胶体渗透压(COP)的调节作用及其对颅内压(ICP)的影响.方法 采用随机对照动物实验研究方法,将24只雄性SD大鼠分为假手术组、模型组和羟乙基淀粉组,每组8只.采用大脑中动脉闭塞法(MCAO)建立脑I/R动物模型,于缺血2 h后再灌注,再灌注开始时尾静脉泵入羟乙基淀粉130/0.4氯化钠注射液206 ml·kg-1·d-1.于术后0、2、6、12、18、24 h分别测定ICP和COP;术后24 h测量右侧大脑半球含水量,用免疫组化法观察神经元凋亡情况.结果 模型组和羟乙基淀粉组术后2 h ICP(mm Hg,1 mm Hg=0.133 kPa)即显著高于假手术组(11.50±1.43、12.48±0.75比7.95±0.92,均P＜0.05),之后随时间延长,两组ICP逐渐升高,至24 h达峰值(22.76±0.72、23.32±0.98比8.15±1.09,均P＜0.05);但羟乙基淀粉组与模型组各时间点比较均无差异.羟乙基淀粉组术后各时间点COP(mm Hg)显著高于模型组和假手术组,于术后6 h达高峰(13.49±0.50比12.04±0.47、12.00±0.39,均P＜0.01);模型组与假手术组各时间点比较均无差异.羟乙基淀粉组和模型组脑组织含水量、神经元凋亡率均显著高于假手术组[脑组织含水量:(80.16±0.44)%、(80.59±0.67)%比(78.72±0.52)%;神经元凋亡率:(44.27±7.86)%、(42.82±7.82)%比(3.26±0.00)%,P＜0.05或P＜0.01];但羟乙基淀粉组与模型组比较差异无统计学意义(均P＞0.05).结论 羟乙基淀粉可以有效提高COP,但尚不能证实提高COP是否能够降低ICP、减轻脑水肿.%Objective To investigate the regulatory effect of hydroxyethyl starch on colloidal osmotic pressure(COP), and its effect on intracranial pressure(ICP)in rats with cerebral ischemia/reperfusion (I/R)injury. Methods Twenty-four male Sprague-Dawley(SD)rats were randomly divided into sham operation group, model group and the hydroxyethyl starch group, each n = 8. Cerebral I/R model was reproduced by middle cerebral artery occlusion(MCAO), followed by reperfusion after ischemia for 2 hours.Rats in hydroxyethyl starch group received hydroxyethyl starch 130/0. 4 206 ml · kg-1 · d-1 via tail vein at the beginning of reperfusion. ICP and COP were evaluated at 0, 2, 6, 12, 18, 24 hours after the surgery.The rats were sacrificed by decapitation. The water content of the right hemisphere was measured at 24 hours after the surgery, and the ratio of apoptosis of neurons was observed by immunohistochemical method. Results Two hours after surgery the ICP(mm Hg, 1 mm Hg=0.133 kPa)of model group and hydroxyethyl starch group was significantly increased compared with sham operation group(11. 50±1.43,12. 48±0. 75 vs. 7. 95±0. 92, both P＜0. 05). With prolongation of time, the ICP gradually increased and reached the peak at 24 hours(22. 76±0. 72, 23. 32±0. 98 vs. 8. 15±1.09, both P＜0. 05). But there was no significant difference in ICP in the hydroxyethyl starch group compared with that of the model group at all time points. The COP(mm Hg)of hydroxyethyl starch group was significantly higher than the model group and sham operation group at each time point, and peaked at 6 hours after surgery(13. 49±0. 50 vs. 12.04±0. 47, 12. 00±0. 39, both P＜0. 01). There was no significant difference in COP between the model group and the sham operation group at all time points. The brain water content, neuronal apoptosis of hydroxyethyl starch group and model group was significantly higher than sham operation group[brain water content:(80. 16 ± 0. 44)%,(80. 59 ± 0. 67)% vs.(78. 72 ± 0. 52)%; neuronal apoptosis:(44. 27 ± 7. 86)%,(42. 82 ± 7.82)% vs.(3. 26 ± 0. 00)%, P ＜ 0. 05 or P ＜ 0. 01], but there was no significant difference between the hydroxyethyl starch group and model group(both P＞0. 05). Conclusion Intravenous injection of hydroxyethyl starch 130/0.4 can increase the plasma COP, but it can not significantly reduce ICP and brain water content, and it also can not improve the neuronal apoptosis.

摘要： 本发明提供了一种口服富勒烯材料在制备预防和/或治疗心肌缺血再灌注损伤或缺血性心脏病药物中的应用。所述应用是富勒烯材料在制备预防和/或治疗如下1)‑7)中的至少一种疾病或病况的药物或药物载体中的应用：1)预防和/或治疗心肌缺血再灌注损伤或缺血性心脏病；2)保护心肌细胞；3)抑制细胞的凋亡；4)促进细胞的氧化损伤修复；5)改善心脏的结构和功能；6)缓解心肌缺血带来的心肌水肿、出血、超微结构改变等症状；7)改善心肌缺血引起的远端组织损伤。

摘要： 本发明公开了一种全肝缺血诱发缺血再灌注损伤的动物模型的构建方法。本发明所述动物模型设计合理、操作简单、重复性好，符合大鼠原味肝移植无肝期血流改变，术后存活率高。同时在结束后经病理分析，可导致肝外多器官(心脏、胰腺、结肠、小肠、肾脏)损伤。能有效解决现有研究目标必须完整复制肝移植过程，时间久、死亡率高、人员技术要求高等问题。同时可延伸到其他需阻断肝脏血流造成肝外器官缺血再灌注损伤的基础研究。

摘要： 在一些实施方案中，本发明涉及包括由结构式(I)表示的一种基本上纯的化合物的组合物：

摘要： 本发明涉及生物医药领域，尤其是一种脑缺血再灌注损伤生物标志物及养阴通脑颗粒的应用。一种脑缺血再灌注损伤的生物标志物，由基因标志物和代谢标志物组成。本研究中，我们发现缺血侧脑组织中7种蛋白质和对应代谢物的调控水平受到影响，并且这些异常明显通过YYTN治疗得到改善。包括Acadl、Gmps、Guk1、Ak2、Maob、Glb1、Hexb，其中Acadl、Gmps、Maob、Glb1、Hexb是枢纽基因，受影响的途径是饱和脂肪酸β氧化、嘌呤代谢、组氨酸代谢以及鞘糖脂生物合成‑神经节苷脂系列。

摘要： 本发明提供了一种口服富勒烯材料在制备预防和/或治疗心肌缺血再灌注损伤或缺血性心脏病药物中的应用。所述应用是富勒烯材料在制备预防和/或治疗如下1)‑7)中的至少一种疾病或病况的药物或药物载体中的应用：1)预防和/或治疗心肌缺血再灌注损伤或缺血性心脏病；2)保护心肌细胞；3)抑制细胞的凋亡；4)促进细胞的氧化损伤修复；5)改善心脏的结构和功能；6)缓解心肌缺血带来的心肌水肿、出血、超微结构改变等症状；7)改善心肌缺血引起的远端组织损伤。

摘要： 本发明公开了苯扎明在制备防治心肌缺血再灌注损伤或缺血性心脏病药物中的应用。通过对急性冠脉综合征患者外周血进行转录组数据分析，基于共表达通路和药物基因集富集分析方法，得到具有防治急性冠脉综合征的候选药物列表，发现其中包括若干具有急性冠脉综合征上市药物，之前未被发现有防治急性冠脉综合征作用的苯扎明位列其中，并在模拟心肌缺血再灌注损伤和缺血性心脏病的缺氧复氧细胞模型上验证了其效果与常见β受体阻滞剂心血管疾病药物美托洛尔的作用相当，因此，苯扎明可以应用于制备防治心肌缺血再灌注损伤或缺血性心脏病药物。

摘要： 本发明公开了一种全肝缺血诱发缺血再灌注损伤的动物模型的构建方法。本发明所述动物模型设计合理、操作简单、重复性好，符合大鼠原味肝移植无肝期血流改变，术后存活率高。同时在术后经病理分析，可导致肝外多器官(心脏、胰腺、结肠、小肠、肾脏)损伤。能有效解决现有研究目标必须完整复制肝移植过程，时间久、死亡率高、人员技术要求高等问题。同时可延伸到其他需阻断肝脏血流造成肝外器官缺血再灌注损伤的基础研究。

摘要： 本发明提供了一种减少缺血再灌注对心肌造成的损伤的药物或治疗心脏缺血性疾病的药物，它是以DNA四面体为活性成分，加上药学上可接受的辅料制备而成；所述DNA四面体是由4条DNA单链以相同物质的量，杂交形成的四面体结构。本发明的药物可以对心肌有良好的保护和修复功能，并抑制心肌细胞的凋亡，对心肌梗死等缺心脏缺血性疾病有良好的疗效。

摘要： 本发明公开了一种在衰老心肌缺血后处理缺血再灌注损伤治疗用miRNA标志物及其应用，属于分子生物学和基因工程技术领域，测血液样本中miR‑30a表达水平的试剂在制备治疗在衰老心肌缺血后处理缺血再灌注损伤产品中的应用，用于检测人血液样本的和用于检测小鼠或大鼠心肌样本中的所述miR‑30a的序列如SEQ：ID：NO：1所示。本发明首次提供一种用于IPostC预防或治疗I/R损伤的稳定性很好的药物组合物，为研制抗I/R损伤所致的心肌梗死的新药开辟新的途径。

摘要： 本发明公开了一种miR‑376a‑5p在制备治疗急性脑缺血和脑缺血再灌注损伤药物中的应用，所述miR‑376a‑5p具有如SEQ ID No:1和SEQ ID No:2所示的碱基序列。本发明发现miR‑376a‑5p是急性缺血性卒中神经保护的潜在靶点，上调miR‑376a‑5p是改善急性脑缺血血管再通治疗效果的一种神经保护新策略，本发明为miR‑376a‑5p治疗急性脑缺血和脑缺血再灌注损伤提供了新的方法，将成为治疗急性缺血性卒中血管内介入再通后再灌注损伤的创新性手段。