Effects of L-Tetrahydropalmatine on Energy Metabolism,Endothelin-1 and NO during Acute Cerebral Ischemia-reperfusion of Rats

To investigate the effects of L-tetrahydropalmatine (L-THP) on ener-gy metabolism, endothelin-1 (ET-1 ) and NO during acute cerebral ischemia-reperfusion of rats, 24 Wistar rats were randomly divided into four groups, with 6rats in each group: sham-operation group, simple ischemia group, ischemia-reperfusion group and treatment group (L-THP group). Cerebral ATP, lactate,ET-1 and NO levels were measured in all groups. Our results showed that treat-ment with L-THP could increase cerebral ATP levels, but decrease cerebral lac-tate, ET-1 and NO concentrations during ischemia-reperfusion in the treatmentgroup. It is concluded that L-THP could improve cerebral energy metabolism and protect the injured brain tissue, the mechanism of which might be related to suppression of overproduction of ET-1 and NO.

Experimental Study on the Protection of Agrimony Extracts from Different Extracting Methods against Cerebral Ischemia-Reperfusion Injury

Objective To study the protective effect of agrimony extracts from different extracting methods on cerebral ischemia-reperfusion injury in rats, in order to optimize the extraction scheme of agrimony.Methods Male rats were randomly assigned into seven groups: 1. Sham-operated group, 2. Untreated MCAO group (MCAO), 3. Petroleum ether extract of Agrimonia pilosa treated MCAO group (PEA), 4. Ethyl acetate extract of Agrimonia pilosa treated MCAO group (EAEA), 5. Ethanol extract of Agrimonia pilosa treated MCAO group (EEA), 6. Water extract of Agrimonia pilosa treated MCAO group (WEA), 7. Nimodipine treated MCAO group (NP). Intragastrical drug administration (i.g) was performed at 0 and 6 hours after MCAO.Neurological function tests were performed after reperfusion for 24 hours, then the brain was removed for the evaluations of the cerebral infarction volume (percentage of total brain volume) by immunohistochemistry,histological changes (hematoxylin-eosin staining), Na+/K+-ATPase, Ca2+-ATPase (modified method of Svoboda and Mosinger), mRNA expression of Tumor suppressor gene (P53) and hot shock protein (HSP70)(quantitative real-time PCR).Results The neurological function of MCAO group had significantly higher scores than the sham group (P<0.01). The WEA group showed a significantly lower neurological score than the MCAO group (P<0.05),indicating the protective effect of WEA on neurological deficits. The mean infarction volumes of WEA (13.5±6.6%, F=4.75, P<0.01), EEA (19.90±6.90%, F=5.23, P<0.01), PEA (20.40±5.30%, F=4.68,P<0.01) and EAEA (22.50±10.50%, F=6.25, P<0.05) group were all significantly smaller than that of MCAO group (29.40±6.50%). HE staining demonstrated that, compared to the treated groups, the infarcted cerebral tissue of MCAO group had more swelling neural cells, lighter stained nucleus, fewer and irregularly distributed neurons. The activity of Na+/K+-ATPase and Ca2+-ATPase reduced in the MCAO group (3.67±0.48 U/mg,1.28±0.26 U/mg, respectively), and were significantly higher in WEA group (7.56±0.85 U/mg, F=12.65,P=0.010; 3.59±0.22 U/mg, F=8.32, P=0.041, respectively). The MCAO group showed significantly elevated P53 and HSP70 mRNA expressions compared to the sham group (P<0.01, P<0.05). P53 mRNA expressions in Agrimony extracts treated groups were significantly lower than that of the MCAO group (all P<0.01), with the WEA group showing the greatest difference from MCAO group. The HSP70 mRNA level of the treated groups were not significantly different from that of the MCAO group.Conclusions Treatment using water extracts of agrimony can promote the best functional and metabolic recovery for rat model of cerebral ischemia-reperfusion injury, which maybe relate with the upregulation of energy metabolism in nerve cells after MCAO.

Effects of Electroacupuncture of Different Intensities on Energy Metabolism of Mitochondria of Brain Cells in Rats with Cerebral Ischemia-Reperfusion Injury

Objective： To observe the effects of electroacupuncture （EA） of different intensities on lactate dehydrogernase （LDH）, succinate dehydrogenase （SDH） and ATPase in brain tissue of rats with cerebral ischemia-reperfusion injury （CI/R）. Methods： Forty male SD rats were uniformly randomized into sham operation group （group A）, CI/R group （group B）, CI/R＋5 mA EA （group C）, CI/R＋3 mA EA （group D） and CI/R＋I mA EA （group E） groups with eight rats in each group. Transient general brain ischemia was induced by four-vessel occlusion and reperfusion. The rats in group C, group D and group E were punctured and stimulated at Baihui （GV20）, Mingmen （GV4） and Zusanli （ST36） with the same intermittent and rarefaction-dense wave （30 to 50 Hz） and different electric current intensities： 5 mA, 3 mA and 1 mA for 20 min after CI/R. Then the activities of Na^＋- ^＋-ATPase, SDH and LDH in mitochondria of brain tissue were measured by spectrophotometry. The ischemic cerebral cortex tissue was taken for observing the ultrastructure changes of impaired nerve cells. Results： Compared with group A, the activities of LDH, SDH and Na^＋-K^＋-ATPase were Iowerer in the group B （P〈0.05 or P〈0.01）. However, the activities of LDH, SDH and Na^＋-K^＋-ATPase were higher in the group D than those in the group B （P〈0.05 or P〈0.01）. In group A, the anatomical structure of the cerebral cortex cells was basically normal; in group B, the neuronal cellular structures were severely damaged, the neuronal mitochondria got swelling, the mitochondrial cristae were broken, the medullated nerve fibers were not integrated. In group C, group D and group E, the ultrastructure of impaired neuron were improved. Group D was the best among three groups above. Conclusion： EA of 3 mA intensity could strengthen aerobic metabolism by elevating the activities of SDH and LDH, meanwhile maintaining the ionic equilibrium in the exterior and interior brain cell and relieving the cellular edema bv reinforcina the activities of Na^＋-K^＋-ATPase.

Combination of Total Astragalus Extract and Total Panax Notoginseng Saponins Strengthened the Protective Effects on Brain Damage through Improving Energy Metabolism and Inhibiting Apoptosis after Cerebral Ischemia-Reperfusion in Mice

Objective： To explore the effects and molecular mechanisms of the combination between total Astragalus extract （TAE） and total Panax notoginseng saponins （TPNS） against cerebral ischemia- reperfusion injury. Methods： C57BL/6 mice were randomly divided into sham-operated group, model group, TAE （110 mg/kg） group, TPNS （115 mg/kg） group, TAE-TPNS combination group and Edaravone （4 mg/kg） group, treated for 4 days, then, cerebral ischemia-repeffusion injury was established by bilateral common carotid artery （CCA） ligation for 20 min followed by reperfusion for 1 and 24 h. Results： TPNS could increase adenosine triphosphate （ATP） level, TAE and TAE-TPNS combination increased ATP, adenosine diphosphate （ADP） contents and Na＋-K＋-ATPase activity, and the effects of TAE-TPNS combination were stronger than those of TAE or TPNS alone after reperfusion for 1 h. After reperfusion for 24 h, TAE, TPNS and TAE-TPNS combination significantly increased neurocyte survival rate and decreased the apoptosis rate as well as down-regulated the expression of phosphorylated c-June N-terminal kinasel/2 （p-JNK1/2）, cytochrome C （Cyt C）, cysteine aspartic acid-specific protease （Caspase）-9 and Caspase-3. Furthermore, the effects in TAE-TPNS combination were better than those in TAE or TPNS alone. Conclusion： The combination of TAE 110 mg/kg and TPNS 115 mg/kg could strengthen protective effects on cerebral ischemia injury, the mechanism underlying might be related to improving jointly the early energy metabolism, and relieving the delayed apoptosis via inhibiting the mitochondrial apoptosis pathway of JNK signal transduction.

活血荣络方对脑缺血再灌注损伤大鼠SIRT1/PGC-1α信号通路的影响

目的观察活血荣络方对脑缺血再灌注损伤(CIRI)大鼠SIRT1/PGC-1α信号通路的影响,初步探讨其对CIRI的保护机制。方法60只大鼠随机分为假手术组、模型组、活血荣络方组、EX-527组、EX-527+活血荣络方组和艾地苯醌组,采用线栓法建立CIRI大鼠模型。给药组于造模前36 h首次予相应药物灌胃(10 mL/kg),之后每24 h给药1次,共3次,EX-527组和活血荣络方+EX-527组于再灌注前20 min腹腔注射EX-527(5 mL/kg),其余各组在相同时间灌胃蒸馏水或腹腔注射生理盐水。再灌注24 h后对大鼠进行神经功能缺损评分,HE染色观察缺血皮质区损伤情况,试剂盒检测皮质区三磷酸腺苷(ATP)含量,免疫组化检测缺血皮质区沉默信息调节因子1(SIRT1)、过氧化物酶体增殖物激活受体γ辅激活因子1α(PGC-1α)蛋白表达,RT-qPCR检测缺血皮质区SIRT1、PGC-1αmRNA表达。结果与假手术组比较,模型组大鼠神经功能缺损评分升高,缺血皮质区神经元损伤严重,ATP含量减少,SIRT1、PGC-1α蛋白和mRNA表达降低(P<0.05)。与模型组比较,活血荣络方组和艾地苯醌组大鼠神经功能缺损评分降低,缺血皮质区神经元损伤改善,ATP含量增加,SIRT1、PGC1-α蛋白和mRNA表达升高(P<0.05);EX-527组大鼠神经功能缺损评分升高,缺血皮质区神经元损伤加重,ATP含量减少,SIRT1、PGC-1α蛋白和mRNA表达降低(P<0.05)。与活血荣络方组比较,活血荣络方+EX-527组大鼠神经功能缺损评分升高,缺血皮质区神经元坏死和凋亡增多,ATP含量减少,SIRT1、PGC-1α蛋白和mRNA表达降低(P<0.05)。结论活血荣络方可促进CIRI大鼠ATP生成,改善神经功能缺损症状,其机制可能与激活SIRT1/PGC-1α信号通路有关。

三七皂甙对脑组织血液供应和能量代谢的影响

目的研究三七皂甙（ＰＮＳ）对脑组织血液供应和能量代谢的影响。方法采用激光多普勒流量仪动态监测软脑膜微循环的变化以及反相高效液相法测定脑组织中高能磷酸化合物的含量。结果ＰＮＳ２５、５０ｍｇｋｇ－１ｉｖ明显扩张麻醉小鼠软脑膜微血管，增加局部血流量，加快血流速度；但对结扎双侧颈总动脉引起的脑血流量的减少没有作用。ＰＮＳ（５０、１００ｍｇｋｇ－１ｉｐ，×３ｄ）明显延缓缺血组织ＡＴＰ的分解，改善能量负荷。结论ＰＮＳ可通过增加组织血液供应改善能量代谢，从而保护脑组织。

异亚丙基莽草酸对大脑中动脉栓塞大鼠脑含水量和能量代谢的影响

目的 探究异亚丙基莽草酸 (ISA)减轻脑缺血损伤是否与改善能量代谢与减轻脑水肿有关。方法 采用三氯化铁致大脑中动脉栓塞模型 ,观察药物对脑组织含水量 ,能荷及Na+ ,K+ ATP酶的影响。结果 ISA 2 0 0mg·kg- 1可降低模型大鼠脑组织含水量 ,减轻脑水肿程度 ;ISA 2 0 0 ,10 0mg·kg- 1可明显增加模型大鼠脑组织能荷值 ;ISA 2 0 0 ,10 0 ,5 0mg·kg- 1对模型大鼠脑组织Na+ ,K+ ATP酶活力均有明显的提高作用。结论 ISA可能通过保护脑组织能量代谢和提高脑组织Na+ ,K+ ATP酶活力 ,减轻脑水肿程度 。

脑缺血时乳酸的生成、转运、代谢和利用

近年来对乳酸的研究表明,乳酸不仅是脑缺血/缺氧的代谢产物,同时也担负起在特殊情况下对神经元的供能作用。其主要在星形胶质细胞产生,又被神经元摄入胞内提供能量。此过程中单羧酸转运体(MCTs)充当着转运的角色。对乳酸的深入研究将有助于全面认识乳酸在脑缺血中的病理生理意义。

山莨菪碱在大鼠急性全脑缺血再灌注损伤中对一氧化氮内皮素-1及能量代谢的影响

目的 了解山莨菪碱在大鼠急性全脑缺血再灌注损伤中对一氧化氮（ＮＯ）、内皮素－１（ＥＴ－１）、ＡＴＰ及乳酸的影响，探讨山莨菪碱（Ａｎｉ）对脑缺血再灌注损伤的保护作用。方法采用Ｒｕｌｓｉｎｅｌｌｉ方法建立大鼠急性全脑缺血再灌注损伤模型。观察Ａｎｉ在再灌注期间对ＮＯ、ＥＴ－１及能量代谢的影响。结果Ａｎｉ可以抑制再灌注早期（１小时组）ＮＯ、ＥＴ－１及乳酸的过量产生，提高脑组织ＡＴＰ含量。结论 Ａｎｉ可以通过对ＮＯ、ＥＴ－１及能量代谢的影响对脑起保护作用。

丹酚酸B对脑缺血小鼠脑能荷和三磷酸腺苷酶活性的影响

目的:从脑缺血小鼠脑组织高能磷酸含量和三磷酸腺苷(ATP)酶活性探讨丹酚酸B对脑水肿的影响。方法:将NIH小鼠分为假手术组、缺血组、丹酚酸B治疗组和尼莫地平对照组,复制脑缺血模型。丹酚酸B组尾静脉注射丹酚酸B(22.5mg·kg^(-1));尼莫地平组尾静脉注射尼莫地平(0.03mg·kg^(-1));假手术组和缺血组尾静脉注射等量的生理盐水,30min后阻断双侧颈总动脉,观察缺血30min脑组织ATP、二磷酸腺苷(ADP)、一磷酸腺苷(AMP)和磷酸肌酸(PCr)的含量并计算能荷(EC)值;同时测定Na^+-K^+-ATP酶和Ca^(2+)-ATP酶的活性以及脑含水量。结果:与缺血组比较,丹酚酸B组脑组织ATP,ADP,PCr含量和能荷值显著升高(P<0.01);Na^+-K^+-ATP酶和Ca^(2+)-ATP酶活性显著增强(P<0.01);而脑含水量则明显降低(P<0.05)。结论:丹酚酸B减轻脑缺血小鼠脑含水量的作用机制,可能与其提高高能磷酸含量和增强ATP酶活性有关。

灯盏花素冰片注射液对实验性脑缺血的保护作用

目的:研究灯盏花素冰片注射液(SBI)对大鼠局灶性脑缺血和小鼠缺血缺氧性损伤的保护作用。方法:采用大鼠急性不完全性脑缺血实验,以大脑皮层含水量为指标初步筛选组方剂量。采用大鼠局灶性脑缺血再灌注损伤模型(MCAO)对模型大鼠进行神经症状和行为学评分,测定大脑皮层含水量,计算脑梗死区百分比,组织病理切片观察海马损伤程度,并测定脑组织内总抗氧化能力(T-AOC),丙二醛(MDA)和乳酸(LD)含量以及超氧化物歧化酶(SOD)、肌酸激酶(CK)、乳酸脱氢酶(LDH)、Na+-K+-ATP酶、Ca2+-Mg2+-ATP酶、总ATP酶(T-ATP)活力等生化指标;采用小鼠急性不完全性脑缺血实验和小鼠常压耐缺氧实验,测定其存活时及对耐缺氧能力的影响。结果:灯盏花素12mg/kg与冰片0.48mg/kg配伍对模型大鼠降低其脑含水量作用最为显著。与模型对照组比较,SBI组尾静脉注射能使动物的神经行为学评分,脑梗死率和脑含水量显著降低,能显著改善海马区神经元的形态,同时还能明显增强MCAO后大鼠脑组织内T-AOC能力,增加脑组织SOD,显著减少脑组织内MDA含量,同时对于脑组织内CK活力、LDH活力也有显著的增加作用,还能显著减少MCAO后大鼠脑组织LD含量,显著增加Na+-K+-ATP酶、Ca2+-Mg2+-ATP酶及T-ATP酶活力。结论:SBI高剂量组可以极显著延长急性不完全性脑缺血小鼠存活时间和耐缺氧能力。结论:SBI对实验性脑缺血具有保护作用。

异甘草素对小鼠脑缺血-再灌注损伤的保护作用

目的:研究异甘草素(ISL)对小鼠脑缺血再灌注损伤的保护作用。方法:采用反复阻断小鼠双侧颈总动脉造成脑缺血再灌注模型,断头法观察脑缺血再灌注小鼠耐缺氧能力;测定脑缺血再灌注小鼠全血粘度、红细胞压积和凝血时间;采用反相高效液相色谱法检测脑组织能量代谢水平。结果:ISL20,40mg·kg-1显著延长缺血再灌注小鼠断头后喘息时间(P<0.01),降低缺血再灌注小鼠全血粘度和红细胞压积(P<0.01),延长凝血时间(P<0.01);ISL(10～40mg·kg-1)剂量依赖性增加脑组织ATP含量、TAN水平和EC值,与模型组相比TAN水平和EC值分别增加了28%、42%、45%和41%、76%、90%,ATP含量依次恢复到假手术组的55%、75%、83%。结论:ISL可通过改善脑缺血再灌注小鼠异常血液流变学变化和提高脑组织能量代谢水平对脑缺血再灌注脑损伤起保护作用。

9602方对脑缺血再灌小鼠脑能量代谢的影响

目的 :观察 96 0 2方对脑缺血再灌小鼠脑能量代谢的影响。方法 :复制小鼠脑缺血再灌损伤模型 ,应用核磁共振 (NMR)技术和高效液相色谱 (HPLC)仪 ,检测小鼠脑能量代谢的变化。结果 :①NMR波谱显示 ,对照组和“96 0 2”组缺血 10min时 ,磷酸肌酸 (PCr)峰显著性下降的同时无机磷 (Pi)峰显著性上升 ,两组的PCr和Pi改变基本一致。不同之处在于 :再灌后对照组小鼠PCr继续缓慢下降 ,至再灌 10min时维持在低水平(5 5 5 0± 14 94) ;而“96 0 2”组再灌后 ,缺血时下降的PCr峰则开始回升 ,至再灌 10min时为 76 72± 13 37,两组相比有显著性差异 (P <0 0 5 )。再灌后对照组Pi峰则持续上升 ,至再灌 10min时为 10 9 76± 5 5 41;而“96 0 2”组在缺血时已升高的Pi峰则回降 ,至再灌 10min时为 79 99± 30 2 8,但两组相比无统计学意义。在整个低血压缺血再灌全过程中 ,ATP的 3个峰 (α、β、γ)均无明显变化。②HPLC仪检测显示 ,“96 0 2”组脑细胞能荷值 (0 2 884±0 0 5 5 2 )明显高于对照组 (0 110 4± 0 0 34 3) ,P <0 0 1。结论 :96 0 2方能改善脑缺血再灌所诱导的细胞和组织能量代谢的障碍。

蛛网膜下腔出血并迟发性脑血管痉挛大鼠脑能量代谢与脑血流量变化的实验研究

目的探讨实验性蛛网膜下腔出血导致的迟发性脑血管痉挛时脑能量代谢的变化规律,及其与皮质脑血流量(CBF)变化的相关性。方法50只雄性SD大鼠随机分为对照组和蛛网膜下腔出血第1、3、5、7天组,应用非抗凝自体动脉血枕大池两次注血法制备蛛网膜下腔出血大鼠模型;另5只大鼠用于颅内血液大体分布的观察。应用微透析仪监测各组大鼠脑组织间液中葡萄糖(Glu)、乳酸(Lac)及丙酮酸(Pyr)含量,并计算乳酸/丙酮酸比值;激光多普勒血流仪监测各组大鼠皮质脑血流量;于光学显微镜及透射电子显微镜下观察基底动脉血管壁的病理学改变。结果(1)5只用于观察颅内血液大体分布的大鼠中4只蛛网膜下腔出血模型鼠血液主要分布于基底池、脚间池及额叶底面,亚甲蓝与血液混合物主要沉积于颅底,同时在脑室及纵裂池亦可见血液分布;余1只注入人工脑脊液后未发生颅内出血及血液沉积。表明蛛网膜下腔出血动物模型制备成功。(2)与对照组相比,蛛网膜下腔出血组大鼠葡萄糖及丙酮酸含量明显降低(P<0.01);乳酸/丙酮酸比值明显升高(P<0.01),以第5、7天组升高最为明显(P<0.01);第5天组乳酸含量亦明显升高(P<0.01)。(3)激光多普勒监测显示,蛛网膜下腔出血组大鼠脑血流量减少率高于对照组(P<0.05),其中第5天组减少最为显著,与其他各亚组比较差异有统计学意义(P<0.05);脑血流量变化与乳酸/丙酮酸比值、乳酸水平的改变呈正相关(r=0.721,0.477;均P<0.01),与葡萄糖、丙酮酸水平的改变呈负相关(r=0.447,0.579;均P<0.01)。结论微透析指标与脑血流量变化具有一致性,可作为蛛网膜下腔出血脑组织间液生化指标的动态监测,用于预测迟发性脑血管痉挛的发生。

RP-HPLC法测定异甘草素对脑缺血再灌注小鼠脑能量代谢的影响

目的:建立一种测定组织样品中腺苷酸含量的 RP-HPLC 法,并观察异甘草素(isoliquiritigenin,ISL)对脑缺血再灌注小鼠能量代谢的改善作用。方法:采用反复阻断小鼠双侧颈总动脉造成脑缺血再灌注模型,采用 RP-HPLC 法,Hypersil C_(18)色谱柱(4.6mm×250mm,5μm),流动相为100mmol-L^(-1)磷酸二氢钾-磷酸氢二钾缓冲液(pH 6.0),流速为0.7mL·min^(-1),柱温为25℃,检测波长为254nm,测定脑组织 ATP 含量、腺苷酸池(TAN)水平和能量负荷值(EC)。结果:ATP、ADP、AMP 浓度分别在1.7-212.0μg·mL^(-1)(r=0.9999),1.6～208.0μg·mL^(-1)(r=0.9999),1.3～164.0μg·mL^(-1)(r=0.9999)范围内线性关系良好,平均回收率(n=6)分别为101.3%(RSD=1.0%),99.0%(RSD=1.4%),91.6%(RSD=1.6%);ISL(10,20,40mg·kg^(-1)·d^(-1))剂量依赖性提高脑组织中 ATP 含量、TAN 水平和 EC 值。结论:该分析方法简便、准确、可靠;ISL 对脑缺血再灌注小鼠脑能量代谢水平有明显的改善作用。

聪圣胶囊对鼠脑缺血损伤的保护及对脑血流与能量代谢的改善作用

目的 :研究聪圣胶囊对鼠脑缺血的保护作用。方法 :采用三氯化铁阻断大鼠一侧大脑中动脉、结扎大鼠双侧颈总动脉、高龄小鼠双侧颈总动脉反复缺血再灌注合并尾部放血的方法造成脑缺血模型 ,分别观察聪圣胶囊对神经症状、脑梗塞面积和脑含水量、脑血流量、脑组织能量代谢的影响。结果 :聪圣胶囊1、3、6g/kg显著降低脑梗塞面积和脑含水量 ,明显改善神经症状和脑血流量 ;3、6g/kg明显改善能量负荷。结论 :聪圣胶囊可通过增加脑组织血液供应 ,改善能量代谢 。

局灶性脑缺血时吡拉西坦对缺血半球能量代谢的影响

目的 观察吡拉西坦对局灶性脑缺血时缺血半球能量代谢的影响。方法 采用动脉腔内插线大鼠局灶性脑缺血模型及测定缺血 2 h/再灌注 2 h后缺血半球内乳酸 ( L A)水平、乳酸脱氢酶 ( L DH)和肌酸激酶 ( CK)活性的变化 ,HE染色结合图像分析测定缺血后 2 4h的梗死体积。结果 吡拉西坦 ( 2 0 0 mg/kg)可明显降低缺血半球内 L A水平 ,增加 L DH的活性 ,对 CK活性无明显影响 ,它可明显降低缺血 2 h/再灌注 2 2 h后的皮质、皮质下结构和半球的梗死体积。结论 吡拉西坦具有明确的抗局灶性脑缺血作用 ,它可明显改善缺血组织的能量代谢状态 ,增加 L DH的活性 ,减轻乳酸堆积及组织酸中毒 ,缩小梗死体积。

新生鼠缺氧缺血性脑损伤线粒体形态观察及定量研究

目的：观察及定量分析新生鼠缺氧缺血性脑损伤时脑选择性易损区海马CA1区锥体细胞线粒体形态改变。方法：制备新生鼠缺氧缺血性脑损伤模型，电镜观察、生物体视学测量线粒体形态。结果：缺氧缺血性脑损伤时线粒体肿胀，嵴溶解、断裂和消失。线粒体平均截面周长和平均截面积增加；比表面、嵴膜密度及嵴平均截线长减少。结论：缺氧缺血性脑损伤时线粒体形态发生显著变化。生物体视学定量分析准确反应了线粒体形态，尤其是功能区的改变。细胞内能量代谢障碍是脑损伤的重要环节。

超早期针刺百会、水沟对急性脑缺血大鼠能量代谢的影响

目的观察针刺抗脑缺血损伤的作用机理。方法采用微创开颅电凝法制作大鼠急性脑缺血模型,70只大鼠按随机数字表法分为7组:正常组、模型2 h组、模型24 h组,穴位2 h组、非穴位2 h组、穴位24 h组、非穴位24 h组,每组10只。针刺百会、水沟(频率120次/min,捻转泻法1 min,5 min/次,留针30 min)。非穴位针刺分别在百会、水沟左侧旁开5 mm处进针,手法同穴位针刺。穴位24 h组及非穴位24 h组在24 h再针刺1次。采用高效液相检测各组缺血脑组织ATP、ADP、AMP含量,比色法检测缺血脑组织Na+-K+ATP酶、Ca2+ATP酶含量。结果造模成功后,脑组织ATP、ADP、Na+-K+ATP酶和Ca2+ATP酶含量在大鼠脑缺血后2 h后即明显下降,AMP含量明显升高,与正常组比较,差异明显(P<0.01),且缺血24 h后尤为显著,与2 h比较有极显著差异(P<0.01);穴位组大鼠ATP、ADP、Na+-K+ATP酶和Ca2+ATP酶含量较模型组及非穴位组比较明显升高,AMP含量下降,有极显著差异(P<0.01),并且穴位2 h组优于24 h组,有显著差异(P<0.05);相同时间点非穴位组与模型组比较,无显著差异(P>0.05)。结论穴位针刺可以有效改善脑缺血大鼠脑组织能量代谢,发挥抗脑缺血损伤的作用,进一步为临床针刺抗脑缺血提供实验依据。

心脑舒通胶囊对大鼠急性脑缺血损伤保护作用的研究

目的:观察与分析心脑舒通胶囊对急性脑缺血损伤过程中能量代谢障碍、自由基损伤及炎性因子表达等关键环节的影响,探讨缺血保护作用机制。方法:健康雄性W istar大鼠60只,随机分为正常组、假手术组、模型组、心脑舒通组、西药组,共5组,每组12只。心脑舒通组灌胃给予心脑舒通混悬液,西药组给予阿司匹林加尼莫地平混悬液,正常组、假手术组及模型组均给予等量蒸馏水。术前3 d开始连续灌胃给药10 mL.kg-1,每天1次,至术后3 d共7 d。采用同种系微栓子体外注入法制备大鼠多发性脑梗死模型,缺血72 h后断头取脑,常规HE染色光镜下观察患侧海马CA1区及皮层病理学改变,免疫组化SABC法染色光镜下观察肿瘤坏死因子-α(TNF-α)及白细胞介素-1β(IL-1β)在海马CA1区及皮层的表达,采用比色法测定海马组织中Na+-K+-ATP酶活性、乳酸脱氢酶(LDH)活力、丙二醛(MDA)含量及超氧化物歧化酶(SOD)活性。结果:心脑舒通胶囊能明显减轻急性多梗大鼠海马CA1区的病理形态改变,不同程度的减少海马CA1区及皮层神经细胞内TNF-α及IL-1β表达,显著提高海马组织中ATP酶活性(P<0.01)及LDH酶活力(P<0.01),能显著提高海马组织中SOD活性(P<0.01)、降低MDA含量(P<0.01)。结论:心脑舒通胶囊具有一定的缺血损伤保护作用,其机制与改善缺血组织的能量代谢障碍和自由基损伤,抑制炎性因子过表达,多环节阻抑和调节缺血级联反应有关。