The effect of radix Salviae Miltiorrhizae (RSM) on substance P in cerebral ischemia--animal experiment.

The levels of substance P (SP) in rat brains were assayed in 64 rats. Bilateral common carotid artery ligation was done in 49 rats. Half an hour before ligation, 25 rats were given 10 g/kg of RSM; 24 rats were given the same volume of normal saline as controls. Sham operation was done in 15 rats. Half an hour and 3 hours after cerebral ischemia, the rats were quickly decapitated. SP concentration was assayed in the cerebral cortex, caudate nucleus and brain stem. In saline-treated animals, the SP level of caudate nucleus at 3-hour group was significantly decreased as compared with the 0.5-hour group and sham-operated group respectively. No significances were found among RSM-treated groups and sham-operated groups. The SP levels were shown: brain stem greater than caudate nucleus greater than cerebral cortex. The preliminary results suggest that SP may be involved in the pathophysiologic procedures of cerebral ischemia and RSM may attenuate the dysfunction of SP during cerebral ischemia.     

Effect of radix Salviae miltiorrhizae on extracellular adenosine and evaluation of its protective efficacy in ischemic reperfusion rat--microdialysis, HPLC and histopathologic studies.

The effects of Radix Salviae Miltiorrhizae (RSM) on extracellular adenosine (Ade) and its metabolites, i.e. inosine, hypoxanthine and xanthine, were studied with microdialysis and HPLC techniques during cerebral ischemia-reperfusion induced by 4-vessel occlusion in rat brain. Histological examination of hippocampus was performed 6 h after reperfusion. ECF (extracellular fluid) adenosine and its metabolites were compared between the controls (n = 6) and RSM-treated rats (n = 6). Basal level of Ade and its metabolites release were not greatly affected by pretreatment with RSM, and no significant difference as compared with the sham-operated (n = 6). Ade and its metabolites were dramatically increased after ischemia, and decreased near basal-level and its metabolites remained high at the end of reperfusion. In the RSM-treated animals, the tendency of changes of Ade and its metabolites was just the same as in the controls, but the magnitudes of changes were significantly lower at some different time points. In sham-operated animals, no changes were observed at different time points both during ischemia (30 min.) and reperfusion (60 min.). Histopathological findings demonstrated that RSM pretreatment results in better histologic preservation of the pyramidal cells in the postischemic reperfusion CA1 sector both qualitatively and quantitatively. These results indicated that RSM protects against cerebral ischemia reperfusion injury.     

ET-1 gene expression of rat brain during ischemia and reperfusion and the protective effect of radix Salviae miltiorrhizae.

Endothelin-1 (ET-1) gene expression of rat brain during ischemia and reperfusion as well as the effect of Radix Salviae Miltiorrhizae (RSM) were studied with in situ hybridization. It was found that ET-1 gene expression of cerebral cortex and caudate-putamen was markedly increased both in 24 hours of ischemia and 24 hours of reperfusion groups (P < 0.01, P < 0.05). In RSM-treated rats, although the ET-1 gene expressions of ischemia and reperfusion sides were also increased as compared with contralateral cortex and caudate-putamen, they were significantly lower in RSM-treated rats than those of controls (P < 0.05, P < 0.01 respectively). The present study indicated that RSM can partly inhibit ET-1 gene expression of cerebral cortex and caudate-putamen during ischemia and reperfusion. This may be one of the protective mechanisms of RSM on cerebral ischemia and reperfusion.     

The effect of radix Salviae miltiorrhizae on vasoactive intestinal peptide in cerebral ischemia: an animal experiment

The levels of vasoactive intestinal peptide (VIP) in three regions of rat brain were assayed in 62 rats. Bilateral common carotid artery ligation was done in 50 rats. Half an hour before ligation 26 rats were given 10 g/kg of Radix Salviae Miltiorrhizae (RSM); 24 rats were given same volume of normal saline as controls. A sham operation was done in 12 rats. Half an hour (n = 30) and 3 hours (n = 32) after operation, the rats were quickly decapitated. VIP levels were assayed in cerebral cortex, hippocampus and caudate nucleus. In salin-treated animals, VIP levels of cerebral cortex and caudate nucleus at 3 hour group were significantly decreased compared with the sham-operated group. No significant difference was found between RSM-treated and sham-operated groups. The preliminary results suggest that VIP may be involved in the pathophysiological procedures of cerebral ischemia and RSM may attenuate the dysfunction of VIP during cerebral ischemia.     

Effect of radix Salviae miltiorrhizae on the gene expression of nitric oxide synthase in ischemic rat brains.

The effect of Radix Salviae Miltiorrhizae (RSM) on the gene expression of nitric oxide synthase (NOS) in rat brains during ischemia was studied with in situ hybridization and the results were analyzed with IBAS 2000 Image Analysis System. It was found that NOS gene expression of cerebral cortex and caudate-putamen was markedly increased in 24 hours in ischemia group (P < 0.01). In RSM-treated rats, although the NOS gene expression of ischemic side was also increased as compared with contralateral cortex and caudate-putamen, it was significantly lower in RSM-treated rats than those of the controls (P < 0.05, P < 0.01). The present study indicates that RSM can partly inhibit NOS gene expression of cerebral cortex and caudate-putamen during ischemia. This may be one of the protective mechanisms of RSM on cerebral ischemia.     

丹参对脑缺血后脑组织中血管活性肠肽（VIP）的影响

本实验以结扎大白鼠双侧颈总动脉为脑缺血模型,测定了62只大白鼠脑组织中的血管活性肠肽(VIP)。结果发现生理盐水组脑缺血后3小时皮层和尾状核VJP含量显著低于假手术组,海马中VIP含量亦有下降趋势;丹参组与假手术组比较各脑区VIP含量无显著差异。实验结果提示VIP参与了脑缺血的病理生理过程,丹参可能具有缓解脑缺血后VIP紊乱的作用。     

丹参对肝脏缺血再灌注后病理变化影响的实验研究

观察丹参对肝脏热缺血和冷缺血再灌注后病理变化的影响。方法 :将家兔 12只制成肝脏热缺血模型 ,电镜下观察热缺血再灌注后丹参对肝脏超微结构变化的影响 ;对 2 0头猪施行辅助性肝移植术 ,光镜下观察移植肝在冷缺血再灌注后丹参对肝脏病理变化的影响。结果 :兔热缺血再灌注后 ,在肝细胞超微结构观察中 ,发现丹参可明显减轻线粒体和粗面内质网肿胀 ;猪冷缺血再灌注后 ,在肝脏光镜观察中 ,发现丹参可明显减轻肝细胞浊肿、脂变和汇管区炎症。结论 :丹参可明显减轻肝脏热缺血和冷缺血再灌注后肝脏病理损害。     

缺血-再灌注大鼠胃黏膜毛细血管超微结构的改变

目的 探讨缺血-再灌注大鼠胃黏膜毛细血管超微结构的改变.方法 SD雄性大鼠随机分为正常对照组（CON）、单纯缺血组（ISC）、缺血再灌注1 h组（I/R1）、24 h组（I/R2）和72 h组（I/R3）,用透射电镜观察大鼠胃缺血-再灌注损伤过程中胃黏膜毛细血管超微结构的变化.结果 单纯缺血组,毛细血管内皮细胞肿胀,可见核内异染色质向核周聚集;缺血再灌注1 h时,内皮细胞肿胀明显,可见内皮细胞核固缩,管腔闭塞;再灌注24 h后,毛细血管内皮细胞损伤减轻;再灌注72 h后基本恢复正常.结论 缺血-再灌注大鼠胃黏膜毛细血管超微结构有变化,内皮细胞呈现细胞凋亡的特征,提示胃黏膜毛细血管形态的改变与胃缺血-再灌注损伤的发生发展密切相关.     

局灶性脑缺血早期大鼠尾壳核生长抑素mRNA表达的变化

目的观察早期脑缺血对大鼠尾壳核内生长抑素(SOM)mRNA表达的影响。方法采用原位分子杂交方法,观察大脑中动脉阻塞所致脑缺血1 h和6 h后大鼠尾壳核SOM mRNA表达的变化。结果脑缺血早期缺血侧大鼠尾壳核内SOM mRNA表达明显下降(P<0.01)。结论早期脑缺血对大鼠尾壳核SOM mRNA表达有明显的影响,提示SOM mRNA阳性神经元可能对脑缺血耐受性较差。     

Effects of transient forebrain ischemia and radix Salviae miltiorrhizae (RSM) on extracellular levels of monoamine neurotransmitters and metabolites in the gerbil striatum--an in vivo microdialysis study.

The aim of this study was to investigate the effect of 30 min forebrain ischemia, followed by 120 min reperfusion on extracellular fluid (ECF) levels of dopamine (DA), norepinephrine (NE), serotonin (5-HT) and their metabolites, homovanillic acid (HVA) and 5-hydroxyindoleacetic acid (5-HIAA) in the striatum of gerbils, so as to obtain further information on the mechanism of Radix Salviae Miltiorrhizae (RSM)-induced neuroprotection. Microdialysis was used to sample the extracellular space. Dialysate was measured by high performance liquid chromatography with electrochemical detector (HPLC-ED). ECF DA, NE levels increased from basal levels by 282, 227 and 221 folds, by 9.14, 8.51 and 8.25 folds, respectively for the three ischemic duration (0-10; 11-20; 21-30 min). ECF DA, NE, 5-HT levels in the RSM-treated group were significantly decreased as compared with those in the control group during ischemia (P < 0.01). The results suggested that monoamine neurotransmitters were involved in ischemic neuron damage directly or indirectly; and that RSM plays a protective role during cerebral ischemia by attenuating the dysfunctions of monoamine neurotransmitters.     

Protective effect of radix Salviae miltiorrhizae on apoptosis of neurons during focal cerebral ischemia and reperfusion injury.

We have found that Radix Salviae Miltiorrhizae (RSM) plays a protective role in ischemic brain injury, which attracted us to investigate the effect of RSM on apoptosis of neurons during cerebral ischemia and reperfusion. The apoptotic cells in ischemic brains at different reperfusion intervals were tested with the method of TdT-mediated dUTP-DIG nick end labeling (TUNEL), and the effect of RSM on the apoptosis of neurons was studied in left middle cerebral artery (LMCA) occlusion in rat models (n = 18). The results showed that few scattered apoptotic cells were observed in right cerebral hemisphere after LMCA occlusion and reperfusion, and that a lot of apoptotic cells were found in left ischemic cerebral cortex and caudoputamen at 12 h reperfusion, and they reached peak at 24-48 h reperfusion. However, in rats pretreated with RSM, the number of apoptotic cells in left cortex and caudoputamen reduced significantly and the neuronal damage was much milder at 24 h reperfusion as compared with those of saline-treated rats. From this study, we conclude that administration of RSM can reduce the apoptotic of neurons induced by cerebral ischemia and reperfusion and afford significant cerebroprotection in the model of focal cerebral ischemia and reperfusion.     

兔眼冲击伤后早期视网膜超微结构改变

目的:通过对兔眼冲击波损伤后早期视网膜超微结构改变的动态观察,探讨眼冲击波损伤后视网膜结构改变及演变过程。方法:用激波管制成兔眼冲击波损伤动物模型,分别于伤后不同时点处死动物,摘取全眼球,取视飞纹下方后极部全层眼球壁,在透射电镜下观察其超微结构改变。结果:冲击波造成了一系列的视网膜结构改变。首先表现为视网膜视细胞结构损伤,如视细胞的外节出现微丝微管的断裂肿胀和结构紊乱、视盘肿胀和崩解、视细胞核肿胀碎裂;继而出现视网膜水平细胞和节细胞的反应性肿胀,线粒体等膜性细胞器明显肿胀,细胞间骨架破坏;随后视网膜内网状层肿胀,节细胞膜性细胞器明显肿胀,细胞间骨架破坏消失,细胞间结构异常、溶解消失。结论:眼冲击波损伤后视网膜视细胞的早期超微结构改变可能是冲击波的直接致伤效应所致,节细胞的结构紊乱可能是继发性改变,均是不可逆性视功能损害的病理改变基础。     

雨蛙素诱发的大鼠急性胰腺炎胰腺毛细血管透射电镜观察

目的：了解实验性急性胰腺炎时胰腺毛细血管超微结构的改变并探讨其与急性胰腺炎的关系,方法：采用皮下注射雨蛙素建立大鼠急性水肿性胰腺炎模型,用透射电镜观察胰腺毛细血管形态细胞的变化。结果：毛细血管腔内表面不规则,毛细血管内皮细胞核增大、线粒体肿胀、内皮细胞向腔内的突起增加,内皮细胞胞质内蚕饮小泡及“窗孔”数量均明显增加。结论：急性胰腺炎早期即存在胰腺毛细血管内皮细胞的损伤,推测其与胰腺微血管通透性增高有关,并导致胰腺局部区域血液灌流不同,最终引起组织缺血,加速病变的发展。     

银杏内酯B对局灶性脑缺血时星形胶质细胞GFAP表达的影响

目的 观察血小板活化因子(PAF)受体拮抗剂银杏内酯B(GB)对局灶性脑缺血时星形胶质细胞胶质纤维酸性蛋白(GFAP)表达的影响,并探讨其作用机制.方法 建立光化学诱导树局灶性脑缺血模型,用HE染色和电镜技术观察缺血后不同时间脑组织星形胶质细胞的形态学改变;用免疫组织化学法观察脑缺血后4 h、24 h、72 h及给予GB后24 h半暗区及对侧皮层星形胶质细胞GFAP的表达,并测定其平均灰度值.结果 光镜下可见,HE染色显示树局灶性脑缺血后,随缺血时间延长,星形胶质细胞形态有不同程度改变;电镜观察显示缺血24 h时星形胶质细胞明显肿胀.免疫组织化学染色可见,半暗区星形胶质细胞GFAP表达在4 h时没有明显改变, 24 h时增加 (P＜0.01),72 h时仍维持在较高水平(P＜0.01);对侧皮层星形胶质细胞GFAP表达于72 h时开始增加(P＜0.05).缺血后6 h舌下静脉注射GB至缺血24 h时,星形胶质细胞GFAP表达少于缺血组(P＜0.05),但仍较假手术组高.结论 脑缺血后星形胶质细胞GFAP表达增强,GB可通过减少星形胶质细胞GFAP的表达而起脑保护作用.     

心肌缺血-再灌注损伤的超微结构变化

目的：观察犬急性心肌缺血－再灌注损伤的早期超微结构改变,探讨急性心肌缺血－再灌注损伤所致急死的形态学基础。方法：复制犬急性心肌缺血－再灌注损伤动物模型,在透射电镜下观察心肌纤维、线粒体、内质网和微血管结构变化。结果：缺血30min再灌注120min,心肌细胞线粒体轻度肿胀,基质内可见絮状致密体;缺血60～90min再灌注120min,心肌细胞线粒体嵴断裂,基质内出现絮状致密体和颗粒状致密体。微血管内皮细胞肿胀,血管腔狭窄。结论：再灌注可使缺血30min心肌超微结构得到部分恢复,加重缺血60～90min心肌超微结构改变。     

实验性高粘血症对脑微血管内皮细胞的影响

①目的探讨高粘血症对家兔脑微血管内皮细胞结构的影响。②方法通过每天静脉推注５ｍｌ／ｋｇ体重６％高分子右旋糖酐，使实验家兔血液呈高粘滞状态，应用光镜及透射电镜观察大脑皮层（顶叶）微血管内皮细胞形态。③结果家兔顶叶脑膜及皮层内微血管扩张充血，红细胞聚集，毛细血管内皮肿胀，突起增多增大，质膜小泡显著增多，基膜增厚，并常见肥大细胞附着于基膜外。④结论持续性高粘血症可使兔脑微血管内皮细胞的结构与功能发生改变。     

针刺对脑缺血再灌注大鼠皮质缺血半暗带细胞超微结构的影响

目的：观察脑缺血再灌注后,针刺对脑缺血再灌注大鼠皮质缺血半暗带细胞超微结构的影响。方法采用改良 Longa 氏线栓法制作局灶性脑缺血再灌注大鼠模型,用多聚甲醛灌注,戊而醛固定,醋酸铀和枸橼酸铅双重染色后,HU-12A 型电镜下观察大鼠皮质缺血半暗带细胞超微结构的变化。结果模型组6 h 神经元轻度损伤,染色质边集凝固,血管周围水肿,水肿渗出明显,毛细血管管腔狭窄,脑组织散在水肿,脑细胞破坏;针刺组5 d 后神经元结构正常,血管通畅,毛细血管内皮细胞无损伤,脑组织无水肿、渗出、无坏死区域,新生毛细血管增多。结论针刺能够减轻脑水肿,较少神经元细胞的损伤,促进新生毛细血管的增生,从而达到保护脑组织的目的。     

心肌缺血再灌注对大鼠心肌和血管内皮细胞超微结构的影响

目的:观察心肌缺血再灌注对心肌和血管内皮细胞超微结构的影响.方法:电镜下观察心肌缺血再灌注模型大鼠心肌缺血部位超微结构改变.结果:心肌细胞明显肿胀;肌膜破坏;染色质凝聚、边集;肌原纤维松弛,形成异常收缩带,肌丝断裂、溶解,形成大量肌溶灶;线粒体肿胀,嵴断裂、缺失、空泡化;三联体松解;糖原明显减少.血管内皮细胞肿胀,染色质边集,线粒体增大,吞饮小泡明显减少,应力纤维分散,梗死灶内血管内皮细胞变性、断裂和分离.结论:缺血再灌注后对心肌和血管内皮细胞超微结构造成明显的损伤,但血管内皮细胞较心肌纤维损伤轻.     

内源性NGF在缺血性老年大鼠部分脑区及小脑中的表达

目的 探讨缺血性老年大鼠部分脑区及小脑与神经生长因子（neve growth factor,NGF)之间的关系。方法 用夹闭两侧颈总动脉法建立不完全性脑缺血动物模型，将SD大鼠随机分成正常对照组（A组）、假手术组（B组）、缺血30min，再灌注6h（C组）、12h（D组）、24h（E组）、48h（F组）、7d(G组）和14d（H组），用免疫组织化学SLAB染色法检测部分脑区神经元及小脑NGF的表达。在透射电镜下观察各组神经元超微结构变化。结果 除A，B组外，各组顶叶皮质神经元均有NGF表达，其中E组和G组中量表达，各组海马均有少量表达，各组额叶少量表达，小脑均没有表达；再灌注超过48h组海马神经元和小脑蒲肯野细胞损伤较重，顶叶皮质神经元轻度水肿。结论 老年大鼠在受到缺血性脑损伤时，内源性NGF对脑组织具有一定的保护作用。     

大脑完全缺血后形态学和代谢功能的改变Ⅱ

结扎蒙古沙土鼠颈总动脉,造成脑缺血模型,松解后,观察存活期14天、21天、28天的实验动物的大脑形态学和代谢功能的改变。结果显示大脑皮质、尾状核、海马和豆状核壳等部位对缺血损害最为敏感。存活4天～21天的动物对2-DG的摄取明显回升并缓慢恢复,血流恢复后28天的脑组织,其神经元结构出现不同阶段的恢复。