大鼠实验性脾虚证胰腺组织化学研究

用成年雄性Ｗｉｓｔａｒ大鼠３０只,分为（１）正常对照组,喂饲自来水。（２）脾虚组,用苦降破气中药和饮食失节法致成脾虚模型。（３）自然恢复组,动物致虚后,喂饲自来水。（４）中药治疗组。取四组动物胰腺进行ＲＮＡ,琥珀酸脱氢酶（ＳＤＨ）,乳酸脱氢酶（ＬＤＨ）,三磷酸腺苷酶（ＡＴＰａｓｅ）,葡萄糖－６－磷酸酶（Ｇ－６－Ｐａｓｅ）和硫胺素焦磷酸酶（ＴＰＰａｓｅ）组织化学反应和观察,并对ＳＤＨ,ＬＤＨ,ＲＮＡ进行了显微分光光度计定量测定。本研究结果表明,脾虚组胰腺泡细胞的ＲＮＡ,ＳＤＨ,ＡＴＰａｓｅ,Ｇ－６－Ｐａｓｅ,ＴＰＰａｓｅ含量和活性都低于对照组,而ＬＤＨ活性高于对照组。治疗组与自然恢复组相比,治疗组胰腺泡细胞以上指标接近对照组。定量测定与定性的结果一致。本研究表明,脾虚证时胰腺泡细胞上述几种酶活性和ＲＮＡ明显下降,可能在脾虚证发病中起主要作用,中药治疗有显著改善     

维拉帕米对骨骼肌缺血再灌注损伤保护作用的酶组织化学研究

实验选用大鼠骨骼肌缺血再灌注模型, 观察骨骼肌缺血及再灌注后酶组织化学和超微结构的变化, 并观察维拉帕米的保护作用。结果显示： 骨骼肌缺血６ｈ 时, 骨骼肌细胞ＳＤＨ、ＣＣＯ、Ｃａ２＋ －ＡＴＰａｓｅ活性呈下降趋势, 而ＬＤＨ 活性则有所增强。再灌注１２ｈ 时, 骨骼肌细胞ＳＤＨ、ＣＣＯ、Ｃａ２＋ －ＡＴＰａｓｅ 活性进一步明显下降,同时ＬＤＨ 活性亦下降明显,而应用维拉帕米能在一定程度上保护上述酶的活性。与此同时,骨骼肌超微结构的改变与其酶活性的变化相一致。因此, 本实验提示： 骨骼肌缺血再灌注可损害其能量代谢酶的活性, 而维拉帕米则有较强的保护作用。     

丹参对鼠缺血后残肝酶组织化学的影响

研究观察鼠缺血后肝再生时酶组织化学的变化。结果表明缺血后肝再生时肝细胞ＳＤＨ、Ｇ６Ｐａｓｅ和Ｍｇ２＋ＡＴＰａｓｅ活性及ＰＡＳ反应均在不同程度上低于对照组,而ＬＤＨ、ＡＣＰ活性较对照组有不同程度升高。丹参组酶活性及ＰＡＳ反应基本处于缺血组与对照之间,说明丹参对保护某些酶活性,促进肝再生有一定的作用     

铝对大白鼠海马所致损害的实验研究

为探讨铝对海马的损害,用３０只健康大白鼠,体重２００～３００克,分为Ａ组（正常对照）１０只,Ｂ组（低铝组饲料中加５００ｍｇ／ｋｇＡｌＣｌ３）１０只,Ｃ组（高铝组饲料中加２５００ｍｇ／ｋｇ／ＡｌＣｌ３）１０只。分别饲养１２个月后,分组断头取脑海马,按组织化学及酶组织化学常规做“铝定性”反应,琥珀酸脱氢酶（ＳＤＨ）、乳酸脱氢酶（ＬＤＨ）、硫胺素焦磷酸酶（ＴＰＰａｓｅ）、镁激活三磷酸腺苷酶（Ｍｇ２＋－ＡＴＰａｓｅ）、胆碱酯酶（ＣｈＥ）测定。按电镜常规做超微结构的观察。结果提示：Ｃ组,海马ＳＤＨ、Ｍｇ２＋－ＡＴＰａｓｅ、ＣｈＥ活性明显减弱（由强阳性（）下降为阳性（＋）,铝反应增强（在海马的神经细胞内强阳性（）;同时海马神经元内线粒体嵴断裂、肿胀、空泡样变,神经纤维髓鞘变性（如不规则卷曲、松散）。说明铝对海马的功能与结构均有损害     

骤冷与饥饿对小鼠肝脏影响的实验研究

为探讨饥饿及饥饿与骤冷对动物肝脏的影响,本实验用健康昆明种小鼠２５只,随机分成正常组５只,饥饿组１０只,饥饿后再予冷刺激组１０只（下称骤冷组）。采用组织化学及酶组织化学方法观察糖原（ＰＡＳ反应）、ＳＤＨ（琥珀酸脱氢酶）,ＬＤＨ（乳酸脱氢酶）,ＣｈＥ（胆碱酯酶）、Ｍｇ２＋－ＡＴＰａｓｅ（镁激活三磷酸腺苷酶）,ＡＣＰ（酸性磷酸酶）。结果提示：饥饿时肝脏ＰＡＳ反应,ＳＤＨ,Ｍｇ２＋－ＡＴＰａｓｅ、ＣｈＥ活性显著下降,而ＡＣＰ活性明显增强;饥饿后骤冷时ＰＡＳ（反应）、ＳＤＨ、ＣｈＥ更显著下降,而ＡＣＰ及Ｍｇ２＋－ＡＴＰａｓｅ活性反而增强。     

甲状腺素对人体缺血/再灌注心肌能量代谢酶影响的定性定量研究

用酶组织化学方法检测了缺血／再灌注心肌ＬＤＨ、ＣＣＯ、ＳＤＨ、Ｃａ－ＡＴＰａｓｅ 活性; 分组研究甲状腺素（ＴＨ） 对围术期心肌能量代谢的影响。结果： 用药组ＳＤＨ、ＣＣＯ、Ｃａ－ＡＴＰａｓｅ 的活性比对照组增高, 两组ＬＤＨ无差异; 对照组１ 例死亡, 术前、术中、术后ＳＤＨ、ＣＣＯ、ＬＤＨ、Ｃａ－ＡＴＰａｓｅ 的活性显著降低。结论： 术前补充ＴＨ, 可提高心肌的有氧代谢, 能量合成, 促进心功能; 如果术前能了解心肌各酶活性, 将对手术适应症的选择提供帮助     

小儿先天性巨结肠症酶组织化学变化的研究

小儿先天性巨结肠症（ＨＤ）之病因不甚清楚。国内外学者对其狭窄段各种神经递质的研究较多。而针对细胞能量代谢有关酶类变化研究很少。故本实验对２０例ＨＤ病儿的狭窄段,正常段全层组织进行酶组织化学方法的观察,根据酶反应的强弱分组对比观察。结果发现：（１）ＡＴＰａｓｅ（腺苷三磷酸酶）,ＳＤＨ（琥珀酸脱氢酶）,Ｃｈｅ（胆碱脂酶）,狭窄段酶活性显著高于正常段。（２）ＭＡＯ（单胺氧化酶）活性狭窄段明显低于正常设。（３）狭窄段粘膜下及肌间神经丛,和神经节细胞平均个数明显少于正常段。     

亚硒酸钠对四氯化碳损伤草鱼肝原代细胞与肝组织的保护作用

不同浓度四氯化碳（ＣＣｌ４）对草鱼肝原代细胞的损伤实验中,ＣＣｌ４浓度为１０μｌ／ｍｌ可引起细胞血清中丙氨酸氨基转移酶（ＡＬＴ）、天门冬氨酸氨基转移酶（ＡＳＴ）、乳酸脱氢酶（ＬＤＨ）逸出量与细胞破损率显著增高,培养液中添加亚硒酸钠（Ｎａ２ＳｅＯ３）０．２μｇ／ｍｌ,则可降低ＡＬＴ、ＡＳＴ、ＬＤＨ的逸出量,减轻细胞破损程度。Ｎａ２ＳｅＯ３保护实验中,Ｎａ２ＳｅＯ２＋ＣＣｌ４组预先腹腔注射（ｉｐ）０．１ｍｇ／ｋｇ．ｂｗ连续三日,末次ｉｐＣＣｌ４混合液１ｍｌ／ｋｇ．ｂｗ,２４ｈ内肝组织超氧物歧化酶（ＳＯＤ）相对活性比ＣＣｌ４组提高达９１．５％,第七日仍提高达５４．５％,与对照组的水平基本接近;血清中丙氨酸氨基转氨酶（ＡＬＴ）水平逐渐降低。本实验还观察到Ｎａ２ＳｅＯ３可引起肝脂质过氧化物显著降低,肝微粒体蛋白含量与细胞色素Ｐ—４５０活性升高;组织切片观察显示肝组织损伤程度减轻,７２ｈ后细胞核增多。表明Ｎａ２ＳｅＯ３可提高草鱼肝清除自由基能力,增强肝脏解毒功能。     

肝缺血再灌流损伤和复方丹参保护作用的实验研究

观察大鼠肝脏缺血后再灌流时酶组织化学及超微结构的变化,同时观察复方丹参的保护作用。结果显示：肝缺血２小时后再灌流３小时,肝ＳＤＨ、Ｍｇ２＋－ＡＴＰａｓｅ、Ｇ－６－Ｐａｓｅ的活性明显降低,ＬＤＨ、ＡＣＰ的活性明显增强。超微结构变化表现为肝细胞内线粒体肿胀、嵴断裂、空泡样变,粗面内质网颗粒明显减少,肝窦扩大、窦壁内皮细胞肿胀,胆小管扩张,微绒毛减少、断裂。复方丹参对肝细胞的缺血再灌流损伤有明显的保护作用     

三唑酮对绿豆幼苗叶片衰老的延缓作用

三唑酮处理可提高离体绿豆（ＰｈａｓｅｏｌｕｓｒａｄｉａｔｕｓＬ．）幼苗叶片叶绿素和蛋白质含量。叶片衰老过程中超氧物歧化酶（ＳＯＤ）、过氧化物酶（ＰＯＤ）、过氧化氢酶（ＣＡＴ）和抗坏血酸过氧化物酶（ＡｓＡＰＯＤ）活性及抗坏血酸（ＡｓＡ）和还原型谷胱甘肽（ＧＳＨ）含量降低。２０ｍｇ／Ｌ三唑酮可提高ＰＯＤ、ＡｓＡＰＯＤ活性和ＡｓＡ、ＧＳＨ含量,对ＳＯＤ、ＣＡＴ活性无影响。丙二醛（ＭＤＡ）含量在叶片衰老过程中提高,并与ＰＯＤ、ＡｓＡＰＯＤ活性和ＡｓＡ、ＧＳＨ含量呈显著负相关,三唑酮可降低ＭＤＡ含量。表明三唑酮有提高植物对膜脂过氧化作用的保护能力,延缓叶片的衰老作用。     

RCS大鼠和Wistar大鼠视网膜酸性磷酸酶活性的动态观察

本实验观察了不同年龄组ＲＣＳ大鼠和Ｗｉｓｔａｒ大鼠视网膜中酸性磷酸酶的动态变化及其与ＲＰＥ细胞消化功能的关系。运用偶氮偶联法显示１２ｄ、２１ｄ、２ｍ的ＲＣＳ大鼠和７ｄ、２ｍ的Ｗｉｓｔａｒ大鼠视网膜中的酸性磷酸酶;通过图像分析仪测定ＲＰＥ细胞层和光感受器外节部分的酸性磷酸酶含量,并进行统计学分析。结果：酸性磷酸酶阳性反应呈暗红色,主要位于ＲＰＥ细胞层,视网膜外核层、内核层,节细胞层亦有少量阳性反应颗粒。２ｍ的ＲＣＳ大鼠视细胞内、外节的酸性磷酸酶含量则明显高于其它组（Ｐ＜０．０１）,其余结构的酸性酶各组间无显著性差异（Ｐ＞０．０５）。结论：ＲＣＳ大鼠和Ｗｉｓｔａｒ大鼠的视网膜色素上皮细胞可能具有相同的消化功能。     

老龄大鼠内侧隔核BDNF含量的变化及人参皂甙的保护作用

目的探讨老龄大鼠内侧隔核BDNF含量的变化情况及人参皂甙的保护作用,为人参皂甙的临床应用提供实验依据.方法选用Wistar大鼠40只,分成青年组(3个月)、老龄组(26个月)、给药组(自17个月始给予人参皂甙至26个月),采用免疫组织化学方法检测大鼠内侧隔核BDNF含量的变化情况.结果老龄组内侧隔核BDNF阳性反应物的平均灰度值与青年组相比显著增高(P<0.01),而给药组与老龄组相比其平均灰度值显著降低(P<0.01).结论老龄大鼠内侧隔核BDNF含量明显低于青年组,给药组BDNF含量较老龄组明显增加.说明人参皂甙具有保护BDNF,防止神经元退行性改变的作用.     

人参皂甙对老龄大鼠海马结构BDNF及Trk B蛋白表达的影响

目的探讨老龄大鼠海马结构BDNF及Trk B蛋白表达的老龄性变化,同时对比观察人参皂甙对其改变的影响。方法雌性Wistar大鼠39只,分为青年组、老龄组、给药组(第17个月始饲以人参皂甙至27月龄)。采用免疫组化及Western blot方法对海马结构BDNF反应产物及Trk B蛋白进行定性、定量分析。结果老龄组CA3、CA1区BDNF含量分别较青年组下降13.3%、10.4%(P<0.05);其给药组和老龄组相比变化不大(P>0.05)。齿状回从青年到老年变化不明显(P>0.05),但给药组比老龄组增加16.7%(P<0.01)。老龄组海马结构Trk B蛋白表达较青年组下调了99.7%;给药组较老龄组上调了78.5%(P<0.01)。结论老龄组海马结构BDNF及Trk B蛋白表达较青年组明显降低,而人参皂甙可明显上调齿状回BDNF含量和海马结构Trk B蛋白的表达。     

人参皂甙对老龄大鼠Meynert核TrkB mRNA表达的影响

目的探讨人参皂甙对老龄大鼠Meynert核(NBM)神经元TrkB mRNA表达的影响.方法选40只雌性Wistar大鼠随机分为3组:青年组(3-5月龄)、老龄组(24-27月龄)及给药组(24-27月龄).给药组大鼠自18月始饲以人参皂甙至24-27月龄.对各组NBM神经元进行原位杂交组织化学法染色,采用图象分析法测定各组平均灰度值并作t检验.结果老龄组大鼠NBM神经元TrkB mRNA表达显著低于青年组,而给药组较老龄组大鼠表达增多.结论人参皂甙对NBM神经元TrkB mRNA表达有促进作用,本结果为人参皂甙抗脑衰老及AD的临床治疗提供了形态学依据.     

Quantitative succinate dehydrogenase histochemistry in the hippocampus of aged rats

Quantitative histochemical methods (microphotometric kinetic and end-point measurements, and morphometric analyses of reactive areas) were used to investigate the levels of succinate dehydrogenase (SDH) in the hippocampus of young adult (3-6 months old) and aged male rats (24-27 months old). Methodological studies concerning the demonstration of SDH activity, which were performed using hippocampi of young animals, revealed a linear relationship between the reaction time and the amount of reaction product for up to 20 min; kinetic (continuous) and end-point measurements provided the same results. In a number of experiments, it was established that an incubation medium consisting of 100 mM succinate, 10 mM sodium azide, 3 mM nitro blue tetrazolium chloride, 0.25 mM phenazine methosulfate, and 7.5% polyvinylalcohol in 0.05 M Hepes buffer (final pH 7.5) was optimal for quantitative SDH histochemistry in the hippocampus. Comparative quantitative investigations of SDH activity in rat hippocampi showed that, in most regions and layers of the hippocampus of both young and aged rats, the levels of SDH activity increased along the rostrocaudal axis of the hippocampus, i.e., higher levels were present in the caudal than in the rostral pole. In both groups, the highest SDH levels were observed in the molecular layer of the cornu ammonis (CA)-1, the CA-3, and the fascia dentata (middle and outer thirds), most of which are termination fields of the excitatory perforant path arising from the regio ento-rhinalis. Furthermore, in almost all of the investigated layers, the older animals exhibited lower SDH levels than young animals. These differences were statistically significant in the molecular layer of the fascia dentata and in most layers of the CA-3. The lower SDH levels in aged animals are discussed in relation to the reduced capacity for energy metabolism in the aging brain.     

Quantitative succinate dehydrogenase histochemistry in the hippocampus of aged rats

Summary Quantitative histochemical methods (microphotometric kinetic and end-point measurements, and morphometric analyses of reactive areas) were used to investigate the levels of succinate dehydrogenase (SDH) in the hippocampus of young adult (3–6 months old) and aged male rats (24–27 months old). Methodological studies concerning the demonstration of SDH activity, which were performed using hippocampi of young animals, revealed a linear relationship between the reaction time and the amount of reaction product for up to 20 min; kinetic (continuous) and end-point measurements provided the same results. In a number of experiments, it was established that an incubation medium consisting of 100 mM succinate, 10 mM sodium azide, 3 mM nitro blue tetrazolium chloride, 0.25 mM phenazine methosulfate, and 7.5% polyvinylalcohol in 0.05M Hepes buffer (final pH 7.5) was optimal for quantitative SDH histochemistry in the hippocampus. Comparative quantitative investigations of SDH activity in rat hippocampi showed that, in most regions and layers of the hippocampus of both young and aged rats, the levels of SDH activity increased along the rostrocaudal axis of the hippocampus, i.e., higher levels were present in the caudal than in the rostral pole. In both groups, the highest SDH levels were observed in the molecular layer of the cornu ammonis (CA)-1 the CA-3, and the fascia dentata (middle and outer thirds), most of which are termination fields of the excitatory perforant path arising from the regio entorhinalis. Furthermore, in almost all of the investigated layers, the older animals exhibited lower SDH levels than young animals. These differences were statistically significant in the molecular layer of the fascia dentata and in most layers of the CA-3. The lower SDH levels in aged animals are discussed in relation to the reduced capacity for energy metabolism in the aging brain.Dedicated to Professor Dr. T.H. Schiebler on the occasion of his 65th birthdaySupported by the Deutsche Forschungsmeinschaft (Ku 541/2-1)     

Senescence-associated beta-galactosidase activity expression in aging hippocampal neurons

To investigate the activity of senescence-associated beta-galactosidase (SA-beta-GAL) in the hippocampus of aging rats. Hippocampi of 6-, 18-, and 24-month-old rats were observed by histochemical staining for SA-beta-GAL and cytochemical staining for SA-beta-GAL in cultured hippocampal neurons. The activity of SA-beta-GAL doubled in hippocampal pyramidal cells of the CA3 region in rats between 6 and 18 months (14.57 ± 2.74% vs. 31.66 ± 14.12% SA-beta-GAL-positive, respectively), and reached 50.76 ± 14.41% positive at 24 months. The activity of SA-beta-GAL also increased as a function of time upon prolonged culture of cultured hippocampal neurons with 95% of cells being SA-beta-GAL-positive at 20 days in vitro. Interestingly, no SA-beta-GAL-positive cells were found in neurons of the hippocampal dentate gyrus, a neurogenic region of the brain, at any age examined. SA-beta-GAL can be used as a senescence biomarker in determining senescent neurons in hippocampal pyramidal cells of the CA3 region in advanced aging.     

异丙肾上腺素导致大鼠心肌损伤的酶组织化学及超微结构观察

健康成年Ｗｉｓｔａｒ大鼠５０只,雌雄各半,体重２１０±２０ｇ。正常对照组１０只,实验组４０只,每只大鼠皮下注射盐酸异丙肾上腺素（ＩＳＰ）５．０ｍｇ／ｋｇ,在注射后４、１２、２４、４８小时,分别处死１０只大鼠,取出心脏,进行（１）ＨＥ染色,（２）组化染色,按Ｐｅａｒｓｏｎ法显示ＳＤＨ,用物镜测微网计算每组ＳＤＨ活性产物的缺失面积;（３）超微结构观察,按立体学图像分析原理,用《多功能医学图像处理系统》微机计算各组电镜片中线粒体体密度值。本实验的观察表明：心肌细胞ＳＤＨ在注射ＩＳＰ后显示出不同的阶段性变化;ＳＤＨ的活性产物缺失部位与ＨＥ染色的心肌坏死部位一致;心肌ＳＤＨ的活性变化与线粒体的损伤同时出现,并与线粒体的破坏程度密切相关。     

胎脑提取液对衰老小鼠海马神经元酶活性影响的实验研究

目的 观察胎脑提取液对衰老小鼠海马神经元酸性磷酸酶（ACP）和琥珀酸脱氢酶（SDH）活性的影响,探讨胎脑提取液的抗衰老作用。方法 选用健康昆明种小白鼠30只,随机分为3组;采用D-半乳糖制备亚急性衰老模型;酶组织化学方法结合显微图像分析。观察各组小鼠海马神经元ACP和SDH的活性。结果 衰老模型组与正常对照组相比,小鼠海马神经元ACP活性明显升高,SDH活性明显降低;给药组与衰老模型组相比,小鼠海马神经元ACP活性明显降低。SDH活性明显升高。结论 胎脑提取液可以延缓海马神经元的衰老进程,具有一定的抗衰老作用。     

党参对冠心病心绞痛患者的血液细胞及对小鼠心肌作用的定量细胞化学观察

本文应用细胞化学和定量细胞化学技术,对冠心病心气虚病人服用党参前后外周血细胞中的糖原及琥珀酸脱氢酶（ＳＤＨ）的含量和活性,以及党参对小鼠心肌中糖原、ＳＤＨ、乳酸脱氢酶（ＬＤＨ）的影响进行了分析研究。结果发现,党参可明显提高冠心病人外周血细胞中糖原和ＳＤＨ的含量和活性;同时动物实验亦证实,灌服党参液的小鼠心肌中糖原、ＳＤＨ、ＬＤＨ的含量和活性亦明显高于生理盐水组。这说明党参可改善冠心病人的心肌代谢,对冠心病的治疗是十分有利的。