益气养阴消癥通络中药对高糖联合AngⅡ培养的大鼠系膜细胞p38MAPK信号通路的影响

目的：探讨益气养阴消癥通络中药对高糖联合血管紧张素Ⅱ培养的大鼠肾小球系膜细胞（MCs）p38MAPK信号通路的作用。方法：原代培养MCs,第5代时,予高糖、高糖联合血管紧张素Ⅱ刺激,并同时进行药物血清干预,48h后ELISA法检测细胞上清ColⅣ蛋白含量,Western blot检测MCs p-p38MAPK、p38MAPK、p-STAT3、STAT3蛋白表达水平。结果：中药血清可显著减少细胞上清ColⅣ蛋白含量（P〈0.01）,且能明显抑制p38MAPK、STAT3蛋白磷酸化水平（P〈0.01）。结论：益气养阴消癥通络中药可能是通过下调p38MAPK信号通路表达,减少细胞外基质合成,从而发挥对肾脏的保护作用。     

NGF促进骨折愈合的适宜浓度初步研究

目的通过研究局部应用不同浓度NGF对骨折愈合的影响,进一步探讨NGF促进骨折愈合的适宜浓度。方法成年雄性SD大鼠75只,体重(220.0±2.5)g,随机分成A、B、C、D、E 5组(n=15)。实验动物建立右胫骨中上段骨折模型,A、B、C、D组分别于骨折断端局部注射含0.006 48×10-2、0.032 40×10-2、0.162 00×10-2、0.810 00×10-2μg/g NGF的生理盐水0.3 mL,每天1次,连续注射7 d;E组注射等量生理盐水。各组分别于术后2、4、6周各处死5只大鼠,并于骨折部位取材行大体、X线片观察、生化检测及组织学观察。处死大鼠前取大鼠眼球球后动、静脉血液行血清ALP活性测定。结果术后各时间点,大体观察表明A、B、C、D组局部新生硬组织大小、硬度、骨折断端骨痂生长逐渐递增,D组明显优于A、B、C、E组。X线片示A、B、C、D组骨折线逐渐模糊,钙化面积逐渐增大,D组骨痂钙化明显快于A、B、C组和E组。组织学观察示D组骨小梁质量及成熟度明显优于A、B、C组和E组。D组术后4、6周成骨区灰度值、骨痂钙含量,2、4周骨痂组织湿重及2周血清ALP含量均显著高于A、B、C组和E组,差异有统计学意义(P<0.05)。术后2、4周,骨组织形态计量分析显示各组新生骨骨小梁表面成骨细胞指数、骨小梁面积百分比和骨小梁宽度随时间延长而减小;同一时间点A、B、C、D组依次增大,均大于E组,且差异均有统计学意义(P<0.05)。结论局部应用NGF对大鼠骨折愈合有促进作用,高浓度的NGF(0.810 00×10-2μg/g)对骨折愈合有显著促进作用。     

刺五加多糖对H2O2诱导的大鼠海马神经元凋亡的保护作用

目的研究刺五加多糖（ASPS）对H2O2诱导的海马神经元凋亡的影响及其机制。方法采用H2O2诱导大鼠海马神经元凋亡。采用末端脱氧核苷酸转移酶介导的dUTP原位切口末端标记法检测细胞凋亡率、免疫组化法检测caspase-3蛋白的表达、逆转录PCR法检测caspase-3 mRNA的表达。结果H2O2作用后,海马神经元凋亡率、caspase-3蛋白和mRNA表达水平均显著增高（P〈0.05）;给予ASPS干预后,均显著下降（P〈0.05）;而且,随ASPS剂量增加,作用效果显著增强（P〈0.05）。结论ASPS具有抑制氧化应激损伤诱导神经细胞凋亡作用,其机制与下调caspase-3 mRNA的表达有关。     

乌司他丁对大鼠胰性脑病保护作用的实验研究

目的 :探讨乌司他丁对大鼠急性出血坏死性胰腺炎时对脑组织损害的保护作用 ,为胰性脑病的治疗提供实验依据。方法 :将 90只SD大鼠随机分为正常对照组 ,急性出血坏死性胰腺炎诱导组和乌司他丁处理组 ,经胰胆管逆行注射 5%牛磺胆酸钠建立大鼠ANP模型 ,12h后处死动物 ,测定血清TNFα水平 ,并检测脑组织含水量、脑微血管内白细胞聚集及附壁现象。结果 :经乌司他丁处理后 ,血清中乌司他丁水平、脑组织含水量、脑组织MDA含量、脑微血管内白细胞聚集及附壁现象显著降低(P <0 .0 1)。结论 :乌司他丁可以减轻急性出血坏死性胰腺炎时脑损害的发生和发展     

8 Hz 130 dB次声作用对大鼠肾上腺超微结构的影响

目的：探讨次声作用对大鼠肾上腺细胞损伤特点及意义。方法：将雄性SD大鼠42只随机分为对照组和1、7、14、21、28和35d实验组，实验组在8HZ 130 dB次声环境下每日暴露1次，每次2h；取肾上腺组织固定，透射电镜下观察肾上腺细胞超微结构变化。结果：不同环境和天数次声作用后肾上腺皮质细胞可见细胞膜破裂，发生变性坏死。其损伤随作用次数增多其损伤逐渐加重，28、35d组损伤程度有所恢复。各实验组肾上腺髓质细胞超微结构变化不明显。结论：次声作用可对肾上腺细胞造成不同程度损伤，28d后肾上腺对次声的损伤作用存在着一定适应现象。     

立体定向毁损杏仁核对大鼠脑内单胺递质影响的实验研究

目的 :探讨立体定向毁损正常大鼠杏仁核对相关脑区单胺递质含量的影响。方法 :将 75只雄性Sprague -Dawley大鼠随机分为对照组、假手术组和手术组 ,每组 2 5只。测定立体定向毁损正常大鼠杏仁核后额叶、间脑和脑干多巴胺 (DA)、五羟色胺 (5 -HT)和去甲肾上腺素 (NE)含量的变化。DA、5 -HT测定采用荧光分光光度法 ;NE测定采用放射免疫法。结果 :毁损后大鼠额叶、间脑和脑干DA显著降低于对照组 (P <0 .0 1) ;NE显著降低于对照组 (P <0 .0 5 ,P <0 .0 1,P <0 .0 1)。毁损后大鼠额叶、间脑和脑干 5 -HT均显著高于对照组 (P <0 .0 5 ,P <0 .0 5 ,P <0 .0 1)。结论 :立体定向毁损正常大鼠杏仁核后能使相关脑区单胺递质含量发生改变。     

伊文思蓝、zo-1评估血脑屏障损伤的应用研究

目的 :寻找新的简捷、快速、精确、灵敏的评估血脑屏障 (BBB)损伤的指标和方法。方法 :应用脑缺血再灌注损伤模型 ,造成大鼠BBB的损伤 ,采用形态学和化学定量两种新方法观察BBB对伊文思蓝的通透性 ,应用免疫组织化学法检测脑组织中ZO - 1蛋白的表达水平的变化 ,以反映BBB的损伤程度。结果 :①脑局限性缺血再灌注损伤后脑内ZO - 1蛋白水平迅速降低 ,至再灌注后 2 4h达最低水平 ,至再灌注 7d时恢复到正常水平 ;②两种方法检测的伊文思蓝最大通透率均出现在再灌注损伤第 2 4h ,其变化趋势与脑组织中ZO - 1蛋白水平的变化趋势相一致。结论 :BBB对伊文思蓝的通透性和脑组织中ZO - 1蛋白水平的变化均可用于反映BBB损伤的程度 ,形态学观察BBB对伊文思蓝通透率的方法可灵敏地反映BBB的损伤程度     

A change of P-selectin immunoreactivity in rat brain after transient and permanent middle cerebral artery occlusion

Abstract

Although the role of an adhesion molecule such as P-selectin may be important in the pathogenesis of stroke, temporal, spacial, and cellular profiles of the expression ofsuch a protein has not been fully studied in the case ofthe middle cerebral artery (MCA) occlusion and reperfusion in rat brain. Change in expression of immunoreactive P-selectin was examined in rat brain after transient MCA occlusion (MCAO) in comparison to that of permanent occlusion with an anti-P-selectin monoclonal antibody. Western blot analyses were performed to ensure the selective detection of immunoreactive P-selectin protein with the monoclonal antibody using brain homogenates before and after MCAO. Temporal, spacial, and cellular changes of P-selectin expressions were evaluated with rat brain sections at 2, 8 h, 1 and 3 days of permanent MCAO, and at 2, 8 h, 1, 3 and 7 days of reperfusion after 1 h of transient MCAO. Western blot showed a single band with a molecular weight of 140 kOa for both cases with permanent occlusion and reperfusion. P-selectin immunoreactivity was not normally present in rat brain sections. However, it was expressed mainly in the post-capillary venules of the cerebral cortex and caudate in the MCA territory with a peak at 2-8 h after permanent occlusion and at 8 h to 1 day after the reperfusion. The expression was diminished by 1 day ofpermanent occlusion and 3 days of reperfusion. The maximum staining in the case of permanent MCAO was stronger than the case with reperfusion. However, spacial distribution of the staining was similar in the cerebral cortex and caudate between the cases with permanent or transient MCAO. These results suggest a different temporal but similar spacial and cellular expression of P-selectin immunoreactivity between permanent occlusion and reperfusion of MCA in rat brain. [Neural Res 1998; 20: 463–469]     

大鼠液压脑损伤后皮层微血管改变与脑水肿的关系

目的探讨大鼠液压脑损伤后皮层微血管损伤情况及其与伤后脑水肿的关系。方法成年SD大鼠30只,随机分为正常组（n=6）、假手术组（n：6）、损伤组（n=18）,其中损伤组分为伤后6h、24h、72h三亚组,每亚组6只。利用液压冲击法建立大鼠颅脑损伤模型,显微镜下观察直接损伤侧和非直接损伤侧皮层微血管损伤隋况,CD34标记血管内皮细胞评价血管密度改变,干湿重法检测脑组织含水量的变化。结果大鼠皮层微血管损伤后6h可见血管支行迂曲、扩张、充血,伤后24h可见少量血栓形成,损伤后72h可见有较多血栓形成。损伤组CD34阳性细胞数明显低于假手术组和对照组（P〈0．05）,而脑组织含水量明显高于假手术组和对照组（P〈0．05）,而后两组无统计学差异（P〉O．05）。损伤组直接损伤侧皮层微血管损伤较非直接损伤组严重,而且伤后24h较伤后6、72h严重。结论颅脑损伤后脑微血管损伤为全脑性血管损伤,这可能是伤后脑水肿形成的机制之一。     

Isolation and characterization of beta 1-adrenergic receptors from adult, rat cardiac myocytes

The beta-receptors were isolated from rat cardiac myocytes and characterized. Isolated myocytes were prepared from adult rat hearts and characterized for viability. Membrane proteins were solubilized from myocytes with 1% Triton X-102. The solubilized membrane proteins were fractionated by DEAE-Sephacel ion exchange column chromatography. Two major protein peaks were obtained. The second protein peak sample was found to contain beta-receptors to which 125I-15-(4'-azido-3'-iodobenzyl)-carazolol (125I-ABC) was specifically bound. This sample was labeled covalently with 125I-ABC by UV irradiation. The radiolabeled sample was applied to a Sepharose CL-6B gel column. Two radiolabeled protein peaks, one with a molecular weight of approximately 570,000 and the other with a molecular weight of approximately 95,000 were found. When the 570,000-dalton complex was subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) under reducing conditions, it was dissociated into a component with a molecular weight of 66,000. The 95,000-dalton complex was dissociated into a 58,000-dalton component upon SDS-PAGE under reducing conditions. An excess amount of isoproterenol and propranolol decreased photolabeling of the beta-receptors with 125I-ABC by 60% and 40%, respectively.